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1. Bozepinib, a novel small antitumor agent, induces PKR-mediated apoptosis and synergizes with IFNα triggering apoptosis, autophagy and senescence

Author

Marchal JA, Carrasco E, Ramírez A, Jiménez G, Olmedo C, Peran M, Agil A, Conejo-García A, Cruz-López O, Campos JM, and García MA

Subjects
Therapeutics. Pharmacology, RM1-950
Abstract

Juan Antonio Marchal,1,2 Esther Carrasco,1 Alberto Ramirez,1,3 Gema Jiménez,1,2 Carmen Olmedo,4 Macarena Peran,1,3 Ahmad Agil,5 Ana Conejo-García,6 Olga Cruz-López,6 Joaquin María Campos,6 María Ángel García4,7 1Biopathology and Regenerative Medicine Institute, Centre for Biomedical Research, 2Department of Human Anatomy and Embryology, Faculty of Medicine, University of Granada, Granada, 3Department of Health Sciences, University of Jaén, Jaén, 4Experimental Surgery Research Unit, Virgen de las Nieves University Hospital, Granada, 5Department of Pharmacology and Neurosciences Institute, Faculty of Medicine, 6Department of Pharmaceutical and Organic Chemistry, Faculty of Pharmacy, University of Granada, Granada, 7Department of Oncology, Virgen de las Nieves University Hospital, Granada, Spain Abstract: Bozepinib [(RS)-2,6-dichloro-9-[1-(p-nitrobenzenesulfonyl)-1,2,3,5-tetrahydro-4,1- benzoxazepin-3-yl]-9H-purine] is a potent antitumor compound that is able to induce apoptosis in breast cancer cells. In the present study, we show that bozepinib also has antitumor activity in colon cancer cells, showing 50% inhibitory concentration (IC50) values lower than those described for breast cancer cells and suggesting great potential of this synthetic drug in the treatment of cancer. We identified that the double-stranded RNA-dependent protein kinase (PKR) is a target of bozepinib, being upregulated and activated by the drug. However, p53 was not affected by bozepinib, and was not necessary for induction of apoptosis in either breast or colon cancer cells. In addition, the efficacy of bozepinib was improved when combined with the interferon-alpha (IFNα) cytokine, which enhanced bozepinib-induced apoptosis with involvement of protein kinase PKR. Moreover, we report here, for the first time, that in combined therapy, IFNα induces a clear process of autophagosome formation, and prior treatment with chloroquine, an autophagy inhibitor, is able to significantly reduce IFNα/bozepinib-induced cell death. Finally, we observed that a minor population of caspase 3-deficient MCF-7 cells persisted during long-term treatment with lower doses of bozepinib and the bozepinib/IFNα combination. Curiously, this population showed ß-galactosidase activity and a percentage of cells arrested in S phase, that was more evident in cells treated with the bozepinib/IFNα combination than in cells treated with bozepinib or IFNα alone. Considering the resistance of some cancer cells to conventional chemotherapy, combinations enhancing the diversity of the cell death outcome might succeed in delivering more effective and less toxic chemotherapy. Keywords: seven-member heterocycles, purines, IC50, interferon cytokine, cell death, breast and colon cancer cells

Published
2013

3. Abstract P6-05-01: Interactions between adipocytes and breast cancer cells stimulate cytokine production and drive Src/SOX2/miR-302b mediated malignant progression

Author

Picon-Ruiz, M, primary, Pan, C, additional, Drewes-Elger, K, additional, Jang, K, additional, Besser, A, additional, Zaho, D, additional, Morata-Tarifa, C, additional, Kim, M, additional, Ince, TA, additional, Azzam, D, additional, Wander, S, additional, Cote, RJ, additional, Guy, HA, additional, El-Ashry, D, additional, Torne-Poyatos, P, additional, Marchal, JA, additional, and Slingerland, JM, additional

Published
2018
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6. Antitumoural Properties of Benzannelated Seven -Membered 5-fluorouracil Derivatives and Related Open Analogues. Molecular Markers for Apoptosis and Cell Cycle Dysregulation

Author

ESPINOSA A, MARCHAL JA, ARANEGA A, GALLO MA, AIELLO, Stefania, CAMPOS J., ESPINOSA A, MARCHAL JA, ARANEGA A, GALLO MA, AIELLO S, and CAMPOS J

Subjects
Antitumour drugs, Breast cancer, Cell cycle, Programmed cell death, Benzodioxepins
Abstract

Attention is increasingly being focussed on the cell cycle and apoptosis as potential targets for therapeutic intervention in cancer. We prepared a series of bioisosteric benzannelated seven-membered 5-FU O,N-acetals to test them against the MCF-7 human breast cancer cell line. Benzo-fused seven-membered O,O-acetals or their acyclic analogues led to the expected 5-FU O,N-acetals (or aminals), in addition to six- and 14-membered aminal structures and acyclic compounds. All the cyclic aminals provoked a G0/G1-phase cell cycle arrest, whereas Ftorafur, a known prodrug of 5-FU, and 1-[2-(2-hydroxymethyl-4-nitrophenoxy)-1-methoxyethyl]-5-fluorouracil (11) induced an S-phase cell cycle arrest. Although breast cancer is most often treated with conventional cytotoxic agents it has proved difficult to induce apoptosis in breast cancer cells, but improved clinical responses may be obtained by identifying therapies that are particularly effective in activating apoptosis. 1-(2,3-Dihydrobenzoxepin-2-yl)-5-fluorouracil (5) may be particularly useful in stimulating apoptosis in breast cancer.

Published
2005

7. Design of a low frequency, impulsive sound simulator in an existing house for sonic boom perceptual studies

Author

Cretagne Léo, Garcia A. Carlos, Leconte Roman, Ollivier François, Marchal Jacques, Marmel Frédéric, Fritz Claudia, and Coulouvrat François

Subjects
sonic boom, simulator, Acoustics in engineering. Acoustical engineering, TA365-367, Acoustics. Sound, QC221-246
Abstract

The renewal of civil supersonic aviation is partly conditioned by the establishment of an international regulation on sonic boom level. Human perception of booms from future aircraft creating sound disturbances of lower level than past ones can currently be evaluated only through boom simulators in laboratory setups with predicted signatures from numerical simulations. To reach sufficient ecological validity, it is necessary that perception studies take place in an environment as familiar as possible to participants. With this in view, a simulator has been designed to reproduce sonic booms of low amplitude with the highest possible fidelity and control, while adapting to an existing house. The article presents the challenges and design solutions chosen to reach this objective. A double optimisation of the input signal, successively in the frequency and in the time domain, is described. Observed performances are presented for different boom exposures and in various rooms of the house.

Published
2023
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8. Release of alpha-actin into serum after skeletal muscle damage.

Author

Martínez-Amat A, Boulaiz H, Prados J, Marchal JA, Puche PP, Caba O, Rodríguez-Serrano F, and Aránega A

Abstract

OBJECTIVE: The skeletal muscle protein alpha-actin was investigated in the serum of subjects with severe skeletal muscle damage to assess its utility as a reliable and predictive marker of muscle damage. METHODS: Serum samples were obtained from 33 healthy controls and 33 patients with severe skeletal muscle damage, defined by a total creatine kinase value of >500 IU/l (Rosalki method). Troponin I, troponin T, and myoglobin concentrations were determined by immunoassay and alpha-actin concentrations by Western blot and densitometry. RESULTS: The mean serum concentration of alpha-actin in controls and patients with skeletal muscle damage was 600.9 and 1968.51 ng/ml, respectively, a statistically significant difference. Sera of patients with muscle damage showed higher levels of alpha-actin than of troponin or myoglobin. No significant difference in troponin I levels was observed between the groups. CONCLUSIONS: According to these results, alpha-actin was the most significant skeletal muscle damage marker analysed and may be an ideal candidate for the identification of all types of myofibre injury, including sports injuries. Our findings support the use of alpha-actin as a marker alongside other currently used biological proteins. [ABSTRACT FROM AUTHOR]

Published
2005
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9. Cognitive development in patients with Mucopolysaccharidosis type III (Sanfilippo syndrome)

Author

Colland Vivian, Grootenhuis Martha, Marchal Jan, Valstar Marlies J, and Wijburg Frits A

Subjects
Medicine
Abstract

Abstract Background Mucopolysaccharidosis type III (MPS III, Sanfilippo syndrome) is a lysosomal storage disorder caused by a deficiency of one of the enzymes involved in the degradation of heparan sulfate. MPS III is characterized by progressive mental deterioration resulting in severe dementia. A number of potentially disease-modifying therapies are studied. As preservation of cognitive function is the ultimate goal of treatment, assessment of cognitive development will be essential in order to evaluate treatment efficacy. However, no large scale studies on cognitive levels in MPS III patients, using formal psychometric tests, have been reported. Methods We aimed to assess cognitive development in all 73 living patients with MPS III in the Netherlands. Results Cognitive development could be assessed in 69 patients. In 39 of them developmental level was estimated > 3 months and formal psychometric testing was attempted. A remarkable variation in the intellectual disability was detected. Conclusions Despite special challenges encountered, testing failed in only three patients. The observed broad variation in intellectual disability, should be taken into account when designing therapeutic trials.

Published
2011
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10. Silver Nanoparticles from Annona muricata Peel and Leaf Extracts as a Potential Potent, Biocompatible and Low Cost Antitumor Tool

Author

María G. González-Pedroza, Houria Boulaiz, René García-Contreras, Yaiza Jiménez-Martínez, Eduardo Martínez-Martínez, Saúl A Navarro-Marchal, Liliana Argueta-Figueroa, Juan A. Marchal, Raúl A. Morales-Luckie, [González-Pedroza,MG] Department of Biotechnology, Faculty of Sciences, Autonomous University of the State of Mexico (UAEMex), Toluca, Mexico. [Argueta-Figueroa,L] Conacyt Chairs, Faculty of Dentistry, Benito Juárez Autonomous University of Oaxaca, Oaxaca, Mexico. [García-Contreras,R] Interdisciplinary Research Laboratory, Nanostructures and Biomaterials, National School of Higher Studies (ENES) León Unit, National Autonomous University of Mexico (UNAM), Leon, Mexico. [Jiménez-Martínez,Y, Navarro-Marchal,SA, Marchal,JA, Boulaiz,H] Regenerative Institute of Biopathology and Medicine (IBIMER), University of Granada, Granada, Spain. [Martínez-Martínez,E] Laboratory of Cell Communication and Extracellular Vesicles, National Institute of Genomic Medicine (INMEGEN), Mexico City, Mexico. [Navarro-Marchal,SA] Department of Applied Physics, Faculty of Sciences, University of Granada, Granada, Spain. [Navarro-Marchal,SA, Boulaiz,H] Biosanitary Institute of Granada (ibs. GRANADA), SAS—Universidad de Granada, Granada, Spain. [Navarro-Marchal,SA, Boulaiz,H] Research Unit 'Modeling Nature' (MNat), University of Granada, Granada, Spain. [Marchal,JA, Boulaiz,H] Department of Human Anatomy and Embryology, University of Granada, Granada, Spain. [Morales-Luckie,RA] Joint Center for Research in Sustainable Chemistry UAEMex—UNAM (CCIQS), Autonomous University of the State of Mexico (UAEMex), Toluca, Mexico., and This research was supported by Fundacion Empresa Universidad de Granada (Project PR/18/001), Fundación Mutua Madrileña (Project FMM-AP16683-2017), Consejería de Salud Junta de Andalucía (PI-0089-2017) and Instituto de Salud Carlos III (RTI2018-101309-B-C22)

Subjects
antiproliferative activity, Frutas, Plata, Biocompatibility, Acetogenins, Chemicals and Drugs::Chemical Actions and Uses::Specialty Uses of Chemicals::Laboratory Chemicals::Indicators and Reagents::Reducing Agents [Medical Subject Headings], General Chemical Engineering, Terapéutica, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Therapeutics [Medical Subject Headings], Anatomy::Plant Structures::Plant Components, Aerial::Fruit [Medical Subject Headings], Context (language use), 02 engineering and technology, Antiproliferative activity, Silver nanoparticle, Annona, 03 medical and health sciences, 0302 clinical medicine, Therapeutic index, Anatomy::Cells::Cells, Cultured::Cell Line [Medical Subject Headings], Annona muricata fruit peel, acetogenins, General Materials Science, Nanopartículas del metal, QD1-999, Annona muricata, Organisms::Eukaryota::Plants::Viridiplantae::Streptophyta::Embryophyta::Angiosperms::Annonaceae::Annona [Medical Subject Headings], Diseases::Neoplasms::Neoplasms by Histologic Type::Nevi and Melanomas::Melanoma [Medical Subject Headings], chemistry.chemical_classification, Tecnología química ecológica, biology, Traditional medicine, green synthesis of silver nanoparticles, Chemistry, Technology and Food and Beverages::Technology, Industry, and Agriculture::Manufactured Materials::Nanostructures::Nanoparticles::Metal Nanoparticles [Medical Subject Headings], Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Chemistry Techniques, Analytical::Surface Plasmon Resonance [Medical Subject Headings], Green synthesis of silver nanoparticles, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Chemistry Techniques, Analytical::Spectrum Analysis::Spectrophotometry::Spectrophotometry, Infrared::Spectroscopy, Fourier Transform Infrared [Medical Subject Headings], Diseases::Neoplasms [Medical Subject Headings], 021001 nanoscience & nanotechnology, biology.organism_classification, Biocompatible material, Neoplasias, 030220 oncology & carcinogenesis, Cancer cell, Chemicals and Drugs::Inorganic Chemicals::Metals::Metals, Heavy::Silver [Medical Subject Headings], 0210 nano-technology, Lactone
Abstract

Cancer is one of the most prevalent diseases in the world and requires new therapies for its treatment. In this context, the biosynthesis of silver nanoparticles (AgNPs) has been developed to treat different types of tumors. The Annona muricata plant is known for having anticancer activity. Its main compounds present in the leaves, stems and skin, allowing for its use as reducing agents. In this manuscript, AgNPs with leaf extract (AgNPs-LE) and fruit peel extract (AgNPs-PE) of A. muricata were biosynthesized obtaining an average nanoparticle diameter sizes smaller than 50 nm, being 19.63 ± 3.7 nm and 16.56 ± 4.1 nm, and with a surface plasmonic resonance (SPR) at 447 and 448 nm, respectively. The lactone functional group present in the LE and PE extracts was identified by the FTIR technique. The behavior and antiproliferation activity of AgNPs-LE and AgNPs-PE were evaluated in breast, colon and melanoma cancer cell lines. Our results showed that Annona muricata fruit peel, which is a waste product, has an antitumor effect more potent than leaf extract. This difference is maintained with AgNPs where the destruction of cancer cells was, for the first time, achieved using concentrations that do not exceed 3 μg/mL with a better therapeutic index in the different tumor strains. In conclusion, we present a low-cost one-step experimental setup to generate AgNPs-PE whose in-vitro biocompatibility and powerful therapeutic effect make it a very attractive tool worth exploiting.

Published
2021

11. Evolution of Metastasis Study Models toward Metastasis-On-A-Chip: The Ultimate Model?

Author

Ruiz-Espigares, Jesús, Nieto, Daniel, Moroni, Lorenzo, Jiménez, Gema, Marchal, Juan Antonio, [Ruiz-Espigares,J, Jiménez,G, Marchal,JA] Biopathology and Regenerative Medicine Institute (IBIMER) Centre for Biomedical Research (CIBM) University of Granada, Granada, Spain. [Ruiz-Espigares,J, Marchal,JA] Department of Human Anatomy and Embryology Faculty of Medicine University of Granada, Granada, Spain. [Ruiz-Espigares,J, Marchal,JA] Excellence Research Unit 'Modeling Nature' (MNat) University of Granada, Granada, Spain. [Ruiz-Espigares,J, Marchal,JA] Biosanitary Research Institute of Granada (ibs.GRANADA) University Hospitals of Granada-University of Granada, Granada, Spain. [Nieto,D] Photonics4life Research Group Applied Physics Department Faculty of Physics University of Santiago de Compostela, Santiago de Compostela, Spain. [Nieto,D, Moroni,L] Complex Tissue Regeneration Department MERLN Institute for Technology Inspired Regenerative Medicine Universiteitssingel 40, Maastricht, The Netherlands., and This research was funded by the Consejería de Economía, Conocimiento, Empresas y Universidad, Junta de Andalucía (projects no. P18-FR-2470, B-CTS-230-UGR18, and OMM17/6109/UGR, FEDER Funds), the Instituto de Salud Carlos III (projects no. PIE16-00045 and DTS19/00145, FEDER Funds), the Consejería de Salud y Familias de la Junta de Andalucía (project no. PIN-0224-2019), and from the Chair 'Doctors Galera-Requena in cancer stem cell research' (CMC-CTS963).

Subjects
3D bioprinting, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Equipment and Supplies::Electrical Equipment and Supplies::Micro-Electrical-Mechanical Systems::Lab-On-A-Chip Devices [Medical Subject Headings], Analytical, Diagnostic and Therapeutic Techniques and Equipment::Diagnosis::Early Diagnosis::Early Detection of Cancer [Medical Subject Headings], Phenomena and Processes::Cell Physiological Phenomena::Cellular Microenvironment::Tumor Microenvironment [Medical Subject Headings], Diseases::Neoplasms [Medical Subject Headings], Metastasis-on-a-chip, Bioimpresión, Tumor-on-a-chip, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Bioprinting [Medical Subject Headings], Metastasis, Impresión tridimensional, Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings], Microambiente tumoral, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Rheology::Microfluidics [Medical Subject Headings], Analytical, Diagnostic and Therapeutic Techniques and Equipment::Therapeutics::Clinical Protocols [Medical Subject Headings], Information Science::Information Science::Communications Media::Publications::Review Literature as Topic [Medical Subject Headings], Cancer
Abstract

For decades, several attempts have been made to obtain a mimetic model for the study of metastasis, the reason of most of deaths caused by cancer, in order to solve the unknown phenomena surrounding this disease. To better understand this cellular dissemination process, more realistic models are needed that are capable of faithfully recreating the entire and essential tumor microenvironment (TME). Thus, new tools known as tumor-on-a-chip and metastasis-on-a-chip have been recently proposed. These tools incorporate microfluidic systems and small culture chambers where TME can be faithfully modeled thanks to 3D bioprinting. In this work, a literature review has been developed about the different phases of metastasis, the remaining unknowns and the use of new models to study this disease. The aim is to provide a global vision of the current panorama and the great potential that these systems have for in vitro translational research on the molecular basis of the pathology. In addition, these models will allow progress toward a personalized medicine, generating chips from patient samples that mimic the original tumor and the metastatic process to perform a precise pharmacological screening by establishing the most appropriate treatment protocol. Yes

Published
2021

12. Antioxidants for the Treatment of Breast Cancer: Are We There Yet?

Author

Griñan-Lison, Carmen, Blaya-Cánovas, Jose L., López-Tejada, Araceli, Ávalos-Moreno, Marta, Navarro-Ocón, Alba, Cara, Francisca E., González-González, Adrián, Lorente, Jose A., Marchal, Juan A., Granados-Principal, Sergio, [Griñan-Lison,C, Marchal,JA] Centre for Biomedical Research (CIBM), Biopathology and Regenerative Medicine Institute (IBIMER), University of Granada, Granada, Spain. [Griñan-Lison,C, Marchal,JA, Granados-Principal,S] Instituto de Investigación Biosanitaria Ibs.GRANADA, University Hospitals of Granada-University of Granada, Granada, Spain. [Griñan-Lison,C, Marchal,JA] Excellence Research Unit 'Modeling Nature' (MNat), University of Granada, Granada, Spain. [Griñan-Lison,C, Blaya-Cánovas,JL, López-Tejada,A, Ávalos-Moreno,M, Navarro-Ocón,A, Cara,FE, González-González,A, Lorente,JA, Granados-Principal,S] GENYO, Centre for Genomics and Oncological Research, Pfizer/University of Granada/Andalusian Regional Government, Granada, Spain. [Lorente,JA] Department of Legal Medicine, School of Medicine, University of Granada, Granada, Spain. [Marchal,JA] Department of Human Anatomy and Embryology, School of Medicine, University of Granada, Granada, Spain. [Granados-Principal,S] Department of Biochemistry and Molecular Biology II, School of Pharmacy, University of Granada, Granada, Spain., and This work and S.G.-P. are funded by Instituto de Salud Carlos III (CP19/00029, CP14/00197, PI19/01533, PI15/00336) and PIE16-00045 (J.A.M.), Ministerio de Ciencia, Innovación y niversi dades (RTI2018-101309-B-C22) (J.A.M.), and European Regional Development Fund (FEDER Funds,European Union) (S.G.-P., J.A.M.). C.G.-L. is funded by Junta de Andalucía, Consejería de Salud y Familias (RH-0139-2020), J.L.B.-C. is supported by Fundación Científica Asociación Española Contra el Cáncer, Junta Provincial de Jaén (AECC) (PRDJA19001BLAY), and A.L.-T. is funded by Ministerio de Ciencia, Innovación y Universidades (FPU19/04450).

Subjects
Células madre neoplásicas, Especies reactivas de oxígeno, Cancer stem cells, Ensayos clínicos como asunto, Phenomena and Processes::Metabolic Phenomena::Metabolism::Energy Metabolism::Oxidation-Reduction [Medical Subject Headings], Antioxidantes, Diseases::Neoplasms::Neoplastic Processes::Carcinogenesis [Medical Subject Headings], Diseases::Skin and Connective Tissue Diseases::Skin Diseases::Breast Diseases::Breast Neoplasms [Medical Subject Headings], Adjuvant therapy, Antioxidants, Cancer prevention, Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings], Breast cancer, Clinical trials, Chemicals and Drugs::Chemical Actions and Uses::Specialty Uses of Chemicals::Protective Agents::Antioxidants [Medical Subject Headings], Check Tags::Female [Medical Subject Headings], Chemicals and Drugs::Inorganic Chemicals::Oxygen Compounds::Reactive Oxygen Species [Medical Subject Headings], Neoplasias de la mama, Reactive oxygen species, Phenomena and Processes::Metabolic Phenomena::Metabolism::Oxidative Stress [Medical Subject Headings], Phenomena and Processes::Physiological Phenomena::Physiological Processes::Homeostasis [Medical Subject Headings]
Abstract

Breast cancer is the most frequent cancer and the leading cause of cancer death in women. Oxidative stress and the generation of reactive oxygen species (ROS) have been related to cancer progression. Compared to their normal counterparts, tumor cells show higher ROS levels and tight regulation of REDOX homeostasis to maintain a low degree of oxidative stress. Traditionally antioxidants have been extensively investigated to counteract breast carcinogenesis and tumor progression as chemopreventive agents; however, there is growing evidence indicating their potential as adjuvants for the treatment of breast cancer. Aimed to elucidate whether antioxidants could be a reality in the management of breast cancer patients, this review focuses on the latest investigations regarding the ambivalent role of antioxidants in the development of breast cancer, with special attention to the results derived from clinical trials, as well as their potential use as plausible agents in combination therapy and their power to ameliorate the side effects attributed to standard therapeutics. Data retrieved herein suggest that antioxidants play an important role in breast cancer prevention and the improvement of therapeutic efficacy; nevertheless, appropriate patient stratification based on "redoxidomics" or tumor subtype is mandatory in order to define the dosage for future standardized and personalized treatments of patients. Yes

Published
2021

13. Metabolomic profile of cancer stem cell-derived exosomes from patients with malignant melanoma

Author

Palacios-Ferrer, José Luis, García-Ortega, María Belén, Gallardo-Gómez, María, García, María Ángel, Díaz, Caridad, Boulaiz, Houria, Valdivia, Javier, Jurado, José Miguel, Almazan-Fernandez, Francisco M, Arias-Santiago, Salvador, Amezcua, Víctor, Peinado, Héctor, Vicente, Francisca, Pérez Del Palacio, José, Marchal, Juan A, Peinado Selgas, Hector, [Palacios-Ferrer,JL, García-Ortega,MB, García,MÁ, Boulaiz,H, Marchal,JA] Biopathology and Regenerative Medicine Institute (IBIMER), Centre for Biomedical Research (CIBM), University of Granada, Spain. [Palacios-Ferrer,JL, Marchal,JA] Department of Human Anatomy and Embryology, Faculty of Medicine, University of Granada, Spain. [Palacios-Ferrer,JL, Valdivia,J, Jurado,JM, Almazan-Fernandez,FM, Arias-Santiago,S, Amezcua,V, Marchal,JA] Instituto de Investigacion Biosanitaria de Granada (ibs.GRANADA), Spain. [Palacios-Ferrer,JL, Marchal,JA] Excellence Research Unit ‘Modeling Nature’ (MNat), University of Granada, Spain. [García-Ortega,MB, Amezcua,V] Department of Oncology, Virgen de las Nieves University Hospital, Granada, Spain. [Gallardo-Gómez,M] Department of Biochemistry, Genetics and Immunology, Singular Research Centre of Galicia (CINBIO), University of Vigo, Spain. [García,MÁ] Department of Biochemistry 3 and Immunology, Faculty of Medicine, University of Granada, Spain. [Díaz,C, Vicente,F, Pérez Del Palacio,J] Fundacion MEDINA, Centro de Excelencia en Investigación de Medicamentos Innovadores en Andalucía, Parque Tecnológico Ciencias de la Salud, Granada, Spain. [Jurado,JM] Department of Oncology, San Cecilio University Hospital, Granada, Spain. [Almazan-Fernandez,FM] Department of Dermatology, San Cecilio University Hospital, Granada, Spain. [Arias-Santiago,S] Department of Dermatology, Virgen de las Nieves University Hospital, Granada, Spain. [Arias-Santiago,S] Department of Medicine, Faculty of Medicine, University of Granada, Spain. [Peinado,H] Microenvironment and Metastasis Laboratory, Molecular Oncology Program, Spanish National Cancer Research Centre (CNIO), Madrid, Spain., This research was supported by the Ministerio de Ciencia, Innovacion y Universidades (MICIU, projectnos. MAT2015-62644.C2.2.R and RTI2018-101309-B C2, FEDER Funds), by the Instituto de Salud Carlos III (PIE16-00045), by Consejería de Economía, Conocimiento, Empresas y Universidad de la Junta de Andalucía and European Regional Development Fund (ERDF), ref. SOMM17/6109/UGR (UCE-PP2017-3), and by the Chair ‘Doctors Galera-Requena in cancer stem cell research’ (CMC-CTS963). MEDINA authors disclosed the receipt of financial support from Fun dacion MEDINA, a public-private partnership of Merck Sharp and Dohme de Espana S.A./Universidad ~ de Granada/Junta de Andalucía., Instituto de Salud Carlos III, Ministerio de Ciencia, Innovación y Universidades (España), Unión Europea. Comisión Europea, Regional Government of Andalusia (España), Instituto de Salud Carlos III - ISCIII, Ministerio de Ciencia, Innovacion y Universidades (MICIU), European Commission, and Junta de Andalucía (España)

Subjects
cancer stem cells, 0301 basic medicine, Anatomy::Cells::Cells, Cultured::Cell Line::Cell Line, Tumor [Medical Subject Headings], Cancer Research, Skin Neoplasms, Anatomy::Cells::Stem Cells::Neoplastic Stem Cells [Medical Subject Headings], Disease, HEPATOCELULAR CARCINOMA, Exosomes, DISEASE, Metastasis, Exosomas, Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings], 0302 clinical medicine, TUMOR, 3207.03 Carcinogénesis, Melanoma, Research Articles, MICROVESICLES, PLASMA, Malignant melanoma, Cancer stem cells, Disciplines and Occupations::Natural Science Disciplines::Biological Science Disciplines::Biochemistry::Metabolomics [Medical Subject Headings], General Medicine, Metabolómica, lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens, metabolomics, Oncology, 030220 oncology & carcinogenesis, Neoplastic Stem Cells, Molecular Medicine, Stem cell, Anatomy::Cells::Cellular Structures::Intracellular Space::Cytoplasm::Cytoplasmic Structures::Organelles::Cytoplasmic Vesicles::Transport Vesicles::Exosomes [Medical Subject Headings], Research Article, malignant melanoma, exosomes, lcsh:RC254-282, 03 medical and health sciences, Metabolomics, 2302 Bioquímica, Cancer stem cell, Cell Line, Tumor, REVEALS, Genetics, medicine, Humans, Diseases::Neoplasms::Neoplasms by Histologic Type::Nevi and Melanomas::Melanoma [Medical Subject Headings], Células madre neoplásicas, THERAPEUTIC TARGET, business.industry, Diseases::Neoplasms::Neoplasms by Site::Skin Neoplasms [Medical Subject Headings], biomarkers, medicine.disease, Microvesicles, 030104 developmental biology, 2405 Biometría, Biomarcadores, Cancer research, Skin cancer, business, Biomarkers
Abstract

Malignant melanoma (MM) is the most aggressive and life-threatening form of skin cancer. It is characterized by an extraordinary metastasis capacity and chemotherapy resistance, mainly due to melanoma cancer stem cells (CSCs). To date, there are no suitable clinical diagnostic, prognostic or predictive biomarkers for this neoplasia. Therefore, there is an urgent need for new MM biomarkers that enable early diagnosis and effective disease monitoring. Exosomes represent a novel source of biomarkers since they can be easily isolated from different body fluids. In this work, a primary patient-derived MM cell line enriched in CSCs was characterized by assessing the expression of specific markers and their stem-like properties. Exosomes derived from CSCs and serums from patients with MM were characterized, and their metabolomic profile was analysed by highresolution mass spectrometry (HRMS) following an untargeted approach and applying univariate and multivariate statistical analyses. The aim of this study was to search potential biomarkers for the diagnosis of this disease. Our results showed significant metabolomic differences in exosomes derived from MM CSCs compared with those from differentiated tumour cells and also in serum-derived exosomes from patients with MM compared to those from healthy controls. Interestingly, we identified similarities between structural lipids differentially expressed in CSC-derived exosomes and those derived from patients with MM such as the glycerophosphocholine PC 16:0/0:0. To our knowledge, this is the first metabolomic-based study aimed at characterizing exosomes derived from melanoma CSCs and patients’ serum in order to identify potential biomarkers for MM diagnosis. We conclude that metabolomic characterization of CSC-derived exosomes sets an open door to the discovery of clinically useful biomarkers in this neoplasia., MICIU FPU15/03682 FPU15/02350, Ministerio de Ciencia, Innovación y Universidades (MICIU) MAT2015-62644.C2.2.R RTI2018-101309-BC2, Instituto de Salud Carlos III PIE16-00045, Junta de Andalucía SOMM17/6109/UGR (UCE-PP2017-3), European Union (EU) SOMM17/6109/UGR (UCE-PP2017-3), Chair 'Doctors Galera-Requena in cancer stem cell research' CMC-CTS963, Fundación MEDINA

Published
2020

14. The Inhibitory Role of miR-486-5p on CSC Phenotype Has Diagnostic and Prognostic Potential in Colorectal Cancer

Author

Federico Attene, Sabine Atzmüller, Barbara Pardini, Fabrizio Scognamillo, Andrea Pisano, Grazia Fenu, Jesùs Peña-Martin, Maria Giuliana Solinas, Cristiano Farace, Juan A. Marchal, David Bernhard, Gabriele Ibba, Gema Jiménez, Johannes Pröll, Giovanni Fiorito, Roberto Madeddu, Carmen Griñán-Lisón, Alessio Naccarati, [Pisano,A, Farace,C, Fiorito,G, Fenu,G, Ibba,G, Solinas,MG, Madeddu,R] Department of Biomedical Sciences, University of Sassari, Sassari, Italy. [Pisano,A, Madeddu,R] National Institute of Biostructures and Biosystems, Rome, Italy. [Pisano,A, Griñan-Lison,C, Jiménez,G, Peña-Martin,J, Marchal,JA] Centre for Biomedical Research (CIBM), Biopathology and Regenerative Medicine Institute (IBIMER), University of Granada, Granada, Spain. [Griñan-Lison,C, Marchal,JA] Instituto de Investigación Biosanitaria Ibs.GRANADA, Organization University Hospitals of Granada, Granada, Spain. [Griñan-Lison,C, Jiménez,C, Marchal,JA] Excellence Research Unit Modeling Nature (MNat), University of Granada, Granada, Spain. [Fiorito,G] MRC Centre for Environment and Health, Imperial College London, Norfolk Place, London, UK. [Griñan-Lison,C, Marchal,JA] Department of Human Anatomy and Embryology, Faculty of Medicine, University of Granada, Granada, Spain. [Scognamillo,F, Attene,F] O.U. of Surgery I (Surgical Pathology), A.O.U. Sassari, Sassari, Italy. [Naccarati,A, Pardini,B] Molecular Epidemiology and Exposome Research Unit, Italian Institute for Genomic Medicine (IIGM), c/o IRCCS Candiolo, Candiolo, Torino, Italy. [Naccarati,A, Pardini,B] Molecular Epidemiology and Exposome Research Unit Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy. [Pröll,J] Austrian Cluster for Tissue Regeneration, Vienna, Austria. [Pröll,J, Atzmüller,S] Center for Medical Research, Johannes Kepler University, Linz, Austria. [Pröll,J] Red Cross Blood Transfusion Service, Linz, Austria. [Bernhard,D] Division of Pathophysiology, Institute of Physiology and Pathophysiology, Medical Faculty, Johannes Kepler University, Linz, Austria., This research was funded by: (1) Instituto de Salud Carlos III (PIE16-00045 and DTS19/00145), by Consejería de Economía, Conocimiento, Empresas y Universidad de la Junta de Andalucía and European Regional Development Fund (ERDF), ref. SOMM17/6109/UGR (UCE-PP2017-3), and by the Chair 'Doctors Galera-Requena in cancer stem cell research' (CMC-CTS963), and (2) Fondazione Banco di Sardegna (FBS), Cagliari, Italy.

Subjects
0301 basic medicine, cancer stem cells, Cancer Research, miR-486-5p, Colorectal cancer, Cell, Anatomy::Cells::Stem Cells::Neoplastic Stem Cells [Medical Subject Headings], Metastasis, Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings], 0302 clinical medicine, Anatomy::Cells::Cells, Cultured::Cell Line [Medical Subject Headings], Neoplasias colorrectales, Serum and stool biomarkers, education.field_of_study, MicroARNs, Cancer stem cells, serum and stool biomarkers, lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens, Persons::Persons::Patients [Medical Subject Headings], medicine.anatomical_structure, Oncology, Metástasis de la neoplasia, 030220 oncology & carcinogenesis, MiR-486-5p, Population, colorectal cancer, lcsh:RC254-282, Article, 03 medical and health sciences, Cancer stem cell, microRNA, Diseases::Neoplasms::Neoplastic Processes::Neoplasm Metastasis [Medical Subject Headings], medicine, Chemicals and Drugs::Biological Factors::Biological Markers [Medical Subject Headings], metastasis, Epigenetics, education, Células madre neoplásicas, business.industry, Cancer, Diseases::Neoplasms::Neoplasms by Site::Digestive System Neoplasms::Gastrointestinal Neoplasms::Intestinal Neoplasms::Colorectal Neoplasms [Medical Subject Headings], Chemicals and Drugs::Nucleic Acids, Nucleotides, and Nucleosides::Nucleic Acids::RNA::RNA, Antisense::MicroRNAs [Medical Subject Headings], medicine.disease, 030104 developmental biology, Biomarcadores, Cancer research, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Diagnosis::Prognosis [Medical Subject Headings], Phenomena and Processes::Genetic Phenomena::Genetic Structures::Genome::Genome Components::Genes::Genes, Neoplasm::Oncogenes [Medical Subject Headings], business
Abstract

Colorectal cancer (CRC) is the third most frequent cancer worldwide and the second cause of cancer deaths. Increasing evidences supports the idea that the poor prognosis of patients is related to the presence of cancer stem cells (CSCs), a cell population able to drive cancer recurrence and metastasis. The deregulation of microRNAs (miRNAs) plays a role in the formation of CSC. We investigated the role of hsa-miR-486-5p (miR-486-5p) in CRC, CSCs, and metastasis, in order to reach a better understanding of the biomolecular and epigenetic mechanisms mir-486-5p-related. The expression of miR-486-5p was investigated in three di erent matrices from CRC patients and controls and in CSCs obtained from the CRC cell lines HCT-116, HT-29, and T-84. In the human study, miR-486-5p was up-regulated in serum and stool of CRC patients in comparison with healthy controls but down-regulated in tumor tissue when compared with normal mucosa. miR-486-5p was also down-regulated in the sera of metastatic patients. In vitro, miR-486-5p was down-regulated in CSC models and it induced an inhibitory e ect on stem factors and oncogenes in the main pathways of CSCs. Our results provide a step forward in understanding the role of mir-486-5p in CRC and CSC, and suggest that further studies are needed to investigate its diagnostic and prognostic power, possibly in combination with other biomarkers., Instituto de Salud Carlos III PIE16-00045 DTS19/00145, Junta de Andalucía, European Union (EU) SOMM17/6109/UGR (UCE-PP2017-3), Chair "Doctors Galera-Requena in cancer stem cell research" CMC-CTS963, Fondazione Banco di Sardegna

Published
2020
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15. Evaluation of Glycerylphytate Crosslinked Semi- and Interpenetrated Polymer Membranes of Hyaluronic Acid and Chitosan for Tissue Engineering

Author

María Rosa Aguilar, Juan A. Marchal, Julio San Román, María Luisa López-Donaire, José Luis Pedraz, Patricia Gálvez-Martín, Blanca Vázquez-Lasa, Gema Jiménez, Elena López-Ruiz, Ana Mora-Boza, [Mora-Boza,A, López-Donaire,ML, Aguilar,MR, Vázquez-Lasa,B, San Román,J] Institute of Polymer Science and Technology, ICTP-CSIC, Madrid, Spain. [Mora-Boza,A, Pedraz,JL, San Román,J] CIBER-BBN, Health Institute Carlos III, Madrid, Spain. [López-Ruiz,E, Jiménez,G, Marchal,JA] Biopathology and Regenerative Medicine Institute (IBIMER), Centre for Biomedical Research, University of Granada, Granada, Spain. [López-Ruiz,E, Marchal,JA] Instituto de Investigación Biosanitaria de Granada (ibs.GRANADA), University Hospitals of Granada University of Granada, Granada, Spain. [López-Ruiz,E, Jiménez,G] Department of Health Sciences, University of Jaén, Jaén, Spain. [López-Ruiz,E, Marchal,JA] Excellence Research Unit 'Modeling Nature' (MNat), University of Granada, Granada, Spain. [Marchal,JA] Department of Human Anatomy and Embryology, Faculty of Medicine, University of Granada, Granada, Spain. [Pedraz,JL] NanoBioCel Group, Laboratory of Pharmaceutics, University of the Basque Country (UPV/EHU), School of Pharmacy, Vitoria-Gasteiz, Spain. [Gálvez-Martín,P] R&D Human Health, Bioibérica S.A.U., Spain., and The authors thanks to 'La Caixa' Foundation (ID 100010434), which supported Ana Mora-Boza (scholarship code LCF/BQ/ES16/11570018) and to the Spanish Ministry of Economy and Competitiveness for financial support (project RTC-2016-5451-1) and the Fundación Mutua Madrileña (project FMM-AP17196-2019). M. R. Aguilar and B. Vázquez-Lasa are members of the SusPlast platform (Interdisciplinary Platform for Sustainable Plastics towards a Circular Economy) from the Spanish National Research Council (CSIC).

Subjects
Polímeros, Polymers and Plastics, Polymers, Technology and Food and Beverages::Technology, Industry, and Agriculture::Manufactured Materials::Biomedical and Dental Materials::Polymers::Plastics::Resins, Synthetic::Acrylic Resins::Polymethacrylic Acids::Methylmethacrylates [Medical Subject Headings], Synthetic membrane, Ionic bonding, 02 engineering and technology, 01 natural sciences, Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings], Chitosan, chemistry.chemical_compound, Tissue engineering, Hyaluronic acid, mesenchymal stem cell, Disciplines and Occupations::Natural Science Disciplines::Biological Science Disciplines::Biotechnology::Biomimetics [Medical Subject Headings], glycerylphytate, 021001 nanoscience & nanotechnology, Quitosano, Membrane, Semi-IPN, semi-IPN, methacrylated hyaluronic acid, 0210 nano-technology, Interpenetrated polymer network, Biocompatibility, Células madre mesenquimatosas, interpenetrated polymer network, Chemicals and Drugs::Carbohydrates::Polysaccharides::Glycosaminoglycans::Hyaluronic Acid [Medical Subject Headings], macromolecular substances, 010402 general chemistry, Methacrylate, Article, lcsh:QD241-441, lcsh:Organic chemistry, Glycerylphytate, Methacrylated hyaluronic acid, Human Mesenchymal Stem Cells, Chemicals and Drugs::Complex Mixtures::Colloids::Gels::Hydrogels [Medical Subject Headings], Phenomena and Processes::Cell Physiological Phenomena::Cell Physiological Processes::Cell Growth Processes::Cell Proliferation [Medical Subject Headings], Technology and Food and Beverages::Technology, Industry, and Agriculture::Engineering::Bioengineering::Cell Engineering::Tissue Engineering [Medical Subject Headings], Chemicals and Drugs::Macromolecular Substances::Polymers::Biopolymers::Chitin::Chitosan [Medical Subject Headings], mesenchymal stem cel, technology, industry, and agriculture, General Chemistry, Chemicals and Drugs::Macromolecular Substances::Polymers [Medical Subject Headings], 0104 chemical sciences, chemistry, Chemical engineering, Ingeniería de tejidos
Abstract

In the present study, semi- and interpenetrated polymer network (IPN) systems based on hyaluronic acid (HA) and chitosan using ionic crosslinking of chitosan with a bioactive crosslinker, glycerylphytate (G1Phy), and UV irradiation of methacrylate were developed, characterized and evaluated as potential supports for tissue engineering. Semi- and IPN systems showed significant di erences between them regarding composition, morphology, and mechanical properties after physicochemical characterization. Dual crosslinking process of IPN systems enhanced HA retention and mechanical properties, providing also flatter and denser surfaces in comparison to semi-IPN membranes. The biological performance was evaluated on primary human mesenchymal stem cells (hMSCs) and the systems revealed no cytotoxic e ect. The excellent biocompatibility of the systems was demonstrated by large spreading areas of hMSCs on hydrogel membrane surfaces. Cell proliferation increased over time for all the systems, being significantly enhanced in the semi-IPN, which suggested that these polymeric membranes could be proposed as an e ective promoter system of tissue repair. In this sense, the developed crosslinked biomimetic and biodegradable membranes can provide a stable and amenable environment for hMSCs support and growth with potential applications in the biomedical field., La Caixa Foundation 100010434 LCF/BQ/ES16/11570018, Spanish Ministry of Economy and Competitiveness RTC-2016-5451-1, Instituto de Salud Carlos III FMM-AP17196-2019

Published
2020

16. Cancer stem cell secretome in the tumor microenvironment: a key point for an effective personalized cancer treatment

Author

López de Andrés, Julia, Griñán-Lisón, Carmen, Jiménez, Gema, Marchal, Juan Antonio, [López de Andrés,J, Griñán-Lisón,C, Jiménez,G, Marchal,JA] Biopathology and Regenerative Medicine Institute (IBIMER), Centre for Biomedical Research (CIBM), University of Granada,Granada, Spain. [López de Andrés,J, Marchal,JA] Instituto de Investigación Biosanitaria Ibs.GRANADA, University Hospitals of Granada-University of Granada, Granada, Spain. [López de Andrés,J, Marchal,JA] Excellence Research Unit 'Modeling Nature' (MNat), University of Granada, Granada, Spain. [Jiménez,G] Department of Health Sciences, University of Jaén, Jaén, Spain. [Marchal,JA] Department of Human Anatomy and Embryology, Faculty of Medicine, University of Granada, Granada, Spain., and This work was supported by grants from Consejería de Salud y Familias de la Junta de Andalucía (Project Numbers PIN-0224-2019), by the Ministerio de Ciencia, Innovación y Universidades Grant Number RTI2018-101309-B-C22 (FEDER Funds), by the Consejería de Economía, Conocimiento, Empresas y Universidad de la Junta de Andalucía (P18-FR-2470 and SOMM17/6109/UGR, FEDER Funds), by the Ministry of Economy and Competitiveness, Instituto de Salud Carlos III (FEDER funds, Projects Nos. PIE16/00045, DTS19/00143 and DTS17/00087), and from the Chair 'Doctors Galera-Requena in cancer stem cell research' (CMC-CTS963)

Subjects
Células madre neoplásicas, Anatomy::Cells::Cellular Structures::Extracellular Space::Extracellular Matrix [Medical Subject Headings], MicroARNs, Chemicals and Drugs::Biological Factors::Intercellular Signaling Peptides and Proteins::Cytokines::Interleukins [Medical Subject Headings], Cancer stem cells, Péptidos y proteínas de señalización intercelular, Interleukins, Phenomena and Processes::Cell Physiological Phenomena::Cellular Microenvironment::Tumor Microenvironment [Medical Subject Headings], Anatomy::Cells::Stem Cells::Neoplastic Stem Cells [Medical Subject Headings], Chemicals and Drugs::Nucleic Acids, Nucleotides, and Nucleosides::Nucleic Acids::RNA::RNA, Antisense::MicroRNAs [Medical Subject Headings], Diseases::Neoplasms [Medical Subject Headings], Exosomes, Chemicals and Drugs::Biological Factors::Intercellular Signaling Peptides and Proteins [Medical Subject Headings], Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings], Tumor microenvironment, Factores de crecimiento endotelial vascular, Microambiente tumoral, Interleucinas, miRNAs, Organisms::Eukaryota::Animals [Medical Subject Headings], Anatomy::Cells::Cellular Structures::Intracellular Space::Cytoplasm::Cytoplasmic Structures::Organelles::Cytoplasmic Vesicles::Transport Vesicles::Exosomes [Medical Subject Headings], Growth factors, Diseases::Pathological Conditions, Signs and Symptoms::Pathologic Processes::Metaplasia::Neovascularization, Pathologic [Medical Subject Headings], Secretome
Abstract

Cancer stem cells (CSCs) represent a tumor subpopulation responsible for tumor metastasis and resistance to chemo- and radiotherapy, ultimately leading to tumor relapse. As a consequence, the detection and eradication of this cell subpopulation represent a current challenge in oncology medicine. CSC phenotype is dependent on the tumor microenvironment (TME), which involves stem and differentiated tumor cells, as well as different cell types, such as mesenchymal stem cells, endothelial cells, fibroblasts and cells of the immune system, in addition to the extracellular matrix (ECM), different in composition to the ECM in healthy tissues. CSCs regulate multiple cancer hallmarks through the interaction with cells and ECM in their environment by secreting extracellular vesicles including exosomes, and soluble factors such as interleukins, cytokines, growth factors and other metabolites to the TME. Through these factors, CSCs generate and activate their own tumor niche by recruiting stromal cells and modulate angiogenesis, metastasis, resistance to antitumor treatments and their own maintenance by the secretion of different factors such as IL-6, VEGF and TGF-ß. Due to the strong influence of the CSC secretome on disease development, the new antitumor therapies focus on targeting these communication networks to eradicate the tumor and prevent metastasis, tumor relapse and drug resistance. This review summarizes for the first time the main components of the CSC secretome and how they mediate different tumor processes. Lastly, the relevance of the CSC secretome in the development of more precise and personalized antitumor therapies is discussed. Yes

Published
2020

17. Exosome: A New Player in Translational Nanomedicine

Author

Aheget, Houssam, Tristán-Manzano, María, Mazini, Loubna, Cortijo-Gutierrez, Marina, Galindo-Moreno, Pablo, Herrera, Concha, Martin, Francisco, Marchal, Juan Antonio, Benabdellah, Karim, [Aheget,H, Tristán-Manzano,M, Cortijo-Gutierrez,M, Martin,F, Benabdellah,K] Genomic Medicine Department, GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada (Andalusian Regional Government), Granada, Spain. [Mazini,L] Medical Application Interface Center, Mohammed VI Polytechnic University, Ben-Guerir, Morocco. [Galindo-Moreno,P] Oral Surgery and Implant Dentistry Department, School of Dentistry, University of Granada, Granada, Spain. [Herrera,C] Maimonides Institute of Biomedical Research in Cordoba (IMIBIC), Cordoba, Spain. [Herrera,C] Department of Haematology, Reina Sofía University Hospital, Cordoba, Spain. [Marchal,JA] Biomedical Research Institute, ibs. Granada, Granada, Spain. [Marchal,JA] Biopathology and Regenerative Medicine Institute (IBIMER), Centre for Biomedical Research (CIBM), University of Granada, Granada, Spain. [Marchal,JA] Department of Human Anatomy and Embryology, Faculty of Medicine, University of Granada, Granada, Spain. [Marchal,JA] Excellence Research Unit Modeling Nature (MNat), University of Granada, Granada, Spain., and This research was funded by the Spanish ISCIII Health Research Fund and the European Regional Development Fund (FEDER) through research grants PI12/01097, PI15/02015, PI18/00337 (F.M.), PIE16-00045 (J.A.M.), DTS19/00145 (J.A.M.) and PI18/00330 (K.B.), as well as by the Spanish Ministry of Science, Innovation and Universities (MICIU) through FEDER research grant RTI2018-101309-B-C2 (J.A.M.). The CECEyU and CSyF Councils of the Junta de Andalucía FEDER/European Cohesion Fund (FSE) provided the following research grants: 2016000073391-TRA, 2016000073332-TRA, PI-57069 and PAIDI-Bio326 (F.M.) and PI-0014-2016 (K.B). K.B. was also on a Nicolas Monardes Regional Ministry of Health contract (0006/2018). H.A. is supported by Research Excellence PhD Fellowship (2UAE2020) from the National Center for Scientific and Technical Research (CNRST). M.T.-M. is funded by the Spanish Ministry of Science and Innovation (SMSI) through an FPU16/05467 fellowship. M.C.-G. is funded by SMSI through a GJ fellowship (PEJ-2018-001760-A).

Subjects
Phenomena and Processes::Biological Phenomena [Medical Subject Headings], Anatomy::Cells::Cellular Structures::Intracellular Space::Cytoplasm::Cytoplasmic Structures::Organelles::Cytoplasmic Vesicles::Transport Vesicles::Endosomes::Multivesicular Bodies [Medical Subject Headings], Edición génica, Chemicals and Drugs::Lipids [Medical Subject Headings], Gene editing, Diseases::Neoplasms [Medical Subject Headings], Exosomes, Chemicals and Drugs::Nucleic Acids, Nucleotides, and Nucleosides::Nucleic Acids [Medical Subject Headings], Receptores quiméricos de antígenos, Neoplasias, Liquid biopsies, Exosomas, Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings], Biopsia líquida, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Therapeutics::Biological Therapy::Immunomodulation::Immunotherapy [Medical Subject Headings], CARs, Immunotherapy, Inmunoterapia, Anatomy::Cells::Cellular Structures::Intracellular Space::Cytoplasm::Cytoplasmic Structures::Organelles::Cytoplasmic Vesicles::Transport Vesicles::Exosomes [Medical Subject Headings], Chemicals and Drugs::Biological Factors::Biological Markers::Biomarkers, Pharmacological [Medical Subject Headings], Anatomy::Cells::Cellular Structures::Cell Membrane [Medical Subject Headings], Cancer
Abstract

Exosomes are extracellular vesicles released by the vast majority of cell types both in vivo and ex vivo, upon the fusion of multivesicular bodies (MVBs) with the cellular plasma membrane. Two main functions have been attributed to exosomes: their capacity to transport proteins, lipids and nucleic acids between cells and organs, as well as their potential to act as natural intercellular communicators in normal biological processes and in pathologies. From a clinical perspective, the majority of applications use exosomes as biomarkers of disease. A new approach uses exosomes as biologically active carriers to provide a platform for the enhanced delivery of cargo in vivo. One of the major limitations in developing exosome-based therapies is the difficulty of producing sufficient amounts of safe and efficient exosomes. The identification of potential proteins involved in exosome biogenesis is expected to directly cause a deliberate increase in exosome production. In this review, we summarize the current state of knowledge regarding exosomes, with particular emphasis on their structural features, biosynthesis pathways, production techniques and potential clinical applications. Yes

Published
2020

18. Validation of the 1,4-butanediol thermoplastic polyurethane as a novel material for 3D bioprinting applications

Author

Elvira Montañez, Juan A. Marchal, Elena López-Ruiz, Macarena Perán, Esmeralda Carrillo, Gema Jiménez, Juan de Vicente, Carlos Chocarro-Wrona, Cristina Antich, Patricia Gálvez-Martín, Daniel Martínez-Moreno, [Chocarro-Wrona,C, Antich,C, Jiménez,G, Martínez-Moreno,D, Carrillo,E, Perán,M, López-Ruiz,E, Marchal,JA] Biosanitary Research Institute of Granada (ibs.GRANADA), University Hospitals of Granada-University of Granada, Granada, Spain. [Chocarro-Wrona,C, Marchal,JA] Biopathology and Regenerative Medicine Institute (IBIMER), Centre for Biomedical Research (CIBM), University of Granada, Granada, Spain. [Chocarro-Wrona,C, Marchal,JA] Department of Human Anatomy and Embryology, Faculty of Medicine, University of Granada, Granada, Spain. [Chocarro-Wrona,C, de Vicente,J, Marchal,JA] Excellence Research Unit 'Modeling Nature' (MNat), University of Granada, Granada, Spain. [de Vicente,J] Department of Applied Physics, Faculty of Sciences, University of Granada, Granada, Spain. [Montañez,E] Biomedical Research Institute of Málaga (IBIMA), Málaga. [Montañez,E] Department of Orthopedic Surgery and Traumatology, Virgen de la Victoria University Hospital, Málaga, Spain. [Gálvez-Martín,P] Department of Pharmacy and Pharmaceutical Technology, School of Pharmacy, University of Granada, Granada, Spain. [Gálvez-Martín,P] Advanced Therapies Area, Bioibérica S.A.U, Barcelona, Spain. [Perán,M, López-Ruiz,E] Department of Health Sciences, University of Jaén, Jaén, Spain, Consejería de Economía, Innovación, Empresas y Universidad de la Junta de Andalucía and European Regional Development Fund, Grant/Award Number: SOMM17/6109/UGR, Ministerio de Economía, Indsutria y Competitividad. Gobierno de España, Grant/Award Numbers: MAT 2016-78778-R, PCIN-2015-051, Fundación Mutua Madrileña, Grant/Award Number: FMM-AP17196-2019., [Chocarro-Wrona, Carlos] Univ Granada, Univ Hosp Granada, Biosanitary Res Inst Granada Ibs GRANADA, Granada, Spain, [Antich, Cristina] Univ Granada, Univ Hosp Granada, Biosanitary Res Inst Granada Ibs GRANADA, Granada, Spain, [Jimenez, Gema] Univ Granada, Univ Hosp Granada, Biosanitary Res Inst Granada Ibs GRANADA, Granada, Spain, [Martinez-Moreno, Daniel] Univ Granada, Univ Hosp Granada, Biosanitary Res Inst Granada Ibs GRANADA, Granada, Spain, [Carrillo, Esmeralda] Univ Granada, Univ Hosp Granada, Biosanitary Res Inst Granada Ibs GRANADA, Granada, Spain, [Peran, Macarena] Univ Granada, Univ Hosp Granada, Biosanitary Res Inst Granada Ibs GRANADA, Granada, Spain, [Lopez-Ruiz, Elena] Univ Granada, Univ Hosp Granada, Biosanitary Res Inst Granada Ibs GRANADA, Granada, Spain, [Marchal, Juan Antonio] Univ Granada, Univ Hosp Granada, Biosanitary Res Inst Granada Ibs GRANADA, Granada, Spain, [Chocarro-Wrona, Carlos] Univ Granada, Ctr Biomed Res CIBM, Biopathol & Regenerat Med Inst IBIMER, Granada, Spain, [Antich, Cristina] Univ Granada, Ctr Biomed Res CIBM, Biopathol & Regenerat Med Inst IBIMER, Granada, Spain, [Jimenez, Gema] Univ Granada, Ctr Biomed Res CIBM, Biopathol & Regenerat Med Inst IBIMER, Granada, Spain, [Martinez-Moreno, Daniel] Univ Granada, Ctr Biomed Res CIBM, Biopathol & Regenerat Med Inst IBIMER, Granada, Spain, [Carrillo, Esmeralda] Univ Granada, Ctr Biomed Res CIBM, Biopathol & Regenerat Med Inst IBIMER, Granada, Spain, [Peran, Macarena] Univ Granada, Ctr Biomed Res CIBM, Biopathol & Regenerat Med Inst IBIMER, Granada, Spain, [Lopez-Ruiz, Elena] Univ Granada, Ctr Biomed Res CIBM, Biopathol & Regenerat Med Inst IBIMER, Granada, Spain, [Marchal, Juan Antonio] Univ Granada, Ctr Biomed Res CIBM, Biopathol & Regenerat Med Inst IBIMER, Granada, Spain, [Chocarro-Wrona, Carlos] Univ Granada, Fac Med, Dept Human Anat & Embryol, Granada, Spain, [Antich, Cristina] Univ Granada, Fac Med, Dept Human Anat & Embryol, Granada, Spain, [Martinez-Moreno, Daniel] Univ Granada, Fac Med, Dept Human Anat & Embryol, Granada, Spain, [Carrillo, Esmeralda] Univ Granada, Fac Med, Dept Human Anat & Embryol, Granada, Spain, [Marchal, Juan Antonio] Univ Granada, Fac Med, Dept Human Anat & Embryol, Granada, Spain, [Chocarro-Wrona, Carlos] Univ Granada, Excellence Res Unit Modeling Nat MNat, Granada, Spain, [de Vicente, Juan] Univ Granada, Excellence Res Unit Modeling Nat MNat, Granada, Spain, [Antich, Cristina] Univ Granada, Excellence Res Unit Modeling Nat MNat, Granada, Spain, [Jimenez, Gema] Univ Granada, Excellence Res Unit Modeling Nat MNat, Granada, Spain, [Martinez-Moreno, Daniel] Univ Granada, Excellence Res Unit Modeling Nat MNat, Granada, Spain, [Carrillo, Esmeralda] Univ Granada, Excellence Res Unit Modeling Nat MNat, Granada, Spain, [Peran, Macarena] Univ Granada, Excellence Res Unit Modeling Nat MNat, Granada, Spain, [Lopez-Ruiz, Elena] Univ Granada, Excellence Res Unit Modeling Nat MNat, Granada, Spain, [Marchal, Juan Antonio] Univ Granada, Excellence Res Unit Modeling Nat MNat, Granada, Spain, [de Vicente, Juan] Univ Granada, Dept Appl Phys, Fac Sci, Granada, Spain, [Montanez, Elvira] Biomed Res Inst Malaga IBIMA, Malaga, Spain, [Montanez, Elvira] Virgen Victoria Univ Hosp, Dept Orthoped Surg & Traumatol, Malaga, Spain, [Galvez-Martin, Patricia] Univ Granada, Sch Pharm, Dept Pharm & Pharmaceut Technol, Granada, Spain, [Galvez-Martin, Patricia] Bioiber SAU, Adv Therapies Area, Barcelona, Spain, [Peran, Macarena] Univ Jaen, Dept Hlth Sci, Jaen, Spain, [Lopez-Ruiz, Elena] Univ Jaen, Dept Hlth Sci, Jaen, Spain, Consejeria de Economia, Innovacion, Empresas y Universidad de la Junta de Andalucia, European Regional Development Fund, Ministerio de Economia, Indsutria y Competitividad. Gobierno de Espana, and Fundacion Mutua Madrilena

Subjects
Research Report, Pharmaceutical Science, MSCs, Elastómeros, law.invention, Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings], Tissue engineering, law, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Culture Techniques::Cell Engineering::Tissue Engineering [Medical Subject Headings], Technology and Food and Beverages::Technology, Industry, and Agriculture::Manufactured Materials::Biomedical and Dental Materials::Biocompatible Materials [Medical Subject Headings], 1,4-butanediol thermoplastic polyurethane, Poliuretanos, Biomaterial, Hydrogels, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Bioprinting [Medical Subject Headings], medicine.anatomical_structure, Elastomers, Chemicals and Drugs::Macromolecular Substances::Polymers::Polyesters [Medical Subject Headings], tissue engineering, Anatomy::Musculoskeletal System::Cartilage::Hyaline Cartilage::Cartilage, Articular [Medical Subject Headings], TP155-156, Collagen, Mesenchymal stem-cells, Chondrogenesis, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Diagnosis::Diagnostic Techniques and Procedures::Diagnostic Imaging::Microscopy::Microscopy, Confocal [Medical Subject Headings], Chemicals and Drugs::Organic Chemicals::Alcohols::Glycols::Butylene Glycols [Medical Subject Headings], Biotechnology, Biofabrication, Design, Materials science, Biocompatibility, Condrogénesis, Mechanical-properties, Biomedical Engineering, Células madre mesenquimatosas, Elastomer, RM1-950, Fabrication, Thermoplastic polyurethane, 1,4‐butanediol thermoplastic polyurethane, Chemical engineering, Pcl scaffolds, Phenomena and Processes::Cell Physiological Phenomena::Cell Physiological Processes::Cell Survival [Medical Subject Headings], Phenomena and Processes::Musculoskeletal and Neural Physiological Phenomena::Musculoskeletal Physiological Phenomena::Musculoskeletal Physiological Processes::Musculoskeletal Development::Chondrogenesis [Medical Subject Headings], medicine, Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Scleroproteins::Extracellular Matrix Proteins [Medical Subject Headings], Cartilage tissue, Phenomena and Processes::Physical Phenomena::Mechanical Phenomena::Elasticity [Medical Subject Headings], elastomer, 3D bioprinting, Matrix, Cartilage, Phenomena and Processes::Physical Phenomena::Mechanical Phenomena::Mechanical Processes::Weight-Bearing [Medical Subject Headings], Tissue enginering, Research Reports, Bioimpresión, Autologous chondrocyte implantation, Technology and Food and Beverages::Technology, Industry, and Agriculture::Manufactured Materials::Elastomers::Polyurethanes [Medical Subject Headings], Phenomena and Processes::Physical Phenomena::Mechanical Phenomena::Friction [Medical Subject Headings], Therapeutics. Pharmacology, Ingeniería de tejidos, TP248.13-248.65, Biomedical engineering
Abstract

Tissue engineering (TE) seeks to fabricate implants that mimic the mechanical strength, structure, and composition of native tissues. Cartilage TE requires the development of functional personalized implants with cartilage-like mechanical properties capable of sustaining high load-bearing environments to integrate into the surrounding tissue of the cartilage defect. In this study, we evaluated the novel 1,4-butanediol thermoplastic polyurethane elastomer (b-TPUe) derivative filament as a 3D bioprinting material with application in cartilage TE. The mechanical behavior of b-TPUe in terms of friction and elasticity were examined and compared with human articular cartilage, PCL, and PLA. Moreover, infrapatellar fat pad-derived human mesenchymal stem cells (MSCs) were bioprinted together with scaffolds. in vitro cytotoxicity, proliferative potential, cell viability, and chondrogenic differentiation were analyzed by Alamar blue assay, SEM, confocal microscopy, and RT-qPCR. Moreover, in vivo biocompatibility and host integration were analyzed. b-TPUe demonstrated a much closer compression and shear behavior to native cartilage than PCL and PLA, as well as closer tribological properties to cartilage. Moreover, b-TPUe bioprinted scaffolds were able to maintain proper proliferative potential, cell viability, and supported MSCs chondrogenesis. Finally, in vivo studies revealed no toxic effects 21 days after scaffolds implantation, extracellular matrix deposition and integration within the surrounding tissue. This is the first study that validates the biocompatibility of b-TPUe for 3D bioprinting. Our findings indicate that this biomaterial can be exploited for the automated biofabrication of artificial tissues with tailorable mechanical properties including the great potential for cartilage TE applications., Consejería de Economía, Innovación, Empresas y Universidad de la Junta de Andalucía SOMM17/6109/UGR, European Union (EU) SOMM17/6109/UGR, Ministerio de Economía, Industria y Competitividad. Gobierno de España MAT 2016-78778-R PCIN-2015-051, Instituto de Salud Carlos III FMM-AP17196-2019

Published
2020

19. Uncovering Tumour Heterogeneity through PKR and nc886 Analysis in Metastatic Colon Cancer Patients Treated with 5-FU-Based Chemotherapy

Author

María Ángel García, Alberto Mesa, Elisa L. J. Moya, Igor Zwir, Luis Javier Martinez-Gonzalez, Gabriel Lopez-Ordoño, María Belén Ortega-García, Eduardo Redondo-Cerezo, Javier Ángel García, Juan A. Marchal, Macarena Perán, Javier Luis Lopez-Hidalgo, V. Conde, Gema Jiménez, Coral del Val, Beatriz Rueda, [Ortega-García,MB, Moya,ELJ, Jiménez,G, Del Val,C, Zwir,I, Marchal,JA, García,MÁ] Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, Spain. [Ortega-García,MB, García,JÁ, Conde,V] Department of Oncology, Virgen de las Nieves University Hospital, Granada, Spain. [Ortega-García,MB, García,MÁ] Biopathology and Regenerative Medicine Institute (IBIMER), Centre for Biomedical Research, (CIBM) University of Granada, Granada, Spain. [Ortega-García,MB, Jiménez,J, Peran,M, García,MÁ] Excellence Research Unit 'Modelling Nature' (MNat), University of Granada, Granada, Spain. [Mesa,A, Zwir,I] Andalusian Research Institute in Data Science and Computational Intelligence (DaSCI Institute), Granada, Spain. [Rueda,B, Lopez-Hidalgo,JL] Department of Pathology, San Cecilio University Hospital, Granada, Spain. [Lopez-Ordoño,G] Department of Gastroenterology, Torrecardenas Hospital, Almería, Spain. [Redondo-Cerezo,E] Department of Gastroenterology, Virgen de las Nieves University Hospital, Granada, Spain. [Peran,M] Department of Health Sciences, University of Jaén, Jaen, Spain. [Martínez-González,LJ] GENYO: Centre for Genomics and Oncological Research: Pfizer/University of Granada/Andalusian Regional Government, PTS, Granada, Spain. [Del Val,C, Zwir,I] Department of Computer Science and Artificial Intelligence, University of Granada, Granada, Spain. [Zwir,I] Department of Psychiatry, Washington University School of Medicine, St Louis, USA. [Marchal,JA] Department of Human Anatomy and Embryology, University of Granada, Granada, Spain. [García,MÁ] Department of Biochemistry and Molecular Biology III, University of Granada, Granada, Spain., This research was funded by the Instituto de Salud Carlos III (DTS15/00174, and PIE16-00045), by the Consejería de Economía, Conocimiento, Empresas y Universidad de la Junta de Andalucía and European Regional Development Fund (ERDF), references SOMM17/6109/UGR (UCE-PP2017-3) and (PI-0441-2014), and by the Chair 'Doctors Galera-Requena in cancer stem cell research' (CMC-CTS963). This research was also funded partially by RTI2018-098983-B-I00.

Subjects
0301 basic medicine, Oncology, Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Transferases::Phosphotransferases::Phosphotransferases (Alcohol Group Acceptor)::Protein Kinases [Medical Subject Headings], Cancer Research, ambispective study, Cluster of patients, Colorectal cancer, medicine.medical_treatment, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Therapeutics::Combined Modality Therapy::Chemotherapy, Adjuvant [Medical Subject Headings], Regulator, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Immunologic Techniques::Immunohistochemistry [Medical Subject Headings], Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings], 0302 clinical medicine, Neoplasias colorrectales, Clinical endpoint, Chemicals and Drugs::Heterocyclic Compounds::Heterocyclic Compounds, 1-Ring::Pyrimidines::Pyrimidinones::Uracil::Fluorouracil [Medical Subject Headings], non-coding nc886, lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens, Ambispective study, 030220 oncology & carcinogenesis, Immunohistochemistry, cluster of patients, medicine.medical_specialty, Phenomena and Processes::Mathematical Concepts::Algorithms [Medical Subject Headings], Non-coding nc886, Fluorouracilo, Tumour heterogeneity, Protein kinase PKR, colorectal cancer, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Epidemiologic Methods::Statistics as Topic::Cluster Analysis [Medical Subject Headings], lcsh:RC254-282, Article, 03 medical and health sciences, Internal medicine, medicine, Chemotherapy, 5-fluorouracil-based chemotherapy, business.industry, protein kinase PKR, Cancer, biomarkers, Diseases::Neoplasms::Neoplasms by Site::Digestive System Neoplasms::Gastrointestinal Neoplasms::Intestinal Neoplasms::Colorectal Neoplasms [Medical Subject Headings], medicine.disease, Protein kinase R, Biomarcadores, 030104 developmental biology, Quimioterapia, Proteína quinasa PKR, business, Biomarkers
Abstract

Colorectal cancer treatment has advanced over the past decade. The drug 5-fluorouracil is still used with a wide percentage of patients who do not respond. Therefore, a challenge is the identification of predictive biomarkers. The protein kinase R (PKR also called EIF2AK2) and its regulator, the non-coding pre-mir-nc886, have multiple e ects on cells in response to numerous types of stress, including chemotherapy. In this work, we performed an ambispective study with 197 metastatic colon cancer patients with unresectable metastases to determine the relative expression levels of both nc886 and PKR by qPCR, as well as the location of PKR by immunohistochemistry in tumour samples and healthy tissues (plasma and colon epithelium). As primary end point, the expression levels were related to the objective response to first-line chemotherapy following the response evaluation criteria in solid tumours (RECIST) and, as the second end point, with survival at 18 and 36 months. Hierarchical agglomerative clustering was performed to accommodate the heterogeneity and complexity of oncological patients’ data. High expression levels of nc886 were related to the response to treatment and allowed to identify clusters of patients. Although the PKR mRNA expression was not associated with chemotherapy response, the absence of PKR location in the nucleolus was correlated with first-line chemotherapy response. Moreover, a relationship between survival and the expression of both PKR and nc886 in healthy tissues was found. Therefore, this work evaluated the best way to analyse the potential biomarkers PKR and nc886 in order to establish clusters of patients depending on the cancer outcomes using algorithms for complex and heterogeneous data., This research was funded by the Instituto de Salud Carlos III (DTS15/00174; PIE16-00045), by the Consejería de Economía, Conocimiento, Empresas y Universidad de la Junta de Andalucía and European Regional Development Fund (ERDF), references SOMM17/6109/UGR (UCE-PP2017-3) and (PI-0441-2014), and by the Chair “Doctors Galera-Requena in cancer stem cell research” (CMC-CTS963). This research was also funded partially by RTI2018-098983-B-I00.

Published
2020
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20. Role of Mesenchymal Stromal Cells as Therapeutic Agents: Potential Mechanisms of Action and Implications in Their Clinical Use

Author

Jimenez-Puerta, Gonzalo José, Marchal, Juan Antonio, López-Ruiz, Elena, Gálvez-Martín, Patricia, [Jimenez-Puerta,GJ, Marchal,JA, López-Ruiz.E] Biosanitary Research Institute of Granada (ibs.GRANADA), University Hospitals of Granada-University of Granada, Granada, Spain. [Marchal,JA, López-Ruiz.E] Biopathology and Regenerative Medicine Institute (IBIMER), Centre for Biomedical Research (CIBM), University of Granada, Granada, Spain. [Marchal,JA] Department of Human Anatomy and Embryology, Faculty of Medicine, University of Granada, Granada, Spain. [Marchal,JA, López-Ruiz.E] Excellence Research Unit 'Modeling Nature' (MNat), University of Granada, Granada, Spain. [López-Ruiz.E] Department of Health Sciences, University of Jaén, Jaén, Spain. [Gálvez-Martín,P] Department of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of Granada, Granada, Spain. [Gálvez-Martín,P] R&D Human Health, Bioibérica S.A.U., Barcelona, Spain., and This work was supported by the Fundación Mutua Madrileña (project FMM-AP17196-2019) and by the Ministerio de Economía, Industria y Competitividad (FEDER funds, project RTC-2016-5451-1).

Subjects
Homing, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Therapeutics [Medical Subject Headings], Mesenchymal stromal cells, Anatomy::Hemic and Immune Systems::Immune System [Medical Subject Headings], Anatomy::Cells [Medical Subject Headings], Células madre mesenquimatosas, Mechanism of action, Immunomodulation, Tejidos, Células, Anatomy::Cells::Stem Cells::Multipotent Stem Cells::Mesenchymal Stromal Cells [Medical Subject Headings], Anatomy::Tissues [Medical Subject Headings], Movimiento celular, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Therapeutics::Biological Therapy::Immunomodulation [Medical Subject Headings], Analytical, Diagnostic and Therapeutic Techniques and Equipment::Therapeutics::Biological Therapy::Cell- and Tissue-Based Therapy [Medical Subject Headings]
Abstract

Due to the great therapeutic interest that involves the translation of mesenchymal stromal cells (MSCs) into clinical practice, they have been widely studied as innovative drugs, in order to treat multiple pathologies. MSC-based cell therapy involves the administration of MSCs either locally or systemically into the receptor body where they can traffic and migrate towards the affected tissue and participate in the process of healing. The therapeutic effects of MSCs compromise of different mechanisms such as the functional integration of differentiated MSCs into diseased host tissue after transplantation, their paracrine support, and their impact on the regulation of both the innate and the acquired immune system. Here, we establish and provide recent advances about the principal mechanisms of action through which MSCs can perform their activity and effect as a therapeutic tool. The purpose of this review is to examine and discuss the MSCs capacity of migration, their paracrine effect, as well as MSC-mediated modifications on immune cell responses. Yes

Published
2020

21. Validation of suitable normalizers for miR expression patterns analysis covering tumour heterogeneity

Author

Macarena Perán, Houria Boulaiz, Cynthia Morata-Tarifa, Juan A. Marchal, Carmen Griñán-Lisón, María Ángel García, Manuel Picon-Ruiz, [Morata-Tarifa,C, Picon-Ruiz,M, Griñan-Lison,C, Boulaiz,H, Perán,M, Garcia,MA, Marchal,JA] Biopathology and Medicine Regenerative Institute (IBIMER), University of Granada, Granada, Spain. [Morata-Tarifa,C, Marchal,JA] Biosanitary Institute of Granada (ibs.GRANADA), University Hospitals of Granada-University of Granada, Granada, Spain. [Morata-Tarifa,C, Picón-Ruiz,M] Braman Family Breast Cancer Institute, Sylvester Comprehensive Cancer Center, University of Miami, Miller School of Medicine, Miami, Florida, USA. [Boulaiz,H, Marchal,JA] Department of Human Anatomy and Embryology, University of Granada, Granada, Spain. [Perán,M] Department of Health Sciences, University of Jaén, Jaén, Spain. [Garcia,MA] Virgen de las Nieves University Hospital, Oncology Department, Oncology Unit, Granada, Spain., and This work was supported in part by grants from the Fundación Pública Andaluza Progreso y Salud, Consejería de Salud Junta de Andalucía in collaboration with JANNSSEN CILAG,S.A. (project numbers PI-0533-2014, PI-0441-2014), from the Ministerio de Economía y Competitividad (MINECO, FEDER funds, grant number MAT2015-62644-C2-2-R) from the Ministry of Economy and Competitiveness, Instituto de Salud Carlos III (FEDER funds, project DTS15/00174) and from the Chair 'Doctors Galera-Requena in cancer stem cell research'.

Subjects
0301 basic medicine, Chemicals and Drugs::Nucleic Acids, Nucleotides, and Nucleosides::Nucleic Acids::RNA::RNA, Nuclear::RNA, Small Nuclear [Medical Subject Headings], Colorectal cancer, medicine.disease_cause, Bioinformatics, 0302 clinical medicine, Anatomy::Cells::Cells, Cultured::Cell Line [Medical Subject Headings], Neoplasms, ARN nucleolar pequeño, Small nucleolar RNA, MicroARNs, Multidisciplinary, Línea celular, Diseases::Neoplasms::Neoplastic Processes::Carcinogenesis [Medical Subject Headings], Housekeeping gene, Gene Expression Regulation, Neoplastic, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Epidemiologic Methods::Epidemiologic Research Design::Reproducibility of Results [Medical Subject Headings], Disciplines and Occupations::Natural Science Disciplines::Biological Science Disciplines::Biology::Computational Biology [Medical Subject Headings], 030220 oncology & carcinogenesis, Neoplasias de la mama, Biología computacional, Carcinogénesis, Tumour heterogeneity, Chemicals and Drugs::Biological Factors::Biomarkers [Medical Subject Headings], Computational biology, Biology, Real-Time Polymerase Chain Reaction, Article, 03 medical and health sciences, Phenomena and Processes::Genetic Phenomena::Genetic Structures::Genome::Genome Components::Genes::Genes, Essential [Medical Subject Headings], Cell Line, Tumor, microRNA, Biomarkers, Tumor, medicine, Humans, Neoplasias del colon, Diseases::Neoplasms::Neoplasms by Site::Breast Neoplasms [Medical Subject Headings], Reacción en cadena en tiempo real de la polimerasa, Reproducibilidad de resultados, Chemicals and Drugs::Chemical Actions and Uses::Pharmacologic Actions::Therapeutic Uses::Antineoplastic Agents [Medical Subject Headings], Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Genetic Techniques::Nucleic Acid Amplification Techniques::Polymerase Chain Reaction::Real-Time Polymerase Chain Reaction [Medical Subject Headings], Computational Biology, Cancer, Oncogenes, medicine.disease, Antineoplásicos, Chemicals and Drugs::Nucleic Acids, Nucleotides, and Nucleosides::Antisense Elements (Genetics)::RNA, Antisense::MicroRNAs [Medical Subject Headings], MicroRNAs, 030104 developmental biology, Biomarcadores, Cancer cell, Diseases::Neoplasms::Neoplasms by Site::Digestive System Neoplasms::Gastrointestinal Neoplasms::Intestinal Neoplasms::Colorectal Neoplasms::Colonic Neoplasms [Medical Subject Headings], Phenomena and Processes::Genetic Phenomena::Genetic Structures::Genome::Genome Components::Genes::Genes, Neoplasm::Oncogenes [Medical Subject Headings], Carcinogenesis, Genes esenciales
Abstract

Oncogenic microRNAs (miRs) have emerged as diagnostic biomarkers and novel molecular targets for anti-cancer drug therapies. Real-time quantitative PCR (qPCR) is one of the most powerful techniques for analyzing miRs; however, the use of unsuitable normalizers might bias the results. Tumour heterogeneity makes even more difficult the selection of an adequate endogenous normalizer control. Here, we have evaluated five potential referenced small RNAs (U6, rRNA5s, SNORD44, SNORD24 and hsa-miR-24c-3p) using RedFinder algorisms to perform a stability expression analysis in i) normal colon cells, ii) colon and breast cancer cell lines and iii) cancer stem-like cell subpopulations. We identified SNORD44 as a suitable housekeeping gene for qPCR analysis comparing normal and cancer cells. However, this small nucleolar RNA was not a useful normalizer for cancer stem-like cell subpopulations versus subpopulations without stemness properties. In addition, we show for the first time that hsa-miR-24c-3p is the most stable normalizer for comparing these two subpopulations. Also, we have identified by bioinformatic and qPCR analysis, different miR expression patterns in colon cancer versus non tumour cells using the previously selected suitable normalizers. Our results emphasize the importance of select suitable normalizers to ensure the robustness and reliability of qPCR data for analyzing miR expression.

Published
2017

22. Low adherent cancer cell subpopulations are enriched in tumorigenic and metastatic epithelial-to-mesenchymal transition-induced cancer stem-like cells

Author

Margarita Aguilera, Carmen Griñán-Lisón, José Manuel Entrena, María Ángel García, Juan A. Marchal, Gema Jiménez, Cynthia Morata-Tarifa, Manuel Picon-Ruiz, [Morata-Tarifa,C, Jiménez,G, García,MA, Griñán-Lisón,C, Picon-Ruiz,M, Marchal,JA] Biopathology and Medicine Regenerative Institute (IBIMER), University of Granada, Granada, Spain. [Morata-Tarifa,C, Aguilera,M, Marchal,JA] Biosanitary Institute of Granada (ibs.GRANADA), University Hospitals of Granada-Univesity of Granada, Granada, Spain. [Jiménez,G, Marchal,JA] Department of Human Anatomy and Embryology, University of Granada, Granada, Spain. [García,MA] Department of Oncology, University Hospital Virgen de las Nieves, Granada, Spain. [Entrena,JM] Institute of Neuroscience, Biomedical Research Center, University of Granada, Granada, Spain. Animal Behavior Research Unit, Scientific Instrumentation Center, University of Granada, Granada, Spain. [Aguilera,M] Department of Microbiology, University of Granada, Granada, Spain. [Picon-Ruiz,M] Braman Family Breast Cancer Institute, Sylvester Comprehensive Cancer Center, University of Miami, Miller School of Medicine, Miami, Florida, USA., and This work was supported in part by grants from the Ministry of Economy and Competitiveness, Instituto de Salud Carlos III (FEDER funds, projects n°. PI10/02295 and PI10/02149), the Fundación Progreso y Salud, Consejería de Salud, Junta de Andalucía (Ministry of Health, Government of Andalucia, project number PI-0533-2014) and by Consejería Economía, Innovación, Ciencia y Empleo, Junta de Andalucia (Ministry of Economy, Innovation, Science and Employment, Government of Andalucia, excellence project number CTS-6568) providing a fellowship granted to GJ.

Subjects
0301 basic medicine, Pathology, Phenomena and Processes::Genetic Phenomena::Phenotype [Medical Subject Headings], Colorectal cancer, Claudinas, Ratones, Phenomena and Processes::Physiological Phenomena::Body Constitution::Body Weights and Measures::Body Mass Index [Medical Subject Headings], Anatomy::Cells::Stem Cells::Neoplastic Stem Cells [Medical Subject Headings], Índice de masa corporal, Metastasis, Anatomy::Body Regions::Breast [Medical Subject Headings], Mice, 0302 clinical medicine, Immunophenotyping, Vimentina, Neoplasms, Regulación hacia arriba, Organisms::Eukaryota::Animals [Medical Subject Headings], Caracoles, Cell Self Renewal, Neoplasm Metastasis, Side-Population Cells, Tumor Stem Cell Assay, Recurrencia local de neoplasia, Multidisciplinary, Transición epitelial-mesenquimal, Mama, Chemicals and Drugs::Heterocyclic Compounds::Heterocyclic Compounds, 2-Ring::Benzimidazoles [Medical Subject Headings], Cell Transformation, Neoplastic, Phenotype, 030220 oncology & carcinogenesis, Neoplasias de la mama, Neoplastic Stem Cells, Heterografts, Fenotipo, medicine.medical_specialty, Epithelial-Mesenchymal Transition, Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Hydrolases::Peptide Hydrolases::Endopeptidases::Serine Endopeptidases::Trypsin [Medical Subject Headings], Biology, Article, 03 medical and health sciences, Cancer stem cell, Cell Line, Tumor, Xenoinjertos, medicine, Cell Adhesion, Animals, Humans, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Equipment and Supplies::Transplants::Heterografts [Medical Subject Headings], Neoplasias del colon, Epithelial–mesenchymal transition, Diseases::Neoplasms::Neoplasms by Site::Breast Neoplasms [Medical Subject Headings], Células madre neoplásicas, Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Rodentia::Muridae::Murinae::Mice [Medical Subject Headings], Phenomena and Processes::Chemical Phenomena::Biochemical Phenomena::Biochemical Processes::Up-Regulation [Medical Subject Headings], Gene Expression Profiling, Cancer, Diseases::Neoplasms::Neoplastic Processes::Neoplasm Recurrence, Local [Medical Subject Headings], Organisms::Eukaryota::Animals::Invertebrates::Mollusca::Gastropoda::Snails [Medical Subject Headings], Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Glycoproteins::Membrane Glycoproteins::Cell Adhesion Molecules::Cadherins [Medical Subject Headings], medicine.disease, Tripsina, Chemicals and Drugs::Nucleic Acids, Nucleotides, and Nucleosides::Antisense Elements (Genetics)::RNA, Antisense::MicroRNAs [Medical Subject Headings], Disease Models, Animal, 030104 developmental biology, Tumor progression, Cancer cell, Animales, Diseases::Neoplasms::Neoplasms by Site::Digestive System Neoplasms::Gastrointestinal Neoplasms::Intestinal Neoplasms::Colorectal Neoplasms::Colonic Neoplasms [Medical Subject Headings], Cancer research, Phenomena and Processes::Cell Physiological Phenomena::Cell Physiological Processes::Cell Transdifferentiation::Epithelial-Mesenchymal Transition [Medical Subject Headings], Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Membrane Proteins::Claudins [Medical Subject Headings], Chemicals and Drugs::Macromolecular Substances::Polymers::Biopolymers::Intermediate Filament Proteins::Vimentin [Medical Subject Headings], Biomarkers, Cadherinas
Abstract

Cancer stem cells are responsible for tumor progression, metastasis, therapy resistance and cancer recurrence, doing their identification and isolation of special relevance. Here we show that low adherent breast and colon cancer cells subpopulations have stem-like properties. Our results demonstrate that trypsin-sensitive (TS) breast and colon cancer cells subpopulations show increased ALDH activity, higher ability to exclude Hoechst 33342, enlarged proportion of cells with a cancer stem-like cell phenotype and are enriched in sphere- and colony-forming cells in vitro. Further studies in MDA-MB-231 breast cancer cells reveal that TS subpopulation expresses higher levels of SLUG, SNAIL, VIMENTIN and N-CADHERIN while show a lack of expression of E-CADHERIN and CLAUDIN, being this profile characteristic of the epithelial-to-mesenchymal transition (EMT). The TS subpopulation shows CXCL10, BMI-1 and OCT4 upregulation, differing also in the expression of several miRNAs involved in EMT and/or cell self-renewal such as miR-34a-5p, miR-34c-5p, miR-21-5p, miR-93-5p and miR-100-5p. Furthermore, in vivo studies in immunocompromised mice demonstrate that MDA-MB-231 TS cells form more and bigger xenograft tumors with shorter latency and have higher metastatic potential. In conclusion, this work presents a new, non-aggressive, easy, inexpensive and reproducible methodology to isolate prospectively cancer stem-like cells for subsequent biological and preclinical studies., Ministry of Economy and Competitiveness, Instituto de Salud Carlos III (FEDER funds) PI10/02295 PI10/02149, Fundacion Progreso y Salud, Consejeria de Salud, Junta de Andalucia (Ministry of Health, Government of Andalucia) PI-0533-2014, Consejeria Economia, Innovacion, Ciencia y Empleo, Junta de Andalucia (Ministry of Economy, Innovation, Science and Employment, Government of Andalucia) CTS-6568

Published
2016

23. Knee Viscosupplementation: Cost-Effectiveness Analysis between Stabilized Hyaluronic Acid in a Single Injection versus Five Injections of Standard Hyaluronic Acid

Author

Francisco J. Estades-Rubio, Juan J. García-Vera, Juan A. Marchal, Victor Morales-Marcos, Alvaro Reyes-Martín, Macarena Perán, Elvira Montañez-Heredia, Mercedes García-Piriz, [Estades-Rubio, Francisco J.] Virgen Victoria Univ Hosp, Dept Orthoped Surg & Traumatol, E-29010 Malaga, Spain, [Garcia-Piriz, Mercedes] Virgen Victoria Univ Hosp, Dept Orthoped Surg & Traumatol, E-29010 Malaga, Spain, [Garcia-Vera, Juan J.] Virgen Victoria Univ Hosp, Dept Orthoped Surg & Traumatol, E-29010 Malaga, Spain, [Montanez-Heredia, Elvira] Virgen Victoria Univ Hosp, Dept Orthoped Surg & Traumatol, E-29010 Malaga, Spain, [Reyes-Martin, Alvaro] Hosp Reg Univ Carlos Haya Malaga, Dept Orthoped Surg & Traumatol, E-29010 Malaga, Spain, [Morales-Marcos, Victor] Hosp Quironsalud Malaga, Dept Orthoped Surg & Traumatol, E-29004 Malaga, Spain, [Peran, Macarena] Univ Jaen, Dept Hlth Sci, E-23071 Jaen, Spain, [Peran, Macarena] Univ Granada, Ctr Biomed Res, Biopathol & Regenerat Med Inst IBIMER, E-18071 Granada, Spain, [Marchal, Juan A.] Univ Granada, Ctr Biomed Res, Biopathol & Regenerat Med Inst IBIMER, E-18071 Granada, Spain, [Marchal, Juan A.] Univ Granada, Fac Med, Dept Human Anat & Embryol, E-18016 Granada, Spain, [Marchal, Juan A.] Univ Granada, Univ Hosp Granada, Biosanitary Inst Granada Ibs GRANADA, E-18016 Granada, Spain, [Montanez-Heredia, Elvira] Inst Invest Biomed Malaga IBIMA, E-29010 Malaga, Spain, [Estades-Rubio, JF, García-Piriz,M, García-Vera,J, Montañez-Heredia,E] Department of Orthopedic Surgery and Traumatology, Virgen de la Victoria University Hospital, Málaga, Spain. [Reyes-Martín,A] Department of Orthopedic Surgery and Traumatology, Hospital Regional Universitario Carlos Haya, Malaga, Málaga, Spain. [Morales-Marcos,V] Department of Orthopedic Surgery and Traumatology, Hospital Quirónsalud Málaga, Málaga, Spain. [Perán,M] Department of Health Sciences, University of Jaén, Jaén, Spain. [Perán,M, and Marchal,JA] Biopathology and Regenerative Medicine Institute (IBIMER), Centre for Biomedical Research, University of Granada, Granada, Spain. [Marchal,JA] Department of Human Anatomy and Embryology, Faculty of Medicine, University of Granada, Granada, Spain. Biosanitary Institute of Granada (ibs. GRANADA), University Hospitals of Granada-University of Granada, Granada, Granada, Spain. [Montañez-Heredia,E] Instituto de Investigación Biomédica de Málaga (IBIMA), Málaga, Spain.

Subjects
Replacement, Arthritis, Osteoarthritis, Stabilized hyaluronic acid (NASHA), lcsh:Chemistry, Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings], Analytical, Diagnostic and Therapeutic Techniques and Equipment::Therapeutics::Drug Therapy::Drug Administration Routes::Injections::Injections, Intra-Articular [Medical Subject Headings], chemistry.chemical_compound, 0302 clinical medicine, hyaluronic acid, Hyaluronic acid, Inyecciones intraarticulares, Prevalence, Diseases::Musculoskeletal Diseases::Joint Diseases::Arthritis::Osteoarthritis::Osteoarthritis, Knee [Medical Subject Headings], Medicine, 030212 general & internal medicine, Masculino, Costos de la atención en salud, lcsh:QH301-705.5, Spectroscopy, Persons::Persons::Age Groups::Adult::Aged [Medical Subject Headings], Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Methods::Research Design::Random Allocation [Medical Subject Headings], Adulto, Femenino, General Medicine, Cost-effectiveness analysis, Metaanalysis, Humanos, Management, Computer Science Applications, Anesthesia, Distribución aleatoria, Health Care::Health Care Quality, Access, and Evaluation::Delivery of Health Care::Health Care Costs [Medical Subject Headings], Knee osteoarthritis, medicine.medical_specialty, Ácido hialurónico, WOMAC, Efficacy, Anciano, Check Tags::Male [Medical Subject Headings], Chemicals and Drugs::Carbohydrates::Polysaccharides::Glycosaminoglycans::Hyaluronic Acid [Medical Subject Headings], Burden, Vial, Article, knee osteoarthritis, Catalysis, Inorganic Chemistry, 03 medical and health sciences, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Therapeutics::Drug Therapy::Drug Administration Routes::Injections::Injections, Intra-Articular::Viscosupplementation [Medical Subject Headings], Physical and Theoretical Chemistry, Persons::Persons::Age Groups::Adult [Medical Subject Headings], Molecular Biology, 030203 arthritis & rheumatology, Mediana edad, Hip, business.industry, Organic Chemistry, Hyalunonic acid, Persons::Persons::Age Groups::Adult::Middle Aged [Medical Subject Headings], Single injection, medicine.disease, Surgery, lcsh:Biology (General), lcsh:QD1-999, Check Tags::Female [Medical Subject Headings], chemistry, Osteoartritis de la rodilla, Viscosupplementation, business, Viscosuplementación
Abstract

Given the wide difference in price per vial between various presentations of hyaluronic acid, this study seeks to compare the effectiveness and treatment cost of stabilized hyaluronic acid (NASHA) in a single injection with standard preparations of hyaluronic acid (HA) in five injections in osteoarthritis (OA) of the knee. Fifty-four patients with knee osteoarthritis (Kellgren-Lawrence Grade II and III) and the Western Ontario and McMaster Universities Arthritis Index (WOMAC) pain score greater than 7, with a homogeneous distribution of age, sex, BMI, and duration of disease, were included in this study. Patients were randomized into two groups: Group I was treated with NASHA (Durolane(®)) and Group II with HA (Go-ON(®)). Patient's evolution was followed up at the 1st, 2nd, 4th, 8th, 12th, and 26th week after treatment. A statistically significant improvement in WOMAC score was observed for patients treated with NASHA versus those who received HA at Week 26. In addition, the need for analgesia was significantly reduced at Week 26 in the NASHA-treated group. Finally, the economic analysis showed an increased cost of overall treatment with HA injections. Our data support the use of the NASHA class of products in the treatment of knee OA. Yes

Published
2017

24. Design, synthesis, HER2 inhibition and anticancer evaluation of new substituted 1,5-dihydro-4,1-benzoxazepines

Author

Matilde Ner, Juan A. Marchal, Yaiza Jiménez-Martínez, Joaquín M. Campos, Hugo Gutiérrez-de-Terán, Ana Conejo-García, Houria Boulaiz, Francho Nerín-Fonz, David Araripe, Olga Cruz-Lopez, [Cruz-López,O, Ner,M, Campos,JM, Conejo-García,A] Department of Medicinal and Organic Chemistry, Faculty of Pharmacy, University of Granada, Granada, Spain. [Cruz-López,O, Jiménez-Martínez,Y, Marchal,JA, Boulaiz,H, Conejo-García,A] Biosanitary Institute of Granada (ibs.GRANADA), SAS-University of Granada, Granada, Spain. [Nerín-Fonz,F, Araripe,D, Gutiérrez-de-Terán,H] Department of Cell and Molecular Biology, Uppsala University, Uppsala, Sweeden. [Jiménez-Martínez,Y, Boulaiz,H] Biopathology and Medicine Regenerative Institute, University of Granada, Granada, Spain. [Jiménez-Martínez,Y, Boulaiz,H] Excellence Research Unit 'Modeling Nature' (MNat), Department of Human Anatomy and Embryology, University of Granada, Granada, Spain., and A. C. -G. is thankful to Consejería de Economía, Conocimiento, Empresas y Universidad of the Junta de Andalucía (Excellence Research Project P18-RT-1679) and the Oficina de Transferencia de Resultados de Investigación of the University of Granada (PR/17/006 project) for financial support. J. A. M. and H. B. thanks Instituto de Salud Carlos III (RTI2018-101309-B-C22), Fundación Mutua Madrileña (project FMM-AP16683-2017), Consejería de Salud Junta de Andalucía (PI-0089-2017) for financial support.

Subjects
Anatomy::Cells::Cells, Cultured::Cell Line::Cell Line, Tumor [Medical Subject Headings], Models, Molecular, Chemicals and Drugs::Chemical Actions and Uses::Pharmacologic Actions::Molecular Mechanisms of Pharmacological Action::Enzyme Inhibitors::Protein Kinase Inhibitors [Medical Subject Headings], Receptor, ErbB-2, Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Hydrolases::Peptide Hydrolases::Cysteine Proteases::Cysteine Endopeptidases::Caspases::Caspases, Effector::Caspase 3 [Medical Subject Headings], Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings], Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Transferases::Phosphotransferases::Phosphotransferases (Alcohol Group Acceptor)::Protein Kinases::Protein-Tyrosine Kinases::Receptor Protein-Tyrosine Kinases::Receptor, erbB-2 [Medical Subject Headings], Cancer cell line, Modelos moleculares, Drug Discovery, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Models, Theoretical::Models, Molecular [Medical Subject Headings], Molecular Structure, pyroptosis, Biochemistry and Molecular Biology, Läkemedelskemi, General Medicine, molecular modelling, Phenomena and Processes::Chemical Phenomena::Biochemical Phenomena::Structure-Activity Relationship [Medical Subject Headings], Benzoxazepines, Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Hydrolases::Peptide Hydrolases::Cysteine Proteases::Cysteine Endopeptidases::Caspases::Caspases, Initiator::Caspase 1 [Medical Subject Headings], Molecular modelling, Piroptosis, Receptor, Research Article, Research Paper, HER2 receptor, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Diagnosis::Diagnostic Techniques and Procedures::Clinical Laboratory Techniques::Cytological Techniques::Drug Screening Assays, Antitumor [Medical Subject Headings], Antineoplastic Agents, RM1-950, benzoxazepines, Structure-Activity Relationship, Phenomena and Processes::Chemical Phenomena::Molecular Structure [Medical Subject Headings], Political science, HER2, Cell Line, Tumor, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Toxicity Tests::Inhibitory Concentration 50 [Medical Subject Headings], Pyroptosis, Humans, Her2 receptor, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Drug Discovery::Drug Design [Medical Subject Headings], Phenomena and Processes::Cell Physiological Phenomena::Cell Physiological Processes::Cell Growth Processes::Cell Proliferation [Medical Subject Headings], Protein Kinase Inhibitors, Antitumour, Cell Proliferation, Pharmacology, Genes erbB-2, Línea celular tumoral, Dose-Response Relationship, Drug, Phenomena and Processes::Physiological Phenomena::Pharmacological Phenomena::Dose-Response Relationship, Drug [Medical Subject Headings], Chemicals and Drugs::Chemical Actions and Uses::Pharmacologic Actions::Therapeutic Uses::Antineoplastic Agents [Medical Subject Headings], Antineoplásicos, Design synthesis, Drug Design, Therapeutics. Pharmacology, Medicinal Chemistry, Drug Screening Assays, Antitumor, Humanities, Biokemi och molekylärbiologi
Abstract

A. C. -G. is thankful to Consejeria de Economia, Conocimiento, Empresas y Universidad of the Junta de Andalucia (Excellence Research Project P18-RT-1679) and the Oficina de Transferencia de Resultados de Investigacion of the University of Granada (PR/17/006 project) for financial support. J. A. M. and H. B. thanks Instituto de Salud Carlos III (RTI2018-101309-B-C22), Fundacion Mutua Madrilena (project FMM-AP16683-2017), Consejeria de Salud Junta de Andalucia (PI-0089-2017) for financial support., A series of 11 new substituted 1,5-dihydro-4,1-benzoxazepine derivatives was synthesised to study the influence of the methyl group in the 1-(benzenesulphonyl) moiety, the replacement of the purine by the benzotriazole bioisosteric analogue, and the introduction of a bulky substituent at position 6 of the purine, on the biological effects. Their inhibition against isolated HER2 was studied and the structure–activity relationships have been confirmed by molecular modelling studies. The most potent compound against isolated HER2 is 9a with an IC50 of 7.31 mM. We have investigated the effects of the target compounds on cell proliferation. The most active compound (7c) against all the tumour cell lines studied (IC50 0.42–0.86 mM) does not produce any modification in the expression of pro-caspase 3, but increases the caspase 1 expression, and promotes pyroptosis., Junta de Andalucia P18-RT-1679, Oficina de Transferencia de Resultados de Investigacion of the University of Granada PR/17/006, Instituto de Salud Carlos III European Commission RTI2018-101309-B-C22, Instituto de Salud Carlos III FMM-AP16683-2017, Junta de Andalucia PI-0089-2017

Published
2021

25. How Can Nanotechnology Help to Repair the Body? Advances in Cardiac, Skin, Bone, Cartilage and Nerve Tissue Regeneration

Author

Perán, Macarena, Garcia, Maria Angel, Lopez-Ruiz, Elena, Jiménez, Gema, Marchal, Juan Antonio, [Perán,M, Lopez-Ruiz,E] Department of Health Sciences, University of Jaén, Jaén, Spain. [Garcia, MA] Research Unit, University Hospital 'Virgen de las Nieves', Granada, Spain. [Jiménez,G, Marchal,JA] Biopathology and Regenerative Medicine Institute (IBIMER), University of Granada, Armilla, Granada, Spain. [Marchal, JA] Department of Human Anatomy and Embryology, University of Granada, Granada, Spain., and This work was supported in part by grants from the Consejería de Economía, Innovación y Ciencia (Junta de Andalucía, excellence project number CTS-6568) and the Instituto de Salud Carlos III (Fondo de Investigación Sanitaria, FEDER funds, grant number PI10/02295).

Subjects
bio-scaffold, Anatomy::Musculoskeletal System::Skeleton::Bone and Bones [Medical Subject Headings], Analytical, Diagnostic and Therapeutic Techniques and Equipment::Equipment and Supplies::Culture Media::Tissue Scaffolds [Medical Subject Headings], nanotechnology, Nanoestructuras, Anatomy::Musculoskeletal System::Cartilage [Medical Subject Headings], Andamios del Tejido, Disciplines and Occupations::Natural Science Disciplines::Nanotechnology [Medical Subject Headings], Anatomy::Integumentary System::Skin [Medical Subject Headings], Anatomy::Tissues::Nerve Tissue [Medical Subject Headings], tissue engineering, Ingeniería de Tejidos, nanostructures, Anatomy::Cardiovascular System::Heart [Medical Subject Headings], Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Culture Techniques::Cell Engineering::Tissue Engineering [Medical Subject Headings], Technology, Industry, Agriculture::Technology, Industry, and Agriculture::Manufactured Materials::Nanostructures [Medical Subject Headings]
Abstract

(This article belongs to the Special Issue Advances in Nanoscale Biomaterials) Nanotechnologists have become involved in regenerative medicine via creation of biomaterials and nanostructures with potential clinical implications. Their aim is to develop systems that can mimic, reinforce or even create in vivo tissue repair strategies. In fact, in the last decade, important advances in the field of tissue engineering, cell therapy and cell delivery have already been achieved. In this review, we will delve into the latest research advances and discuss whether cell and/or tissue repair devices are a possibility. Focusing on the application of nanotechnology in tissue engineering research, this review highlights recent advances in the application of nano-engineered scaffolds designed to replace or restore the followed tissues: (i) skin; (ii) cartilage; (iii) bone; (iv) nerve; and (v) cardiac. Yes

Published
2013

26. Functionalized nanostructures with application in regenerative medicine

Author

Macarena Perán, Milán Bustamante, Elena López-Ruiz, Juan A. Marchal, Roberto Madeddu, Gema Jiménez, María Ángel García, [Perán,M, López-Ruiz,E] Department of Health Sciences, University of Jaén, Jaén, Spain. [García,MA] Research Unit, Hospital Universitario Virgen de las Nieves, Granada, Spain. [Bustamante,M] Biosciences Institute, University College Cork, Cork, Ireland. [Jiménez,G, Marchal,JA] Biopathology and Regenerative Medicine Institute (IBIMER), Biomedical Research Centre, University of Granada, Granada, Spain. [Madeddu,R] Department of Biomedical Sciences, University of Sassari, Sassari, Italy. [Marchal,JA] Department of Human Anatomy and Embryology, Faculty of Medicine, University of Granada, Granada, Spain., and This work was supported in part by grants from the Consejería de Economía, Innovación y Ciencia (Junta de Andalucía, excellence project number CTS-6568) and the Instituto de Salud Carlos III (Fondo de Investigación Sanitaria, FEDER funds, grant number PI10/02295).

Subjects
Analytical, Diagnostic and Therapeutic Techniques and Equipment::Equipment and Supplies::Culture Media::Tissue Scaffolds [Medical Subject Headings], Bone Regeneration, Cell- and Tissue-Based Therapy, Biocompatible Materials, Regeneración Ósea, Review, Stem cells, Regenerative medicine, bone, Cell therapy, lcsh:Chemistry, Phenomena and Processes::Musculoskeletal and Neural Physiological Phenomena::Musculoskeletal Physiological Phenomena::Musculoskeletal Physiological Processes::Bone Remodeling::Bone Regeneration [Medical Subject Headings], Drug Delivery Systems, Tissue engineering, Medicine, Nanotechnology, Cell encapsulation, cartilage, gene and drug delivery, lcsh:QH301-705.5, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Culture Techniques::Cell Engineering::Tissue Engineering [Medical Subject Headings], Spectroscopy, nanomaterials, Stem cell, Tissue Scaffolds, Gene Transfer Techniques, Nanopartículas, Técnicas de Transferencia de Gen, General Medicine, Andamios del Tejido, Anatomy::Tissues::Connective Tissue::Cartilage [Medical Subject Headings], BIO/17 Istologia, Computer Science Applications, Cell tracking, Sistemas de Liberación de Medicamentos, cell tracking, Disciplines and Occupations::Health Occupations::Medicine::Regenerative Medicine [Medical Subject Headings], scaffolds, Technology, Industry, Agriculture::Technology, Industry, and Agriculture::Manufactured Materials::Nanostructures::Nanoparticles [Medical Subject Headings], Cartílago, regenerative medicine, Anatomy::Cells::Stem Cells::Pluripotent Stem Cells [Medical Subject Headings], Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Genetic Techniques::Gene Transfer Techniques [Medical Subject Headings], Gene delivery, cell encapsulation, Catalysis, Inorganic Chemistry, stem cells, Ingeniería de Tejidos, Humans, Physical and Theoretical Chemistry, Bone regeneration, Bone, Molecular Biology, Technology, Industry, Agriculture::Technology, Industry, and Agriculture::Manufactured Materials::Nanostructures [Medical Subject Headings], Nanomaterials, Scaffolds, Tissue Engineering, business.industry, Organic Chemistry, Nanoestructuras, Medicina Regenerativa, Nanostructures, Transplantation, Cartilage, lcsh:Biology (General), lcsh:QD1-999, Gene and drug delivery, cell therapy, business, Células Madre Pluripotentes, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Therapeutics::Drug Therapy::Drug Delivery Systems [Medical Subject Headings], Stem Cell Transplantation
Abstract

In the last decade, both regenerative medicine and nanotechnology have been broadly developed leading important advances in biomedical research as well as in clinical practice. The manipulation on the molecular level and the use of several functionalized nanoscaled materials has application in various fields of regenerative medicine including tissue engineering, cell therapy, diagnosis and drug and gene delivery. The themes covered in this review include nanoparticle systems for tracking transplanted stem cells, self-assembling peptides, nanoparticles for gene delivery into stem cells and biomimetic scaffolds useful for 2D and 3D tissue cell cultures, transplantation and clinical application., Consejería de Economía, Innovación y Ciencia, Junta de Andalucía CTS-6568, Instituto de Salud Carlos III, Fondo de Investigación Sanitaria, FEDER PI10/02295

Published
2012

27. Large-Scale Production of Lentiviral Vectors: Current Perspectives and Challenges

Author

Martínez-Molina, Eduardo, Chocarro-Wrona, Carlos, Martínez-Moreno, Daniel, Marchal, Juan A., Boulaiz, Houria, [Martínez-Molina,E, Chocarro-Wrona,C, Martínez-Moreno,D, Marchal,JA, Boulaiz,H] Biopathology and Medicine Regenerative Institute (IBIMER), University of Granada (D.M.), Granada, Spain. [Chocarro-Wrona,C, Boulaiz,H] Department of Human Anatomy and Embryology, University of Granada, Granada, Spain. [Chocarro-Wrona,C, Boulaiz,H] Excellence Research Unit 'Modeling Nature' (MNat), University of Granada, Granada, Spain. [Chocarro-Wrona,C, Boulaiz,H] Biosanitary Institute of Granada (ibs.GRANADA), SAS-Universidad de Granada, Granada, Spain., and This research was supported by Fundacion Empresa Universidad de Granada (Project PR/18/001), Fundación Mutua Madrileña (project FMM-AP16683-2017), Consejería de Salud Junta de Andalucía (PI-0089-2017), from the Chair 'Doctors Galera-Requena in cancer stem cell research' and Consejería de Economía, Conocimiento, Empresas y Universidades de la Junta de Andalucía (SOMM17/6109/UGR, FEDER Funds).

Subjects
Bioreactor, Anatomy::Cells [Medical Subject Headings], Organisms::Viruses::RNA Viruses::Retroviridae::Lentivirus [Medical Subject Headings], Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Genetic Techniques::Gene Transfer Techniques [Medical Subject Headings], Large-scale production, Terapia genética, Gene therapy, Anatomy::Cells::Cells, Cultured::Cell Line [Medical Subject Headings], Analytical, Diagnostic and Therapeutic Techniques and Equipment::Equipment and Supplies::Bioreactors [Medical Subject Headings], Information Science::Information Science::Communications Media::Publications::Review Literature as Topic [Medical Subject Headings], Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Clinical Laboratory Techniques::Cytological Techniques::Cell Culture Techniques [Medical Subject Headings], Lentiviral vector, Reactores biológicos, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Therapeutics::Biological Therapy::Genetic Therapy [Medical Subject Headings]
Abstract

Lentiviral vectors (LVs) have gained value over recent years as gene carriers in gene therapy. These viral vectors are safer than what was previously being used for gene transfer and are capable of infecting both dividing and nondividing cells with a long-term expression. This characteristic makes LVs ideal for clinical research, as has been demonstrated with the approval of lentivirus-based gene therapies from the Food and Drug Administration and the European Agency for Medicine. A large number of functional lentiviral particles are required for clinical trials, and large-scale production has been challenging. Therefore, efforts are focused on solving the drawbacks associated with the production and purification of LVsunder current good manufacturing practice. In recent years, we have witnessed the development and optimization of new protocols, packaging cell lines, and culture devices that are very close to reaching the target production level. Here, we review the most recent, efficient, and promising methods for the clinical-scale production ofLVs. Yes

Published
2020

29. Immune cell impact of three differently coated lipid nanocapsules: pluronic, chitosan and polyethylene glycol

Author

Francesco Sgarrella, Cristiano Farace, Lucia Gemma Delogu, Marco Orecchioni, Juan A. Marchal, J. M. Peula-Garcia, Yolande Asara, Roberto Madeddu, Paola Sánchez-Moreno, Roberto Manetti, [Farace,C, Asara,Y, Madeddu,R] Department of Biomedical Science, University of Sassari, Sassari, Italy. [Sánchez-Moreno,P, Peula-García,JM, Delogu,LG] Biocolloid and Fluid Physics Group, Department of Applied Physics, University of Granada, Granada, Spain. [Orecchioni,M, Manetti,R, Sgarrella,F] Department of Chemistry and Pharmacy University of Sassari, Sassari, Italy. [Peula-García,JM] Department of Applied Physics II, University of Málaga,Málaga, Spain. [Marchal,JA] Biopathology and Medicine Regenerative Institute (IBIMER), University of Granada, Granada, Spain. Biosanitary Institute of Granada (ibs.GRANADA), University Hospitals of Granada-Univesity of Granada, Granada, Spain. Department of Human Anatomy and Embryology, University of Granada, Granada, Spain. [Madeddu,R]National Institute of Biostructures and Biosystem (INBB), Rome, Italy., and This work was partly supported by the Fondazione Banco di Sardegna (grant N° 2013.1308, 2014.6035 to L.G.D.), the University of Sassari (Italy), the Sardinia Region (grant N° CRP-59720 to L.G.D.), the Gianfranco del Prete Association 'The future: medicine, biology and nanotechnology Award' to L.G.D, the Instituto de Salud Carlos III (FEDER funds, grant N° PI10/02295 to J.A.M), the Consejería de Economía, Innovación y Ciencia (Junta de Andalucía, excellence grant N° (CTS-6568 to J.A.M) and by the grant N° (MAT2013-43922-R).

Subjects
Biocompatible, Polímeros, Polietilenglicoles, Erythrocytes, T-Lymphocytes, medicine.medical_treatment, Chemicals and Drugs::Organic Chemicals::Alcohols::Glycols::Ethylene Glycols::Polyethylene Glycols::Poloxamer [Medical Subject Headings], 02 engineering and technology, Linfocitos t, Leucocitos mononucleares, Lymphocyte Activation, 01 natural sciences, Monocytes, Polyethylene Glycols, Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings], Coated Materials, Biocompatible, Anatomy::Cells::Blood Cells::Leukocytes::Leukocytes, Mononuclear::Monocytes [Medical Subject Headings], Inmunomodulación, Leukocytes, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Therapeutics::Biological Therapy::Immunomodulation::Immunotherapy [Medical Subject Headings], Inmunidad innata, Ácido láctico, Poloxámero, Anatomy::Cells::Blood Cells::Leukocytes::Leukocytes, Mononuclear::Lymphocytes::T-Lymphocytes [Medical Subject Headings], Multidisciplinary, Chemistry, Citocinas, Nanocápsulas, 021001 nanoscience & nanotechnology, Lipids, Humanos, Quitosano, Cytokine, medicine.anatomical_structure, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Diagnosis::Diagnostic Techniques and Procedures::Clinical Laboratory Techniques::Cytological Techniques::Cell Separation::Flow Cytometry [Medical Subject Headings], Lípidos, Cytokines, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Therapeutics::Biological Therapy::Immunomodulation [Medical Subject Headings], 0210 nano-technology, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Diagnosis::Diagnostic Techniques and Procedures::Diagnostic Imaging::Microscopy::Microscopy, Confocal [Medical Subject Headings], Monocitos, Biocompatibility, Cell Survival, Mononuclear, Cell Proliferation, Humans, Immune System, Leukocytes, Mononuclear, Chitosan, Nanocapsules, Chemicals and Drugs::Pharmaceutical Preparations::Dosage Forms::Drug Carriers::Nanocapsules [Medical Subject Headings], Phenomena and Processes::Cell Physiological Phenomena::Cell Physiological Processes::Cell Death::Necrosis [Medical Subject Headings], Chemicals and Drugs::Lipids [Medical Subject Headings], 010402 general chemistry, Peripheral blood mononuclear cell, Article, Immune system, medicine, Phenomena and Processes::Immune System Phenomena::Immunity::Immunity, Innate [Medical Subject Headings], Inmunoterapia, Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Peptides::Intercellular Signaling Peptides and Proteins::Cytokines [Medical Subject Headings], Microscopía confocal, Chemicals and Drugs::Macromolecular Substances::Polymers::Biopolymers::Chitin::Chitosan [Medical Subject Headings], Innate immune system, Monocyte, Anatomy::Cells::Blood Cells::Leukocytes::Leukocytes, Mononuclear [Medical Subject Headings], Coated Materials, Chemicals and Drugs::Organic Chemicals::Carboxylic Acids::Hydroxy Acids::Lactates::Lactic Acid [Medical Subject Headings], Immunotherapy, Chemicals and Drugs::Macromolecular Substances::Polymers [Medical Subject Headings], Phenomena and Processes::Cell Physiological Phenomena::Cell Physiological Processes::Cell Death::Apoptosis [Medical Subject Headings], 0104 chemical sciences, Immunology, Biophysics, Chemicals and Drugs::Organic Chemicals::Alcohols::Glycols::Ethylene Glycols::Polyethylene Glycols [Medical Subject Headings], Citometría de flujo
Abstract

Lipid nanocapsules (NCs) represent promising tools in clinical practice for diagnosis and therapy applications. However, the NC appropriate functionalization is essential to guarantee high biocompatibility and molecule loading ability. In any medical application, the immune system-impact of differently functionalized NCs still remains to be fully understood. A comprehensive study on the action exerted on human peripheral blood mononuclear cells (PBMCs) and major immune subpopulations by three different NC coatings: pluronic, chitosan and polyethylene glycol-polylactic acid (PEG) is reported. After a deep particle characterization, the uptake was assessed by flow-cytometry and confocal microscopy, focusing then on apoptosis, necrosis and proliferation impact in T cells and monocytes. Cell functionality by cell diameter variations, different activation marker analysis and cytokine assays were performed. We demonstrated that the NCs impact on the immune cell response is strongly correlated to their coating. Pluronic-NCs were able to induce immunomodulation of innate immunity inducing monocyte activations. Immunomodulation was observed in monocytes and T lymphocytes treated with Chitosan-NCs. Conversely, PEG-NCs were completely inert. These findings are of particular value towards a pre-selection of specific NC coatings depending on biomedical purposes for pre-clinical investigations; i.e. the immune-specific action of particular NC coating can be excellent for immunotherapy applications.

Published
2016
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30. Activin A/BMP2 chimera AB235 drives efficient redifferentiation of long term cultured autologous chondrocytes

Author

Esmeralda Carrillo, Gema Jiménez, Witek Kwiatkowski, Elvira Montañez, J. C. Izpisua Belmonte, Macarena Perán, Peter C. Gray, Juan A. Marchal, Senyon Choe, Francisco Arrebola, Elena López-Ruiz, [Jiménez,G, Carrillo,E, Marchal,JA] Biopathology and Regenerative Medicine Institute (IBIMER), Centre for Biomedical Research, University of Granada, Granada, Spain. Department of Human Anatomy and Embryology, Faculty of Medicine, University of Granada, Granada, Spain. Biosanitary Institute of Granada (ibs.GRANADA), University Hospitals of Granada-Univesity of Granada, Granada, Spain. [López-Ruiz,E, Perán,M] Department of Health Sciences, University of Jaén, Jaén, Spain. [Kwiatkowski,W, Choe,S] Structural Biology Laboratory, Salk Institute for Biological Studies, La Jolla, California, USA. [Montañez,E] Department of Orthopedic Surgery and Traumatology, Virgen de la Victoria University Hospital, Málaga, Spain. [Arrebola,F] Department of Histology, Faculty of Medicine, University of Granada, Granada, Spain. [Gray,PC] Clayton Foundation Laboratories for Peptide Biology, Salk Institute for Biological Studies, La Jolla CA, California, USA. [Izpisua Belmonte, JC] Gene Expression Laboratory, Salk Institute for Biological Studies, La Jolla CA, California, USA. [Choe,S] Qualcomm Institute, Univ. California, San Diego, La Jolla, USA., and This work was supported by the Consejería de Economía, Innovación y Ciencia (Junta de Andalucía, excellence project number CTS-6568).

Subjects
Cartilage, Articular, Male, Time Factors, Cellular differentiation, Cell, Diseases::Musculoskeletal Diseases::Rheumatic Diseases::Osteoarthritis [Medical Subject Headings], Chemicals and Drugs::Hormones, Hormone Substitutes, and Hormone Antagonists::Hormones::Gonadal Hormones::Activins [Medical Subject Headings], Bone Morphogenetic Protein 2, Gene Expression, Anatomy::Cells::Connective Tissue Cells::Chondrocytes [Medical Subject Headings], Anatomy::Tissues::Connective Tissue::Subcutaneous Tissue [Medical Subject Headings], Mice, SCID, Ligands, Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings], Extracellular matrix, Mice, Mice, Inbred NOD, Organisms::Eukaryota::Animals [Medical Subject Headings], Activinas, Autologous chondrocyte implantation, Tejido subcutáneo, Anatomy::Cells::Cellular Structures::Extracellular Space::Extracellular Matrix [Medical Subject Headings], Multidisciplinary, Reverse Transcriptase Polymerase Chain Reaction, Cell Differentiation, Middle Aged, Immunohistochemistry, Humanos, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Genetic Techniques::Nucleic Acid Amplification Techniques::Polymerase Chain Reaction::Reverse Transcriptase Polymerase Chain Reaction [Medical Subject Headings], Activins, Extracellular Matrix, Cell biology, Chemicals and Drugs::Carbohydrates::Polysaccharides::Proteoglycans [Medical Subject Headings], medicine.anatomical_structure, Anatomy::Musculoskeletal System::Cartilage::Hyaline Cartilage::Cartilage, Articular [Medical Subject Headings], Female, Proteoglycans, Matriz extracelular, Collagen, Cartílago articular, Proteoglicanos, Transplantation, Heterologous, Biology, Transplantation, Autologous, Bone morphogenetic protein 2, Article, Chondrocytes, In vivo, Osteoarthritis, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Diagnosis::Diagnostic Techniques and Procedures::Clinical Laboratory Techniques::Cytological Techniques::Cytodiagnosis::Biopsy [Medical Subject Headings], medicine, Animals, Humans, Aged, Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Rodentia::Muridae::Murinae::Mice [Medical Subject Headings], Reacción en cadena de la polimerasa de transcriptasa inversa, Chemicals and Drugs::Macromolecular Substances::Polymers::Biopolymers::Collagen [Medical Subject Headings], Chondrogenesis, Transplantation, Biopsia, Condrocitos, Osteoartritis, Animales, Immunology, Colágeno
Abstract

Autologous chondrocyte implantation (ACI) depends on the quality and quantity of implanted cells and is hindered by the fact that chondrocytes cultured for long periods of time undergo dedifferentiation. Here we have developed a reproducible and efficient chondrogenic protocol to redifferentiate chondrocytes isolated from osteoarthritis (OA) patients. We used morphological, histological and immunological analysis together with a RT-PCR detection of collagen I and collagen II gene expression to show that chondrocytes isolated from articular cartilage biopsies of patients and subjected to long-term culture undergo dedifferentiation and that these cells can be redifferentiated following treatment with the chimeric Activin A/BMP2 ligand AB235. Examination of AB235-treated cell pellets in both in vitro and in vivo experiments revealed that redifferentiated chondrocytes synthesized a cartilage-specific extracellular matrix (ECM), primarily consisting of vertically-orientated collagen fibres and cartilage-specific proteoglycans. AB235-treated cell pellets also integrated into the surrounding subcutaneous tissue following transplantation in mice as demonstrated by their dramatic increase in size while non-treated control pellets disintegrated upon transplantation. Thus, our findings describe an effective protocol for the promotion of redifferentiation of autologous chondrocytes obtained from OA patients and the formation of a cartilage-like ECM that can integrate into the surrounding tissue in vivo.

Published
2015

31. EMT and EGFR in CTCs cytokeratin negative non-metastatic breast cancer

Author

Serrano, Maria J., Ortega, Francisco G., Alvarez-Cubero, Maria J., Nadal Rios, Rosa, Sanchez-Rovira, Pedro, Salido, Marta, Rodríguez, María, García-Puche, José Luis, Delgado-Rodríguez, Miguel, Solé, Francisco, García, Maria A., Perán, Macarena, Rosell, Rafael, Marchal, Juan A., Lorente, Jose A., Universitat Autònoma de Barcelona, [Serrano,MJ, Ortega,FG, Alvarez-Cubero,MJ, García-Puche,JL, Lorente,JA] GENYO. Pfizer-University of Granada-Andalusian Government Centre of Genomics and Oncology, Granada, Spain. [Serrano,MJ, Lorente,JA] Laboratory of Genetic Identification-UGR, Department of Legal Medicine, University of Granada, Granada, Spain. [Nadal,R] Hospital de Barcelona, Medical Oncology Department, Barcelona, Spain. [Sanchez-Rovira,P, Delgado-Rodriguez,M, Perán,M] University of Jaén, Division of Preventive Medicine and Public Health, CIBERESP, Jaén, Spain. [Salido,M, Rodríguez,M] Molecular Cytogenetics Laboratory, Pathology Department, Parc de Salut Mar-Hospital del Mar-IMIM-GRETNHE, Barcelona, Spain. [Solé,F] Medicine Department. Universitat Autònoma de Barcelona, Barcelona, Spain, Institut de Recerca contra la Leucèmia Josep Carreras, Badalona, Spain. [García,MA] Department of Oncology, Virgen de las Nieves, University Hospital, Granada, Spain. [Rosell,R] Catalan Institute of Oncology, Hospital Germans Trias i Pujol, Badalona, Spain (RR), Pangaea Biotech SL, USP Dexeus University Institute, Barcelona, Spain (RR, MAM). [Perán,M, Marchal,JA] Biopathology and Regenerative Medicine Institute (IBIMER), Centre for Biomedical Research, University of Granada, Granada, Spain. [Marchal,JA] Department of Human Anatomy and Embryology, University of Granada, Granada, Spain., and Public Health and Progress Foundation. Ministry of Health, Andalusian Government and the Instituto de Salud Carlos III (PI10/02295, FEDER funds)

Subjects
Oncology, Anatomy::Cells::Cells, Cultured::Cell Line::Cell Line, Tumor [Medical Subject Headings], Pathology, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Microscopy::Microscopy, Confocal [Medical Subject Headings], Phenomena and Processes::Genetic Phenomena::Phenotype [Medical Subject Headings], Fluorescent Antibody Technique, Apoptosis, Named Groups::Persons::Age Groups::Adult::Middle Aged [Medical Subject Headings], void(null), al Subject Headings::Chemicals and Drugs::Macromolecular Substances::Polymers::Biopolymers::Intermediate Filament Proteins::Vimentin [Medicjavascript], Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings], Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Models, Theoretical [Medical Subject Headings], Circulating tumor cell, Vimentina, Organisms::Eukaryota::Animals [Medical Subject Headings], Anatomy::Cells::Cellular Structures::Intracellular Space::Cytoplasm::Cytoplasmic Structures::Cytoskeleton::Intermediate Filaments [Medical Subject Headings], Anatomy::Cells::Neoplastic Cells, Circulating [Medical Subject Headings], Microscopy, Confocal, Transición epitelial-mesenquimal, Pronóstico, Marcadores biológicos de tumor, Middle Aged, University hospital, Neoplastic Cells, Circulating, Slug, Humanos, ErbB Receptors, Queratinas, Neoplasias de la mama, embryonic structures, MCF-7 Cells, Keratins, Female, Modelos teóricos, Chemicals and Drugs::Biological Factors::Biological Markers::Tumor Markers, Biological [Medical Subject Headings], Fenotipo, Research Paper, Diseases::Neoplasms::Neoplastic Processes::Neoplasm Metastasis::Neoplastic Cells, Circulating [Medical Subject Headings], Adult, medicine.medical_specialty, Organisms::Eukaryota::Animals::Invertebrates::Mollusca::Gastropoda [Medical Subject Headings], Epithelial-Mesenchymal Transition, Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Cytoskeletal Proteins::Intermediate Filament Proteins::Vimentin [Medical Subject Headings], EGFR, Blotting, Western, Breast Neoplasms, Filamentos intermedios, Biology, Real-Time Polymerase Chain Reaction, Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Transcription Factors [Medical Subject Headings], Cytokeratin, Breast cancer, Internal medicine, Breast Cancer, medicine, Biomarkers, Tumor, Non metastatic, Humans, Vimentin, In patient, Chemicals and Drugs::Macromolecular Substances::Polymers::Biopolymers::Intermediate Filament Proteins::Keratins [Medical Subject Headings], Bcl-2, Egfr signaling, Diseases::Neoplasms::Neoplasms by Site::Breast Neoplasms [Medical Subject Headings], Aged, Línea celular tumoral, Experimental model, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Genetic Techniques::Nucleic Acid Amplification Techniques::Polymerase Chain Reaction::Real-Time Polymerase Chain Reaction [Medical Subject Headings], Gastrópodos, Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Membrane Proteins::Receptors, Cell Surface::Receptors, Peptide::Receptors, Growth Factor::Receptor, Epidermal Growth Factor [Medical Subject Headings], medicine.disease, Animales, Phenomena and Processes::Cell Physiological Phenomena::Cell Physiological Processes::Cell Transdifferentiation::Epithelial-Mesenchymal Transition [Medical Subject Headings], Analytical, Diagnostic and Therapeutic Techniques and Equipment::Diagnosis::Prognosis [Medical Subject Headings], Snail Family Transcription Factors, Células neoplásicas circulantes, Circulating Tumor Cells, Transcription Factors
Abstract

// Maria J. Serrano 1,2,* , Francisco G. Ortega 1,* , Maria J. Alvarez-Cubero 1,2 , Rosa Nadal 3 , Pedro Sanchez-Rovira 4 , Marta Salido 5 , Maria Rodriguez 5 , Jose L. Garcia-Puche 1 , Miguel Delgado-Rodriguez 4 , Francisco Sole 6,7 , Maria A. Garcia 8 , Macarena Peran 4,10 , Rafael Rosell 9 , Juan A. Marchal 10,11 and Jose A. Lorente 1,2 1 GENYO. Pfizer-University of Granada-Andalusian Government Centre of Genomics and Oncology, Granada, Spain 2 Laboratory of Genetic Identification-UGR, Department of Legal Medicine, University of Granada, Granada, Spain 3 Hospital de Barcelona, Medical Oncology Department, Barcelona, Spain 4 University of Jaen, Division of Preventive Medicine and Public Health, CIBERESP, Jaen, Spain 5 Molecular Cytogenetics Laboratory; Pathology Department, Parc de Salut Mar-Hospital del Mar-IMIM-GRETNHE, Barcelona, Spain 6 Medicine Department. Universitat Autonoma de Barcelona, Barcelona, Spain 7 Institut de Recerca contra la Leucemia Josep Carreras, Badalona, Spain 8 Department of Oncology, Virgen de las Nieves, University Hospital, Granada, Spain 9 Catalan Institute of Oncology, Hospital Germans Trias i Pujol, Badalona, Spain (RR); Pangaea Biotech SL, USP Dexeus University Institute, Barcelona, Spain (RR, MAM) 10 Biopathology and Regenerative Medicine Institute (IBIMER), Centre for Biomedical Research, University of Granada, Granada, Spain 11 Department of Human Anatomy and Embryology, University of Granada, Granada, Spain * These authors contributed equally to this work Correspondence: Jose A Lorente, email: // Juan Antonio Marchal, email: // Keywords : Breast Cancer, Circulating Tumor Cells, EGFR, Epithelial-Mesenchymal Transition, Vimentin, Slug, Bcl-2, Apoptosis Received : May 16, 2014 Accepted : July 13, 2014 Published : July 14, 2014 Abstract Circulating tumor cells (CTCs) are frequently associated with epithelial-mesenchymal transition (EMT).The objective of this study was to detect EMT phenotype through Vimentin (VIM) and Slug expression in cytokeratin (CK)-negative CTCs in non-metastatic breast cancer patients and to determine the importance of EGFR in the EMT phenomenon. In CK-negative CTCs samples, both VIM and Slug markers were co-expressed in the most of patients. Among patients EGFR+, half of them were positive for these EMT markers. Furthermore, after a systemic treatment 68% of patients switched from CK- to CK+ CTCs. In our experimental model we found that activation of EGFR signaling by its ligand on MCF-7 cells is sufficient to increase EMT phenotypes, to inhibit apoptotic events and to induce the loss of CK expression. The simultaneous detection of both EGFR and EMT markers in CTCs may improve prognostic or predictive information in patients with operable breast cancer.

32. HER2-signaling pathway, JNK and ERKs kinases, and cancer stem-like cells are targets of Bozepinib small compound

Author

Ramirez, Alberto, Boulaiz, Houria, Morata-Tarifa, Cynthia, Peran, Macarena, Jimenez, Gema, Picon-Ruiz, Manuel, Agil, Ahmad, Cruz-Lopez, Olga, Conejo-Garcia, Ana, Joaquin Campos, Sanchez, Ana, Garcia, Maria A., Marchal, Juan A., [Ramirez,A, Perán,M] Department of Health Sciences, University of Jaén, Jaén, Spain. [Boulaiz,H, Jiménez,G, Marchal,JA] Department of Human Anatomy and Embryology, University of Granada, Granada, Spain. [Boulaiz,H, Morata-Tarifa,C, Jimenez,G, Picon-Ruiz,M, García,MA, Marchal,JA] Biopathology and Medicine Regenerative Institute (IBIMER), University of Granada, Granada, Spain. [Boulaiz.H, Cruz-López,O, Conejo-García,A, Campos,JM, Marchal,JA] Biosanitary Institute of Granada ,Hospitales Universitarios de Granada-Universidad de Granada, Granada, Spain. [Picon-Ruiz,M] Braman Family Breast Cancer Institute, Sylvester Comprehensive Cancer Center, University of Miami, Miller School of Medicine, Miami, FL, USA. [Agil,A] Department of Pharmacology and Neurosciences Institute, Faculty of Medicine, Spain. [Cruz-López, Campos,JM] Department of Pharmaceutical and Organic Chemistry, Faculty of Pharmacy, University of Granada, Granada, Spain. [Sánchez,A] Andalusian Public Health System Biobank, Granada, Spain. [García,MA] Department of Oncology, Virgen de las Nieves, University Hospital, Granada, Spain., and This work was supported in part by grants from the Instituto de Salud Carlos III (Fondo de Investigación Sanitaria, projects nº. PI10/02295, CP08/00063 and PI10/00592) and the ERDF (European Regional Development Fund).

Subjects
Carcinogénesis, Regulación hacia abajo, Piranos, beta Catenina, Anatomy::Cells::Stem Cells::Neoplastic Stem Cells [Medical Subject Headings], Bozepinib, Chemicals and Drugs::Heterocyclic Compounds::Heterocyclic Compounds, 2-Ring::Purines [Medical Subject Headings], Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Insectivora::Hedgehogs [Medical Subject Headings], Chemicals and Drugs::Heterocyclic Compounds::Pyrans [Medical Subject Headings], Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Peptides::Intercellular Signaling Peptides and Proteins::Angiogenic Proteins::Vascular Endothelial Growth Factors::Vascular Endothelial Growth Factor A [Medical Subject Headings], Metastasis, Ratones desnudos, Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings], Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Transferases::Phosphotransferases::Phosphotransferases (Alcohol Group Acceptor)::Protein Kinases::Protein-Tyrosine Kinases::Receptor Protein-Tyrosine Kinases::Receptor, erbB-2 [Medical Subject Headings], Protein kinases, Receptor ErbB-2, Regulación hacia arriba, Organisms::Eukaryota::Animals [Medical Subject Headings], Neoplasias del colon, Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Armadillo Domain Proteins::beta Catenin [Medical Subject Headings], Recurrencia local de neoplasia, Células madre neoplásicas, Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Rodentia::Muridae::Murinae::Mice [Medical Subject Headings], Phenomena and Processes::Chemical Phenomena::Biochemical Phenomena::Biochemical Processes::Down-Regulation [Medical Subject Headings], Phenomena and Processes::Chemical Phenomena::Biochemical Phenomena::Biochemical Processes::Up-Regulation [Medical Subject Headings], Chemicals and Drugs::Heterocyclic Compounds::Heterocyclic Compounds, 1-Ring::Azepines::Oxazepines [Medical Subject Headings], Erizos, Femenino, Diseases::Neoplasms::Neoplastic Processes::Neoplasm Recurrence, Local [Medical Subject Headings], Phenomena and Processes::Chemical Phenomena::Biochemical Phenomena::Biochemical Processes::Signal Transduction [Medical Subject Headings], Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Rodentia::Muridae::Murinae::Mice::Mice, Mutant Strains::Mice, Nude [Medical Subject Headings], Cancer stem-like cells, Diseases::Neoplasms::Neoplastic Processes::Cocarcinogenesis [Medical Subject Headings], Humanos, Factor A de crecimiento endotelial vascular, Check Tags::Female [Medical Subject Headings], HER-2, Oxazepinas, Transducción de señal, Animales, Diseases::Neoplasms::Neoplasms by Site::Digestive System Neoplasms::Gastrointestinal Neoplasms::Intestinal Neoplasms::Colorectal Neoplasms::Colonic Neoplasms [Medical Subject Headings], Purinas
Abstract

Journal Article; Research Support, Non-U.S. Gov't; Identification of novel anticancer drugs presenting more than one molecular target and efficacy against cancer stem-like cells (CSCs) subpopulations represents a therapeutic need to combat the resistance and the high risk of relapse in patients. In the present work we show how Bozepinib [(RS)-2,6-dichloro-9-[1-(p-nitrobenzenesulfonyl)-1,2,3,5-tetrahydro-4,1-benzoxazepin-3-yl]-9H-purine], a small anti-tumor compound, demonstrated selectivity on cancer cells and showed an inhibitory effect over kinases involved in carcinogenesis, proliferation and angiogenesis. The cytotoxic effects of Bozepinib were observed in both breast and colon cancer cells expressing different receptor patterns. Bozepinib inhibited HER-2 signaling pathway and JNK and ERKs kinases. In addition, Bozepinib has an inhibitory effect on AKT and VEGF together with anti-angiogenic and anti-migratory activities. Moreover, the modulation of pathways involved in tumorigenesis by Bozepinib was also evident in microarrays analysis. Interestingly, Bozepinib inhibited both mamo- and colono-spheres formation and eliminated ALDH+ CSCs subpopulations at a low micromolar range similar to Salinomycin. Bozepinib induced the down-regulation of c-MYC, β-CATENIN and SOX2 proteins and the up-regulation of the GLI-3 hedgehog-signaling repressor. Finally, Bozepinib shows in vivo anti-tumor and anti-metastatic efficacy in xenotransplanted nude mice without presenting sub-acute toxicity. These findings support further studies on the therapeutic potential of Bozepinib in cancer patients. Yes

33. The Chemotherapeutic Drug 5-Fluorouracil Promotes PKR-Mediated Apoptosis in a p53- Independent Manner in Colon and Breast Cancer Cells

Author

Mariano Esteban, María Ángel García, Antonia Aránega, Margarita Aguilera, Joaquín M. Campos, Esther Carrasco, Jose Prados, Carmen Rivas, Miguel Angel Calleja, Pablo J. Álvarez, Juan A. Marchal, [García,MA] Unidad de Investigación, Hospital Universitario Virgen de las Nieves, Granada, Spain.[Aguilera,M] Unidad de Farmacogenética, Hospital Universitario Virgen de las Nieves, Granada, Spain.[García,MA, Carrasco,E, Alvarez,P, Prados,JC, Marchal,JA, Aránega,A] Instituto de Biopatología y Medicina Regenerativa (IBIMER), Universidad de Granada,Granada, Spain. [Rivas,C] Centro Nacional de Biotecnología, CSIC, Madrid,Spain. [Campos,JM] Departamento de Química Farmacéutica y Orgánica, Facultad de Farmacia, Universidad de Granada, Granada, Spain., and This work was supported in part by grants from the Instituto de Salud Carlos III (Fondo de Investigación Sanitaria projects n°. CP08/0063, PI07/0527 and PI10/02295). No additional external funding was received for this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Subjects
Anatomy::Cells::Cells, Cultured::Cell Line::Cell Line, Tumor [Medical Subject Headings], Colorectal cancer, Cancer Treatment, lcsh:Medicine, Apoptosis, Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings], eIF-2 Kinase, Breast cancer, Molecular Cell Biology, Cytotoxic T cell, Chemicals and Drugs::Heterocyclic Compounds::Heterocyclic Compounds, 1-Ring::Pyrimidines::Pyrimidinones::Uracil::Fluorouracil [Medical Subject Headings], Flow cytometry, Phosphorylation, lcsh:Science, Phenomena and Processes::Metabolic Phenomena::Metabolism::Phosphorylation [Medical Subject Headings], Cellular Stress Responses, Multidisciplinary, biology, medicine.diagnostic_test, Cell Death, Chemistry, Colon Adenocarcinoma, Neoplasias de la Mama, Femenino, Obstetrics and Gynecology, Humanos, Antimetabolitos Antineoplásicos, Oncology, Cancer treatment, Colonic Neoplasms, Medicine, Female, Fluorouracil, Fosforilación, Research Article, Programmed cell death, Antimetabolites, Antineoplastic, Drugs and Devices, Colon, Breast Neoplasms, Gastroenterology and Hepatology, eIF-2 Quinasa, Cell Line, Tumor, Breast Cancer, Gastrointestinal Tumors, medicine, Humans, Protein kinase A, Biology, Diseases::Neoplasms::Neoplasms by Site::Breast Neoplasms [Medical Subject Headings], EIF-2 kinase, lcsh:R, Neoplasias del Colon, Cancers and Neoplasms, Chemotherapy and Drug Treatment, medicine.disease, Protein kinase R, Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Transferases::Phosphotransferases::Phosphotransferases (Alcohol Group Acceptor)::Protein Kinases::Protein-Serine-Threonine Kinases::eIF-2 Kinase [Medical Subject Headings], Chemicals and Drugs::Chemical Actions and Uses::Pharmacologic Actions::Molecular Mechanisms of Pharmacological Action::Antimetabolites::Antimetabolites, Antineoplastic [Medical Subject Headings], SW480 cells, Línea Celular Tumoral, Immunology, Diseases::Neoplasms::Neoplasms by Site::Digestive System Neoplasms::Gastrointestinal Neoplasms::Intestinal Neoplasms::Colorectal Neoplasms::Colonic Neoplasms [Medical Subject Headings], Cancer research, biology.protein, lcsh:Q, Tumor Suppressor Protein p53
Abstract

The chemotherapeutic drug 5-FU is widely used in the treatment of a range of cancers, but resistance to the drug remains a major clinical problem. Since defects in the mediators of apoptosis may account for chemo-resistance, the identification of new targets involved in 5-FU-induced apoptosis is of main clinical interest. We have identified the ds-RNA-dependent protein kinase (PKR) as a key molecular target of 5-FU involved in apoptosis induction in human colon and breast cancer cell lines. PKR distribution and activation, apoptosis induction and cytotoxic effects were analyzed during 5-FU and 5-FU/IFNα treatment in several colon and breast cancer cell lines with different p53 status. PKR protein was activated by 5-FU treatment in a p53-independent manner, inducing phosphorylation of the protein synthesis translation initiation factor eIF-2α and cell death by apoptosis. Furthermore, PKR interference promoted a decreased response to 5-FU treatment and those cells were not affected by the synergistic antitumor activity of 5-FU/IFNα combination. These results, taken together, provide evidence that PKR is a key molecular target of 5-FU with potential relevance in the clinical use of this drug., This work was supported in part by grants from the Instituto de Salud Carlos III (Fondo de Investigación Sanitaria projects n°. CP08/0063, PI07/0527 and PI10/02295).

Published
2011

34. Sex determination and gonadal development in the common toad (Bufo bufo)

Author

Roco, Alvaro, La Guardia Quiles, Rafael Diaz, Falconi, Rosanna, Zacanti, Francesco, David Durusel, Jean, Antonio Marchal, Juan, Antonio Sánchez Baca, Bullejos, Monica, JALLON J-M, LAFONT R., D'HONDT J.L., Roco A., Diaz de la Guardia Quiles R., Falconi R., Zaccanti F., Durussel J.D., Marchal J.A., Sanchez A., Bullejos M., Roco, A, Quiles, Rdd, Falconi, Rosanna, Zaccanti, Francesco, Durussel, JEAN-DAVID, Marchal, Ja, Sanchez, A, and Bullejos, M.

Subjects
SEX DETERMINATION, endocrine system, GONADAL DIFFERENTIATION, urogenital system, BUFO BUFO, AMPHIBIA, sense organs, Bufo bufo, Sex Determination, Gonadal Development, SOX9, SOX9
Abstract

Amphibians have been traditionally used as animal models in embryology. Nevertheless, the knowledge about the mechanisms involved in sex determination and differentiation are very scarce in this group. In the species Bufo bufo there are no sex chromosomes morphologically distinguishable. However, by analysing the sex ratio of the offspring of sex reversed animals it is possible to conclude that the females are the heterogametic sex in this species. That is, the species Bufo bufo has a ZZ/ZW chromosomes system for sex determination. Regarding to gonadal development, the species of the genus Bufo, have the particularity of developing undifferentiated ovaries (Bidder's organs) in the cranial portion of male and female gonads. To study gonadal development in B. Bufo, we are cloning in this species orthologous genes to those involved in gonadal development in other vertebrate groups. The study of the expression pattern of these genes during gonadal differentiation, and its relation with the morphological changes that take place, will enable us to establish their role in this process. SOX9 gene, involved in Sertoli cell differentiation, is among the genes we are studying. This gene codes for a transcription factor closely related to SRY gene, is male-specific and necessary for testis differentiation. In B. bufo this gene is expressed only in developing male gonads, starting at the stage where morphological differences start to be observed between developing testes and ovaries.

Published
2008

35. Using deep learning for predicting the dynamic evolution of breast cancer migration.

Author

Garcia-Moreno FM, Ruiz-Espigares J, Gutiérrez-Naranjo MA, and Marchal JA

Subjects
Humans, Female, MCF-7 Cells, Models, Biological, Wound Healing, Deep Learning, Breast Neoplasms pathology, Cell Movement
Abstract

Background: Breast cancer (BC) remains a prevalent health concern, with metastasis as the main driver of mortality. A detailed understanding of metastatic processes, particularly cell migration, is fundamental to improve therapeutic strategies. The wound healing assay, a traditional two-dimensional (2D) model, offers insights into cell migration but presents scalability issues due to data scarcity, arising from its manual and labor-intensive nature., Method: To overcome these limitations, this study introduces the Prediction Wound Progression Framework (PWPF), an innovative approach utilizing Deep Learning (DL) and artificial data generation. The PWPF comprises a DL model initially trained on artificial data that simulates wound healing in MCF-7 BC cell monolayers and spheres, which is subsequently fine-tuned on real-world data., Results: Our results underscore the model's effectiveness in analyzing and predicting cell migration dynamics within the wound healing context, thus enhancing the usability of 2D models. The PWPF significantly contributes to a better understanding of cell migration processes in BC and expands the possibilities for research into wound healing mechanisms., Conclusions: These advancements in automated cell migration analysis hold the potential for more comprehensive and scalable studies in the future. Our dataset, models, and code are publicly available at https://github.com/frangam/wound-healing., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Juan Antonio Marchal reports financial support was provided by University of Granada. Miguel Angel Guterrez-Naranjo reports financial support was provided by University of Seville. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Published
2024
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36. Onychocolone A produced by the fungus Onychocola sp. targets cancer stem cells and stops pancreatic cancer progression by inhibiting MEK2-dependent cell signaling.

Author

Ramos MC, Crespo-Sueiro G, de Pedro N, Griñán-Lisón C, Díaz C, Pérez-Victoria I, González-Menéndez V, Castillo F, Pérez Del Palacio J, Tormo JR, Choquesillo-Lazarte D, Marchal JA, Vicente F, Fernández-Godino R, Genilloud O, and Reyes F

Subjects
Animals, Humans, Cell Line, Tumor, Mice, Benzophenones pharmacology, Benzophenones chemistry, Xenograft Model Antitumor Assays, Ascomycota chemistry, Mice, Nude, Cell Proliferation drug effects, Pancreatic Neoplasms drug therapy, Pancreatic Neoplasms pathology, Neoplastic Stem Cells drug effects, Neoplastic Stem Cells pathology, Neoplastic Stem Cells metabolism, Signal Transduction drug effects, Antineoplastic Agents pharmacology
Abstract

Pancreatic cancer (PC) shows a high fatality rate that can only be faced with a combination of surgery and chemotherapy or palliative treatment in the case of advanced patients. Besides, PC tumors are enriched with subpopulations of cancer stem cells (CSCs) that are resistant to the existing chemotherapeutic agents, which raises an important need for the identification of new drugs. To fill this gap, we have tested the anti-tumoral activity of microbial extracts, which chemical diversity offers a broad spectrum of potential new bioactive compounds. Extracts derived from the fungus Onychocola sp. CF-107644 were assayed via high throughput screening followed by bioassay-guided fractionation and resulted in the identification and isolation of six benzophenone derivatives with antitumoral activity: onychocolones A-F (#1-6). The structures of the compounds were established by spectroscopic methods, including ESI-TOF MS, 1D and 2D NMR analyses and X-ray diffraction. Compounds #1-4 significantly inhibited the growth of the pancreas tumoral cell lines, with low-micromolar Median Effective Doses (ED 50 s). Compound #1 (onychocolone A) was prioritized for further profiling due to its pro-apoptotic effect, which was further validated on 3D spheroids and pancreatic CSCs. Protein expression assays showed that the effect was mechanistically linked to the inhibition of MEK onco-signaling pathway. The efficacy of onychocolone A was also demonstrated in vivo by the reduction of tumor growth in a pancreatic xenograft mouse model generated by CSCs. Altogether, the data support that onychocolone A is a promising new small molecule for hit-to-lead development of a new treatment for PC., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Published
2024
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37. Design and evaluation of a bilayered dermal/hypodermal 3D model using a biomimetic hydrogel formulation.

Author

Chocarro-Wrona C, López de Andrés J, Rioboó-Legaspi P, Pleguezuelos-Beltrán P, Antich C, De Vicente J, Gálvez-Martín P, López-Ruiz E, and Marchal JA

Subjects
Humans, Cell Survival drug effects, Mesenchymal Stem Cells drug effects, Hyaluronic Acid chemistry, Hyaluronic Acid pharmacology, Wound Healing drug effects, Collagen Type I metabolism, Skin drug effects, Skin metabolism, Dermatan Sulfate chemistry, Dermatan Sulfate pharmacology, Fibroblasts drug effects, Elastin chemistry, Extracellular Matrix metabolism, Biomimetics methods, Sepharose chemistry, Dermis drug effects, Dermis metabolism, Dermis cytology, Animals, Hydrogels chemistry, Biomimetic Materials chemistry, Biomimetic Materials pharmacology, Tissue Engineering methods
Abstract

Due to the limitations of the current skin wound treatments, it is highly valuable to have a wound healing formulation that mimics the extracellular matrix (ECM) and mechanical properties of natural skin tissue. Here, a novel biomimetic hydrogel formulation has been developed based on a mixture of Agarose-Collagen Type I (AC) combined with skin ECM-related components: Dermatan sulfate (DS), Hyaluronic acid (HA), and Elastin (EL) for its application in skin tissue engineering (TE). Different formulations were designed by combining AC hydrogels with DS, HA, and EL. Cell viability, hemocompatibility, physicochemical, mechanical, and wound healing properties were investigated. Finally, a bilayered hydrogel loaded with fibroblasts and mesenchymal stromal cells was developed using the Ag-Col I-DS-HA-EL (ACDHE) formulation. The ACDHE hydrogel displayed the best in vitro results and acceptable physicochemical properties. Also, it behaved mechanically close to human native skin and exhibited good cytocompatibility. Environmental scanning electron microscopy (ESEM) analysis revealed a porous microstructure that allows the maintenance of cell growth and ECM-like structure production. These findings demonstrate the potential of the ACDHE hydrogel formulation for applications such as an injectable hydrogel or a bioink to create cell-laden structures for skin TE., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Patricia Galvez-Martin reports a relationship with Bioibérica SA that includes: employment. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2024 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Published
2024
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38. Beyond stiffness: deciphering the role of viscoelasticity in cancer evolution and treatment response.

Author

Zubiarrain-Laserna A, Martínez-Moreno D, López de Andrés J, de Lara-Peña L, Guaresti O, Zaldua AM, Jiménez G, and Marchal JA

Subjects
Humans, Viscosity, Animals, Hydrogels chemistry, Biomechanical Phenomena, Neoplasms pathology, Neoplasms drug therapy, Elasticity, Extracellular Matrix metabolism
Abstract

There is increasing evidence that cancer progression is linked to tissue viscoelasticity, which challenges the commonly accepted notion that stiffness is the main mechanical hallmark of cancer. However, this new insight has not reached widespread clinical use, as most clinical trials focus on the application of tissue elasticity and stiffness in diagnostic, therapeutic, and surgical planning. Therefore, there is a need to advance the fundamental understanding of the effect of viscoelasticity on cancer progression, to develop novel mechanical biomarkers of clinical significance. Tissue viscoelasticity is largely determined by the extracellular matrix (ECM), which can be simulated in vitro using hydrogel-based platforms. Since the mechanical properties of hydrogels can be easily adjusted by changing parameters such as molecular weight and crosslinking type, they provide a platform to systematically study the relationship between ECM viscoelasticity and cancer progression. This review begins with an overview of cancer viscoelasticity, describing how tumor cells interact with biophysical signals in their environment, how they contribute to tumor viscoelasticity, and how this translates into cancer progression. Next, an overview of clinical trials focused on measuring biomechanical properties of tumors is presented, highlighting the biomechanical properties utilized for cancer diagnosis and monitoring. Finally, this review examines the use of biofabricated tumor models for studying the impact of ECM viscoelasticity on cancer behavior and progression and it explores potential avenues for future research on the production of more sophisticated and biomimetic tumor models, as well as their mechanical evaluation., (Creative Commons Attribution license.)

Published
2024
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39. Deciphering the performance of macrophages in tumour microenvironment: a call for precision immunotherapy.

Author

Toledo B, Zhu Chen L, Paniagua-Sancho M, Marchal JA, Perán M, and Giovannetti E

Subjects
Humans, Precision Medicine, Macrophages immunology, Animals, Tumor Microenvironment immunology, Immunotherapy methods, Neoplasms immunology, Neoplasms therapy, Neoplasms pathology, Tumor-Associated Macrophages immunology
Abstract

Macrophages infiltrating tumour tissues or residing in the microenvironment of solid tumours are known as tumour-associated macrophages (TAMs). These specialized immune cells play crucial roles in tumour growth, angiogenesis, immune regulation, metastasis, and chemoresistance. TAMs encompass various subpopulations, primarily classified into M1 and M2 subtypes based on their differentiation and activities. M1 macrophages, characterized by a pro-inflammatory phenotype, exert anti-tumoural effects, while M2 macrophages, with an anti-inflammatory phenotype, function as protumoural regulators. These highly versatile cells respond to stimuli from tumour cells and other constituents within the tumour microenvironment (TME), such as growth factors, cytokines, chemokines, and enzymes. These stimuli induce their polarization towards one phenotype or another, leading to complex interactions with TME components and influencing both pro-tumour and anti-tumour processes.This review comprehensively and deeply covers the literature on macrophages, their origin and function as well as the intricate interplay between macrophages and the TME, influencing the dual nature of TAMs in promoting both pro- and anti-tumour processes. Moreover, the review delves into the primary pathways implicated in macrophage polarization, examining the diverse stimuli that regulate this process. These stimuli play a crucial role in shaping the phenotype and functions of macrophages. In addition, the advantages and limitations of current macrophage based clinical interventions are reviewed, including enhancing TAM phagocytosis, inducing TAM exhaustion, inhibiting TAM recruitment, and polarizing TAMs towards an M1-like phenotype. In conclusion, while the treatment strategies targeting macrophages in precision medicine show promise, overcoming several obstacles is still necessary to achieve an accessible and efficient immunotherapy., (© 2024. The Author(s).)

Published
2024
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40. Can Hemorrhagic Stroke Genetics Help Forensic Diagnosis in Pediatric Age (<5 Years Old)?

Author

Treves B, Sonnini E, La Russa R, Del Duca F, Ghamlouch A, De Matteis A, Trignano C, Marchal JA, Carrillo E, Napoletano G, and Maiese A

Subjects
Humans, Child, Preschool, Genetic Testing methods, Craniocerebral Trauma diagnosis, Craniocerebral Trauma genetics, Infant, Diagnosis, Differential, Hemorrhagic Stroke genetics, Hemorrhagic Stroke diagnosis
Abstract

When stroke occurs in pediatric age, it might be mistakenly interpreted as non-accidental head injury (NAHI). In these situations, a multidisciplinary approach is fundamental, including a thorough personal and familial history, along with accurate physical examination and additional investigations. Especially when the clinical picture is uncertain, it is important to remember that certain genetic conditions can cause bleeding inside the brain, which may resemble NAHI. Pediatric strokes occurring around the time of birth can also be an initial sign of undiagnosed genetic disorders. Hence, it is crucial to conduct a thorough evaluation, including genetic testing, when there is a suspicion of NAHI but the symptoms are unclear. In these cases, a characteristic set of symptoms is often observed. This study aims to summarize some of the genetic causes of hemorrhagic stroke in the pediatric population, thus mimicking non-accidental head injury, considering elements that can be useful in characterizing pathologies. A systematic review of genetic disorders that may cause ICH in children was carried out according to the Preferred Reporting Item for Systematic Review (PRISMA) standards. We selected 10 articles regarding the main genetic diseases in stroke; we additionally selected 11 papers concerning patients with pediatric stroke and genetic diseases, or studies outlining the characteristics of stroke in these patients. The disorders we identified were Moyamoya disease (MMD), COL4A1 , COL4A2 pathogenic variant, Ehlers-Danlos syndrome (E-D), neurofibromatosis type 1 (Nf1), sickle cell disease (SCD), cerebral cavernous malformations (CCM), hereditary hemorrhagic telangiectasia (HHT) and Marfan syndrome. In conclusion, this paper provides a comprehensive overview of the genetic disorders that could be tested in children when there is a suspicion of NAHI but an unclear picture.

Published
2024
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41. Autologous patient-derived exhausted nano T-cells exploit tumor immune evasion to engage an effective cancer therapy.

Author

Blaya-Cánovas JL, Griñán-Lisón C, Blancas I, Marchal JA, Ramírez-Tortosa C, López-Tejada A, Benabdellah K, Cortijo-Gutiérrez M, Cano-Cortés MV, Graván P, Navarro-Marchal SA, Gómez-Morales J, Delgado-Almenta V, Calahorra J, Agudo-Lera M, Sagarzazu A, Rodríguez-González CJ, Gallart-Aragón T, Eich C, Sánchez-Martín RM, and Granados-Principal S

Subjects
Humans, Animals, Mice, Female, Cell Line, Tumor, Immune Evasion, Triple Negative Breast Neoplasms therapy, Triple Negative Breast Neoplasms immunology, Triple Negative Breast Neoplasms pathology, Xenograft Model Antitumor Assays, Nanoparticles chemistry, T-Lymphocytes immunology, T-Lymphocytes metabolism
Abstract

Background: Active targeting by surface-modified nanoplatforms enables a more precise and elevated accumulation of nanoparticles within the tumor, thereby enhancing drug delivery and efficacy for a successful cancer treatment. However, surface functionalization involves complex procedures that increase costs and timelines, presenting challenges for clinical implementation. Biomimetic nanoparticles (BNPs) have emerged as unique drug delivery platforms that overcome the limitations of actively targeted nanoparticles. Nevertheless, BNPs coated with unmodified cells show reduced functionalities such as specific tumor targeting, decreasing the therapeutic efficacy. Those challenges can be overcome by engineering non-patient-derived cells for BNP coating, but these are complex and cost-effective approaches that hinder their wider clinical application. Here we present an immune-driven strategy to improve nanotherapeutic delivery to tumors. Our unique perspective harnesses T-cell exhaustion and tumor immune evasion to develop a groundbreaking new class of BNPs crafted from exhausted T-cells (NExT) of triple-negative breast cancer (TNBC) patients by specific culture methods without sophisticated engineering., Methods: NExT were generated by coating PLGA (poly(lactic-co-glycolic acid)) nanoparticles with TNBC-derived T-cells exhausted in vitro by acute activation. Physicochemical characterization of NExT was made by dynamic light scattering, electrophoretic light scattering and transmission electron microscopy, and preservation and orientation of immune checkpoint receptors by flow cytometry. The efficacy of chemotherapy-loaded NExT was assessed in TNBC cell lines in vitro. In vivo toxicity was made in CD1 mice. Biodistribution and therapeutic activity of NExT were determined in cell-line- and autologous patient-derived xenografts in immunodeficient mice., Results: We report a cost-effective approach with a good performance that provides NExT naturally endowed with immune checkpoint receptors (PD1, LAG3, TIM3), augmenting specific tumor targeting by engaging cognate ligands, enhancing the therapeutic efficacy of chemotherapy, and disrupting the PD1/PDL1 axis in an immunotherapy-like way. Autologous patient-derived NExT revealed exceptional intratumor accumulation, heightened chemotherapeutic index and efficiency, and targeted the tumor stroma in a PDL1 + patient-derived xenograft model of triple-negative breast cancer., Conclusions: These advantages underline the potential of autologous patient-derived NExT to revolutionize tailored adoptive cancer nanotherapy and chemoimmunotherapy, which endorses their widespread clinical application of autologous patient-derived NExT., (© 2024. The Author(s).)

Published
2024
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42. Unlocking the effective alliance of β-lapachone and hydroxytyrosol against triple-negative breast cancer cells.

Author

Calahorra J, Blaya-Cánovas JL, Castellini-Pérez O, Aparicio-Puerta E, Cives-Losada C, Marin JJG, Rementeria M, Cara FE, López-Tejada A, Griñán-Lisón C, Aulicino F, Berger I, Marchal JA, Delgado-Almenta V, and Granados-Principal S

Subjects
Humans, Cell Line, Tumor, Female, Drug Synergism, Neoplastic Stem Cells drug effects, Neoplastic Stem Cells metabolism, Neoplastic Stem Cells pathology, Drug Resistance, Neoplasm drug effects, Cell Cycle Checkpoints drug effects, Triple Negative Breast Neoplasms drug therapy, Triple Negative Breast Neoplasms pathology, Triple Negative Breast Neoplasms metabolism, Naphthoquinones pharmacology, Phenylethyl Alcohol analogs & derivatives, Phenylethyl Alcohol pharmacology, Apoptosis drug effects, NAD(P)H Dehydrogenase (Quinone) metabolism, NAD(P)H Dehydrogenase (Quinone) genetics, Cell Proliferation drug effects
Abstract

Triple-negative breast cancer (TNBC) is characterised by its aggressiveness and resistance to chemotherapy, demanding the development of effective strategies against its unique characteristics. Derived from lapacho tree bark, β-lapachone (β-LP) selectively targets cancer cells with elevated levels of the detoxifying enzyme NQO1. Hydroxytyrosol (HT) is a phenolic compound derived from olive trees with important anticancer properties that include the inhibition of cancer stem cells (CSCs) and metastatic features in TNBC, as well as relevant antioxidant activities by mechanisms such as the induction of NQO1. We aimed to study whether these compounds could have synergistic anticancer activity in TNBC cells and the possible role of NQO1. For this pourpose, we assessed the impact of β-LP (0.5 or 1.5 μM) and HT (50 and 100 μM) on five TNBC cell lines. We demonstrated that the combination of β-LP and HT exhibits anti-proliferative, pro-apoptotic, and cell cycle arrest effects in several TNBC cells, including docetaxel-resistant TNBC cells. Additionally, it effectively inhibits the self-renewal and clonogenicity of CSCs, modifying their aggressive phenotype. However, the notable impact of the β-LP-HT combination does not appear to be solely associated with the levels of the NQO1 protein and ROS. RNA-Seq analysis revealed that the combination's anticancer activity is linked to a strong induction of endoplasmic reticulum stress and apoptosis through the unfolded protein response. In conclusion, in this study, we demonstrated how the combination of β-LP and HT could offer an affordable, safe, and effective approach against TNBC., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Published
2024
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43. Towards a More Efficient Breast Cancer Therapy Using Active Human Cell Membrane-Coated Metal-Organic Frameworks.

Author

Graván P, Rojas S, Picchi DF, Galisteo-González F, Horcajada P, and Marchal JA

Abstract

The recent description of well-defined molecular subtypes of breast cancer has led to the clinical development of a number of successful molecular targets. Particularly, triple-negative breast cancer (TNBC) is an aggressive type of breast cancer with historically poor outcomes, mainly due to the lack of effective targeted therapies. Recent progresses in materials science have demonstrated the impressive properties of metal-organic framework nanoparticles (NPs) as antitumoral drug delivery systems. Here, in a way to achieve efficient bio-interfaces with cancer cells and improve their internalization, benchmarked MIL-100(Fe) NPs were coated with cell membranes (CMs) derived from the human TNBC cell line MDA-MB-468. The prepared CMs-coated metal-organic framework (CMs&#95;MIL-100(Fe)) showed enhanced colloidal stability, cellular uptake, and cytotoxicity in MDA-MB-468 cells compared to non-coated NPs, paving the way for these human CMs-coated MIL-100(Fe) NPs as effective targeted therapies against the challenging TNBC.

Published
2024
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44. Human mesenchymal stromal cells-laden crosslinked hyaluronic acid-alginate bioink for 3D bioprinting applications in tissue engineering.

Author

Galocha-León C, Antich C, Voltes-Martínez A, Marchal JA, Mallandrich M, Halbaut L, Souto EB, Gálvez-Martín P, and Clares-Naveros B

Abstract

Three-dimensional (3D) bioprinting is considered one of the most advanced tools to build up materials for tissue engineering. The aim of this work was the design, development and characterization of a bioink composed of human mesenchymal stromal cells (hMSC) for extrusion through nozzles to create these 3D structures that might potentially be apply to replace the function of damaged natural tissue. In this study, we focused on the advantages and the wide potential of biocompatible biomaterials, such as hyaluronic acid and alginate for the inclusion of hMSC. The bioink was characterized for its physical (pH, osmolality, degradation, swelling, porosity, surface electrical properties, conductivity, and surface structure), mechanical (rheology and printability) and biological (viability and proliferation) properties. The developed bioink showed high porosity and high swelling capacity, while the degradation rate was dependent on the temperature. The bioink also showed negative electrical surface and appropriate rheological properties required for bioprinting. Moreover, stress-stability studies did not show any sign of physical instability. The developed bioink provided an excellent environment for the promotion of the viability and growth of hMSC cells. Our work reports the first-time study of the effect of storage temperature on the cell viability of bioinks, besides showing that our bioink promoted a high cell viability after being extruded by the bioprinter. These results support the suggestion that the developed hMSC-composed bioink fulfills all the requirements for tissue engineering and can be proposed as a biological tool with potential applications in regenerative medicine and tissue engineering., (© 2024. The Author(s).)

Published
2024
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45. Identification of novel biomarkers in the early diagnosis of malignant melanoma by untargeted liquid chromatography coupled to high-resolution mass spectrometry-based metabolomics: a pilot study.

Author

Peña-Martín J, Belén García-Ortega M, Palacios-Ferrer JL, Díaz C, Ángel García M, Boulaiz H, Valdivia J, Jurado JM, Almazan-Fernandez FM, Arias Santiago S, Vicente F, Del Val C, Pérez Del Palacio J, and Marchal JA

Subjects
Humans, Pilot Projects, Early Detection of Cancer, Metabolomics, Biomarkers, Liquid Chromatography-Mass Spectrometry, Melanoma, Skin Neoplasms
Abstract

Background: Malignant melanoma (MM) is a highly aggressive form of skin cancer whose incidence continues to rise worldwide. If diagnosed at an early stage, it has an excellent prognosis, but mortality increases significantly at advanced stages after distant spread. Unfortunately, early detection of aggressive melanoma remains a challenge., Objectives: To identify novel blood-circulating biomarkers that may be useful in the diagnosis of MM to guide patient counselling and appropriate disease management., Methods: In this study, 105 serum samples from 26 healthy patients and 79 with MM were analysed using an untargeted approach by liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS) to compare the metabolomic profiles of both conditions. Resulting data were subjected to both univariate and multivariate statistical analysis to select robust biomarkers. The classification model obtained from this analysis was further validated with an independent cohort of 12 patients with stage I MM., Results: We successfully identified several lipidic metabolites differentially expressed in patients with stage I MM vs. healthy controls. Three of these metabolites were used to develop a classification model, which exhibited exceptional precision (0.92) and accuracy (0.94) when validated on an independent sample., Conclusions: These results demonstrate that metabolomics using LC-HRMS is a powerful tool to identify and quantify metabolites in bodily fluids that could serve as potential early diagnostic markers for MM., Competing Interests: Conflicts of interest The authors declare no conflicts of interest., (© The Author(s) 2024. Published by Oxford University Press on behalf of British Association of Dermatologists.)

Published
2024
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46. Treatment resistance in pancreatic and biliary tract cancer: molecular and clinical pharmacology perspectives.

Author

Toledo B, Deiana C, Scianò F, Brandi G, Marchal JA, Perán M, and Giovannetti E

Subjects
Humans, Artificial Intelligence, Immunotherapy, Combined Modality Therapy, Molecular Targeted Therapy, Tumor Microenvironment, Pharmacology, Clinical, Biliary Tract Neoplasms drug therapy, Biliary Tract Neoplasms genetics, Biliary Tract Neoplasms pathology, Pancreatic Neoplasms drug therapy, Pancreatic Neoplasms genetics
Abstract

Introduction: Treatment resistance poses a significant obstacle in oncology, especially in biliary tract cancer (BTC) and pancreatic cancer (PC). Current therapeutic options include chemotherapy, targeted therapy, and immunotherapy. Resistance to these treatments may arise due to diverse molecular mechanisms, such as genetic and epigenetic modifications, altered drug metabolism and efflux, and changes in the tumor microenvironment. Identifying and overcoming these mechanisms is a major focus of research: strategies being explored include combination therapies, modulation of the tumor microenvironment, and personalized approaches., Areas Covered: We provide a current overview and discussion of the most relevant mechanisms of resistance to chemotherapy, target therapy, and immunotherapy in both BTC and PC. Furthermore, we compare the different strategies that are being implemented to overcome these obstacles., Expert Opinion: So far there is no unified theory on drug resistance and progress is limited. To overcome this issue, individualized patient approaches, possibly through liquid biopsies or single-cell transcriptome studies, are suggested, along with the potential use of artificial intelligence, to guide effective treatment strategies. Furthermore, we provide insights into what we consider the most promising areas of research, and we speculate on the future of managing treatment resistance to improve patient outcomes.

Published
2024
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47. Exploring the Impact of Nanoparticle Stealth Coatings in Cancer Models: From PEGylation to Cell Membrane-Coating Nanotechnology.

Author

Graván P, Peña-Martín J, de Andrés JL, Pedrosa M, Villegas-Montoya M, Galisteo-González F, Marchal JA, and Sánchez-Moreno P

Subjects
Humans, Cell Membrane metabolism, Polyethylene Glycols metabolism, Serum Albumin, Bovine metabolism, Nanomedicine, Nanoparticles metabolism, Neoplasms metabolism
Abstract

Nanotechnological platforms offer advantages over conventional therapeutic and diagnostic modalities. However, the efficient biointerfacing of nanomaterials for biomedical applications remains challenging. In recent years, nanoparticles (NPs) with different coatings have been developed to reduce nonspecific interactions, prolong circulation time, and improve therapeutic outcomes. This study aims to compare various NP coatings to enhance surface engineering for more effective nanomedicines. We prepared and characterized polystyrene NPs with different coatings of poly(ethylene glycol), bovine serum albumin, chitosan, and cell membranes from a human breast cancer cell line. The coating was found to affect the colloidal stability, adhesion, and elastic modulus of NPs. Protein corona formation and cellular uptake of NPs were also investigated, and a 3D tumor model was employed to provide a more realistic representation of the tumor microenvironment. The prepared NPs were found to reduce protein adsorption, and cell-membrane-coated NPs showed significantly higher cellular uptake. The secretion of proinflammatory cytokines in human monocytes after incubation with the prepared NPs was evaluated. Overall, the study demonstrates the importance of coatings in affecting the behavior and interaction of nanosystems with biological entities. The findings provide insight into bionano interactions and are important for the effective implementation of stealth surface engineering designs.

Published
2024
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48. Mimicking the Tumor Niche: Methods for Isolation, Culture, and Characterization of Cancer Stem Cells and Multicellular Spheroids.

Author

De Lara-Peña L, Farace C, Pisano A, de Andrés JL, Fenu G, Etzi F, Griñán-Lisón C, Marchal JA, and Madeddu R

Subjects
Humans, Neoplastic Stem Cells pathology, Tumor Microenvironment, Spheroids, Cellular pathology, Neoplasms pathology
Abstract

Cancer stem cells (CSCs) play a significant role in driving several tumor hallmarks. Their behavior and tumor progression are strictly related to the tumor microenvironment (TME). The dynamic interplay between CSCs and TME drives metastasis, chemoresistance, and disease relapse. In this chapter, we describe different techniques and protocols for isolating, culturing, and characterizing CSCs and we explain the methodology for the culture of multicellular spheroids comprising CSCs., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Published
2024
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49. Metabolomics signature as a survival predictor in patients with resectable colorectal liver metastasis.

Author

González-Olmedo C, García-Verdejo FJ, Reguera-Teba A, Rosa-Garrido C, López-López JA, Díaz-Beltrán L, García PM, Luque-Caro N, Gálvez-Montosa F, Ortega-Granados AL, Ruiz-Sanjuan M, Cózar-Ibáñez A, Sainz J, Marchal JA, Camacho J, Del Palacio JP, Fernández-Godino R, Díaz C, and Sánchez-Rovira P

Subjects
Humans, Oxaliplatin, Colorectal Neoplasms surgery, Colorectal Neoplasms pathology, Liver Neoplasms surgery, Liver Neoplasms secondary
Published
2024
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50. Effect of the Protein Corona Formation on Antibody Functionalized Liquid Lipid Nanocarriers.

Author

Navarro-Marchal SA, Martín-Contreras M, Castro-Santiago D, Del Castillo-Santaella T, Graván P, Jódar-Reyes AB, Marchal JA, and Peula-García JM

Subjects
Humans, Receptor, ErbB-2 metabolism, Antibodies, MCF-7 Cells, Lipids, Protein Corona chemistry, Nanoparticles chemistry
Abstract

The main aim of this study is to report basic knowledge on how a protein corona (PC) could affect or modify the way in which multifunctionalized nanoparticles interact with cells. With this purpose, we have firstly optimized the development of a target-specific nanocarrier by coupling a specific fluorescent antibody on the surface of functionalized lipid liquid nanocapsules (LLNCs). Thus, an anti-HER2-FITC antibody (αHER2) has been used, HER2 being a surface receptor that is overexpressed in several tumor cells. Subsequently, the in vitro formation of a PC has been developed using fetal bovine serum supplemented with human fibrinogen. Dynamic Light Scattering (DLS), Nanoparticle Tracking Analysis (NTA), Laser Doppler Electrophoresis (LDE), and Gel Chromatography techniques have been used to assure a complete physico-chemical characterization of the nano-complexes with (LLNCs-αHER2-PC) and without (LLNCs-αHER2) the surrounding PC. In addition, cellular assays were performed to study the cellular uptake and the specific cellular-nanocarrier interactions using the SKBR3 (high expression of HER2) breast cancer cell line and human dermal fibroblasts (HDFa) (healthy cell line without expression of HER2 receptors as control), showing that the SKBR3 cell line had a higher transport rate (50-fold) than HDFa at 60 min with LLNCs-αHER2. Moreover, the SKBR3 cell line incubated with LLNCs-αHER2-PC suffered a significant reduction (40%) in the uptake. These results suggest that the formation of a PC onto LLNCs does not prevent specific cell targeting, although it does have an important influence on cell uptake.

Published
2023
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