1,013 results
Search Results
2. Determination of cyclic nucleotide-dependent protein kinase substrate specificity by the use of peptide libraries on cellulose paper
- Author
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Wolfgang R. Dostmann, Werner Tegge, Ronald Frank, and Franz Hofmann
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Paper ,Molecular Sequence Data ,Peptide ,Biochemistry ,chemistry.chemical_compound ,Cyclic nucleotide ,Structure-Activity Relationship ,Adenosine Triphosphate ,Escherichia coli ,Histone octamer ,Amino Acid Sequence ,Cellulose ,Phosphorylation ,Protein kinase A ,Cyclic GMP ,chemistry.chemical_classification ,Kinase ,Cyclic AMP-Dependent Protein Kinases ,Recombinant Proteins ,Amino acid ,Kinetics ,chemistry ,Oligopeptides ,Protein Kinases - Abstract
An iterative approach to the a priori determination of the substrate specificity of cAMP- and cGMP-dependent protein kinases (PKA and PKG) by the use of peptide libraries on cellulose paper is described. The starting point of the investigation was an octamer library with the general structure Ac-XXX12XXX, where X represents mixtures of all 20 natural amino acids and 1 and 2 represent individual amino acid residues. The library thus contained all possible 2.56 x 10(10) octamers, divided into 400 sublibraries with defined amino acids 1 and 2 each consisting of 6.4 x 10(7) sequences. After phosphorylation with the kinases in the presence of [gamma-32P]ATP, the sublibrarys Ac-XXXRRXXX and Ac-XXXRKXXX were identified as the best substrates for PKA and PKG, respectively. The second-generation libraries had the structures Ac-XXXRR12X and Ac-XXXRK12X for PKA and PKG and resulted in the most active sequence pools Ac-XXXRRASX and Ac-XXXRKKSX. After delineation of every position in the octameric sequence and extension of the investigation to decameric peptides, the best sequences, Ac-KRAERKASIY and Ac-TQKARKKSNA, were obtained for PKA and PKG, respectively. Promising octameric and decameric peptides were assembled 5 or 10 times each and assayed in order to determine the experimental scatter inherent in the approach. The kinetic data of several octameric and decameric sequences were determined in solution and compared to data for known substrates. The recognition motif of PKA was confirmed by this approach, and a novel substrate sequence for PKG was identified.(ABSTRACT TRUNCATED AT 250 WORDS)
- Published
- 1995
3. [Position paper of the Catalan Society of Gastroenterology: treatment of Genotype 1 Chronic Hepatitis C Virus with Triple Therapy]
- Author
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Miquel, Bruguera, Rafael, Esteban, Xavier, Forns, Ramón, Planas, Juan Carlos, Quer, Ricard, Solà, and Mercè, Vergara
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Diagnostic Imaging ,Liver Cirrhosis ,Patient Care Team ,Informed Consent ,Genotype ,Proline ,Biopsy ,Contraindications ,Interleukins ,Hepacivirus ,Hepatitis C, Chronic ,Viral Load ,Antiviral Agents ,Health Services Accessibility ,Polyethylene Glycols ,Recurrence ,Interferon Type I ,Ribavirin ,Humans ,Drug Therapy, Combination ,Protease Inhibitors ,Interferons ,Oligopeptides - Published
- 2012
4. Paper electrophoresis and chromatography of oligopeptides with N-terminal methionine
- Author
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Ronald G. Crystal, Norton A. Elson, and W. French Anderson
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Oligopeptide ,Methionine ,Chromatography ,Resolution (mass spectrometry) ,Chromatography, Paper ,Chemistry ,Biophysics ,Cell Biology ,Paper electrophoresis ,Tripeptide ,Biochemistry ,chemistry.chemical_compound ,Electrophoresis ,Sulfur Isotopes ,Small peptide ,Methods ,Protein biosynthesis ,Electrophoresis, Paper ,Amino Acid Sequence ,Amino Acids ,Oligopeptides ,Oxidation-Reduction ,Molecular Biology - Abstract
Several electrophoretic and chromatographic systems are described for the separation of di- and tripeptides with N-terminal methionine. The most effective methods of separation are electrophoresis in barbital-triethylamine-acetic acid, pH 8.0 and chromatography in n -butanol-acetic acid-water, 18:2:5. Increased resolution may be obtained by using two or more systems in combination. These methods were intended to separate and identify the oligopeptides formed during the initiation of eukaryotic protein synthesis, but may also be applied to the separation of small peptides generally.
- Published
- 1973
5. Paper electrophoresis of amino-acids and oligopeptides at very high potential gradients
- Author
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Gross D
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Electrophoresis ,Multidisciplinary ,Chromatography ,Materials science ,Paper electrophoresis ,Reduction (complexity) ,Heat generation ,Humans ,Electrophoresis, Paper ,Diffusion (business) ,Amino Acids ,Biological system ,Peptides ,Oligopeptides ,High potential - Abstract
THE importance of the amino-acids and oligopeptides and the complexity of their mixtures often encountered stimulate a demand for more powerful analytical tools to complement those already available. Paper electrophoresis does not show satisfactory resolving power, particularly in the separation of the neutral amino-acids, at the low potential gradients (5–10 V./cm.) usually applied. Its value is greatly enhanced, however, by the application of high potential gradients, most likely because the great reduction in time required for the molecules to traverse a given distance minimizes the detrimental effects of diffusion and moisture changes in the paper strip. Technical difficulties arising from increased heat generation can be overcome by modification of the design of basic equipment and by efficient cooling.
- Published
- 1955
6. Filter-paper ionophoresis of cupric complexes of neutral amino acids and oligopeptides
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P. R. Carnegie and R. L. M. Synge
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chemistry.chemical_classification ,History ,Oligopeptide ,Filter paper ,Stereochemistry ,Chemistry ,Articles ,Computer Science Applications ,Education ,Amino acid ,Neutral Amino Acids ,Amino Acids, Neutral ,Biochemistry ,Glycine ,Amino Acids ,Peptides ,Oligopeptides ,Copper - Published
- 1961
7. Merging Natural Products: Muraymycin–Sansanmycin Hybrid Structures as Novel Scaffolds for Potential Antibacterial Agents
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Christian Ducho, Giuliana Niro, and Stefanie Christine Weck
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structure–activity relationship ,natural products ,medicine.drug_class ,Antibiotics ,Transferases (Other Substituted Phosphate Groups) ,Microbial Sensitivity Tests ,Gram-Positive Bacteria ,010402 general chemistry ,01 natural sciences ,antibiotics ,Catalysis ,chemistry.chemical_compound ,Bacterial Proteins ,Transferases ,Gram-Negative Bacteria ,medicine ,Structure–activity relationship ,Uridine ,Integral membrane protein ,Biological Products ,Full Paper ,biology ,010405 organic chemistry ,Organic Chemistry ,General Chemistry ,Full Papers ,Antimicrobial ,biology.organism_classification ,Anti-Bacterial Agents ,0104 chemical sciences ,chemistry ,Biochemistry ,Antibacterial Agents ,Peptidoglycan ,Antibacterial activity ,Oligopeptides ,hybrid structures ,Nucleoside ,Bacteria ,nucleosides - Abstract
To overcome bacterial resistances, the need for novel antimicrobial agents is urgent. The class of so‐called nucleoside antibiotics furnishes promising candidates for the development of new antibiotics, as these compounds block a clinically unexploited bacterial target: the integral membrane protein MraY, a key enzyme in cell wall (peptidoglycan) biosynthesis. Nucleoside antibiotics exhibit remarkable structural diversity besides their uridine‐derived core motifs. Some sub‐classes also show specific selectivities towards different Gram‐positive and Gram‐negative bacteria, which are poorly understood so far. Herein, the synthesis of a novel hybrid structure is reported, derived from the 5′‐defunctionalized uridine core moiety of muraymycins and the peptide chain of sansanmycin B, as a new scaffold for the development of antimicrobial agents. The reported muraymycin–sansanmycin hybrid scaffold showed nanomolar activity against the bacterial target enzyme MraY, but displayed no significant antibacterial activity against S. aureus, E. coli, and P. aeruginosa., Two become one: Nucleoside antibiotics are natural products that show remarkable structural diversity. Herein, the synthesis of a novel non‐natural hybrid structure is reported, derived from the simplified uridine core moiety of muraymycins and the peptide chain of sansanmycin B. This muraymycin–sansanmycin hybrid scaffold showed nanomolar activity against the bacterial target enzyme MraY, thus suggesting it may be used for the development of antibacterial agents.
- Published
- 2020
8. Tumor targeting nanoparticle E749-57-HSP110-RGD elicits potent anti-tumor immune response in a CD8-dependent manner in cervical cancer-bearing mouse model
- Author
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Jun Tang, Yue Zhang, Tao Jing, Faliang Ren, and Bing Ni
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Papillomavirus E7 Proteins ,T cell ,medicine.medical_treatment ,Immunology ,Uterine Cervical Neoplasms ,CD8-Positive T-Lymphocytes ,Cancer Vaccines ,Epitope ,Mice ,Immune system ,Heat shock protein ,medicine ,Animals ,Humans ,Immunology and Allergy ,Cytotoxicity ,Pharmacology ,Expression vector ,Chemistry ,Immunity ,Immunotherapy ,Fusion protein ,Cell biology ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Nanoparticles ,Female ,Oligopeptides ,Research Paper - Abstract
Our previous research verified that HSP (heat shock protein) 110 could enhance the anti-tumor effect of HPV16 E7(49-57) epitope. In this study, to optimize the immunotherapy of this vaccine type, we developed and evaluated the anti-tumor immunity of a nanoparticle vaccine format assembling with E7(49-57)-HSP110 fusion expression plasmid and RGD-GGG-K(18) polypeptide. The nanoparticle vaccine was self-assembled from positively charged RGD-GGG-K(18) polypeptide and negatively charged fusion expression plasmid pIRES2-3× E7-HSP110-EGFP. The particle size, stability, expression of E7(49-57)-HSP110 fusion protein and the target ability of nanoparticle were determined, respectively. Specific CTL responses were determined by E7 tetramer staining and cytotoxicity assay in TC-1 tumor-bearing mice (CD4/CD8 knockout). The preventive and therapeutic experiments of nanoparticle vaccine were investigated in TC-1 tumor-bearing mice. Results showed that the RGD-GGG-K(18) polypeptide and pIRES2-3× E7-HSP110-EGFP plasmid self-assembled nanoparticles about 100 nanometers in diameter when the charge ratios of peptide/plasmid were 2. The nanoparticles effectively entered TC-1 cells directed by RGD target-peptide, and correctly expressed the E7-HSP110 fusion protein. The HSP110 effectively facilitated nanoparticles activating CD8(+)T cells than nanoparticles without HSP110, including the CD8(+) T cell number and the IFN-γ level; in contrast, the CD4(+)T cells immune response remained indiscriminate among the mice groups. This nanoparticle formulation inhibited tumor growth and prolonged the survival duration in the prophylactic and therapeutic mouse models. Therefore, the RGD-based tumor-targeting nanoparticle expressing E7(49-57)-HSP110 fusion protein can efficiently evoke anti-tumor activity and thus suggests it might be a favorable candidate for cervical cancer immunotherapy.
- Published
- 2021
9. Improving Antibody‐Tubulysin Conjugates through Linker Chemistry and Site‐Specific Conjugation
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Martha Anderson, Jamie A. Mitchell, Julia H. Cochran, Scott C. Jeffrey, Kim K. Emmerton, Joseph Z. Hamilton, Ivan Stone, Thomas A. Pires, Patrick J. Burke, Margo Zaval, Peter D. Senter, Robert P. Lyon, Steven Jin, and Andrew B. Waight
- Subjects
Immunoconjugates ,01 natural sciences ,Biochemistry ,Antibodies ,Rats, Sprague-Dawley ,Mice ,chemistry.chemical_compound ,Very Important Paper ,In vivo ,Drug Discovery ,Animals ,Humans ,cancer ,Potency ,General Pharmacology, Toxicology and Pharmaceutics ,Pharmacology ,Dipeptide ,Molecular Structure ,010405 organic chemistry ,Chemistry ,Communication ,Organic Chemistry ,glucuronides ,Xenograft Model Antitumor Assays ,Communications ,Rats ,0104 chemical sciences ,010404 medicinal & biomolecular chemistry ,Cell culture ,drug delivery ,Drug delivery ,Molecular Medicine ,Glucuronide ,Oligopeptides ,tubulysin ,Linker ,Conjugate - Abstract
Tubulysins have emerged in recent years as a compelling drug class for delivery to tumor cells via antibodies. The ability of this drug class to exert bystander activity while retaining potency against multidrug‐resistant cell lines differentiates them from other microtubule‐disrupting agents. Tubulysin M, a synthetic analogue, has proven to be active and well tolerated as an antibody‐drug conjugate (ADC) payload, but has the liability of being susceptible to acetate hydrolysis at the C11 position, leading to attenuated potency. In this work, we examine the ability of the drug‐linker and conjugation site to preserve acetate stability. Our findings show that, in contrast to a more conventional protease‐cleavable dipeptide linker, the β‐glucuronidase‐cleavable glucuronide linker protects against acetate hydrolysis and improves ADC activity in vivo. In addition, site‐specific conjugation can positively impact both acetate stability and in vivo activity. Together, these findings provide the basis for a highly optimized delivery strategy for tubulysin M., Stability and activity: Antibody‐tubulysin conjugates were evaluated for activity and acetate stability as a function of linker chemistry and conjugation site. Results show the glucuronide linker and S239 engineered cysteine conjugation preserve the acetate in circulation and improve in vivo activity relative to a dipeptide linker and endogenous cysteine conjugation.
- Published
- 2021
10. The isolated La-module of LARP1 mediates 3’ poly(A) protection and mRNA stabilization, dependent on its intrinsic PAM2 binding to PABPC1
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Kalle Gehring, Guennadi Kozlov, Richard J. Maraia, Bruno D. Fonseca, Sergei Gaidamakov, Sandy Mattijssen, and Amitabh Ranjan
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Polyadenylation ,RNA Stability ,Protein domain ,Biology ,Autoantigens ,Poly(A)-Binding Protein I ,Poly(A)-Binding Proteins ,03 medical and health sciences ,0302 clinical medicine ,PABPC1 ,Humans ,Protein Interaction Domains and Motifs ,RNA, Messenger ,Nucleotide Motifs ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,Binding Sites ,RNA recognition motif ,fungi ,Cell Biology ,MRNA stabilization ,Toll-Like Receptor 2 ,LARP1 ,Cell biology ,HEK293 Cells ,Ribonucleoproteins ,Toll-Like Receptor 9 ,030220 oncology & carcinogenesis ,Oligopeptides ,Linker ,Protein Binding ,Research Paper - Abstract
The protein domain arrangement known as the La-module, comprised of a La motif (LaM) followed by a linker and RNA recognition motif (RRM), is found in seven La-related proteins: LARP1, LARP1B, LARP3 (La protein), LARP4, LARP4B, LARP6, and LARP7 in humans. Several LARPs have been characterized for their distinct activity in a specific aspect of RNA metabolism. The La-modules vary among the LARPs in linker length and RRM subtype. The La-modules of La protein and LARP7 bind and protect nuclear RNAs with UUU-3ʹ tails from degradation by 3ʹ exonucleases. LARP4 is an mRNA poly(A) stabilization factor that binds poly(A) and the cytoplasmic poly(A)-binding protein PABPC1 (also known as PABP). LARP1 exhibits poly(A) length protection and mRNA stabilization similar to LARP4. Here, we show that these LARP1 activities are mediated by its La-module and dependent on a PAM2 motif that binds PABP. The isolated La-module of LARP1 is sufficient for PABP-dependent poly(A) length protection and mRNA stabilization in HEK293 cells. A point mutation in the PAM2 motif in the La-module impairs mRNA stabilization and PABP binding in vivo but does not impair oligo(A) RNA binding by the purified recombinant La-module in vitro. We characterize the unusual PAM2 sequence of LARP1 and show it may differentially affect stable and unstable mRNAs. The unique LARP1 La-module can function as an autonomous factor to confer poly(A) protection and stabilization to heterologous mRNAs.
- Published
- 2020
11. δ-opioid receptor activation protects against Parkinson’s disease-related mitochondrial dysfunction by enhancing PINK1/Parkin-dependent mitophagy
- Author
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Jiahao Mao, Ya Peng, Ying Xia, Yilin Yang, Feng Zhi, Naiyuan Shao, Gianfranco Balboni, and Yuan Xu
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1-Methyl-4-phenylpyridinium ,Aging ,Ubiquitin-Protein Ligases ,PINK1 ,Mitochondrion ,PC12 Cells ,Neuroprotection ,Parkin ,δ-opioid receptor ,Parkinsonian Disorders ,Superoxides ,Receptors, Opioid, delta ,Mitophagy ,medicine ,Animals ,Membrane Potential, Mitochondrial ,Gene knockdown ,δ-Opioid receptor ,Herbicides ,Chemistry ,Cell Biology ,Hypoxia (medical) ,Cell Hypoxia ,Mitochondria ,Rats ,Up-Regulation ,Cell biology ,Gene Knockdown Techniques ,Benzimidazoles ,medicine.symptom ,Oligopeptides ,Protein Kinases ,Research Paper - Abstract
Our previous studies have shown that the δ-opioid receptor (DOR) is an important neuroprotector via the regulation of PTEN-induced kinase 1 (PINK1), a mitochondria-related molecule, under hypoxic and MPP+ insults. Since mitochondrial dysfunctions are observed in both hypoxia and MPP+ insults, this study further investigated whether DOR is cytoprotective against these insults by targeting mitochondria. Through comparing DOR-induced responses to hypoxia versus MPP+-induced parkinsonian insult in PC12 cells, we found that both hypoxia and MPP+ caused a collapse of mitochondrial membrane potential and severe mitochondrial dysfunction. In sharp contrast to its inappreciable effect on mitochondria in hypoxic conditions, DOR activation with UFP-512, a specific agonist, significantly attenuated the MPP+-induced mitochondrial injury. Mechanistically, DOR activation effectively upregulated PINK1 expression and promoted Parkin’s mitochondrial translocation and modification, thus enhancing the PINK1-Parkin mediated mitophagy. Either PINK1 knockdown or DOR knockdown largely interfered with the DOR-mediated mitoprotection in MPP+ conditions. Moreover, there was a major difference between hypoxia versus MPP+ in terms of the regulation of mitophagy with hypoxia-induced mitophagy being independent from DOR-PINK1 signaling. Taken together, our novel data suggest that DOR activation is neuroprotective against parkinsonian injury by specifically promoting mitophagy in a PINK1-dependent pathway and thus attenuating mitochondrial damage.
- Published
- 2020
12. Identification and validation of a novel anti-virulent that binds to pyoverdine and inhibits its function
- Author
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Alexey V. Revtovich, Natalia V. Kirienko, Quinn Kleerekoper, Xu Wang, and Donghoon Kang
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Microbiology (medical) ,in silico structure modeling ,Virulence Factors ,medicine.drug_class ,Immunology ,Antibiotics ,Virulence ,Bronchi ,Infectious and parasitic diseases ,RC109-216 ,Molecular Dynamics Simulation ,Biology ,medicine.disease_cause ,Microbiology ,structure-activity relationship (SAR) ,Structure-Activity Relationship ,03 medical and health sciences ,chemistry.chemical_compound ,Immune system ,Antibiotic resistance ,medicine ,Animals ,Humans ,Computer Simulation ,In patient ,Pseudomonas aeruginosa (P. aeruginosa) ,solution nuclear magnetic resonance (NMR) ,Caenorhabditis elegans ,030304 developmental biology ,0303 health sciences ,Pyoverdine ,pyoverdine ,030306 microbiology ,Pseudomonas aeruginosa ,Epithelial Cells ,Anti-Bacterial Agents ,High-Throughput Screening Assays ,Molecular Docking Simulation ,Infectious Diseases ,chemistry ,Parasitology ,Oligopeptides ,Function (biology) ,Research Article ,Research Paper - Abstract
Pseudomonas aeruginosa: causes serious infections in patients with compromised immune systems and exhibits resistance to multiple antibiotics. The rising threat of antimicrobial resistance means that new methods are necessary for treating microbial infections. We conducted a high-throughput screen for compounds that can quench the innate fluorescence of the chromophore region of the P. aeruginosa siderophore pyoverdine, a key virulence factor. Several hits were identified that effectively quench pyoverdine fluorescence, and two compounds considerably improved the survival of Caenorhabditis elegans when worms were challenged with P. aeruginosa. Commercially available analogs of the best hit, PQ3, were tested for their ability to rescue C. elegans from P. aeruginosa and to interact with pyoverdine via fluorescence and solution NMR spectroscopy. 1H-15N and 1H-13C HSQC NMR were used to identify the binding site of PQ3c. The structure model of pyoverdine in complex with PQ3c was obtained using molecular docking and molecular dynamics simulations. PQ3c occupied a shallow groove on pyoverdine formed by the chromophore and N-terminal residues of the peptide chain. Electrostatic interactions and π-orbital stacking drove stabilization of this binding. PQ3c may serve as a scaffold for the development of pyoverdine inhibitors with higher potency and specificity. The discovery of a small-molecule binding site on apo-pyoverdine with functional significance provides a new direction in the search of therapeutically effective reagent to treat P. aeruginosa infections. Abbreviations: NMR: nuclear magnetic resonance; SAR: structure–activity relationship; MD: molecular dynamics; RMSF: root-mean-square fluctuation; HSQC: heteronuclear single quantum correlation; DMSO: dimethyl sulfoxide; Δδavg: average amide chemical shift change; DSS: 2,2-dimethyl-2-silapentane-5-sulfonate; RMSD: root-mean-square deviation; LJ-SR: Lennard–Jones short-range; Coul-SR: Coulombic short-range; FRET: fluorescence resonance energy transfer
- Published
- 2020
13. BNIP3L/NIX degradation leads to mitophagy deficiency in ischemic brains
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Wei-dong Tang, Wanqing Zheng, Yue Li, Zheng-Hong Qin, Xiaoli Wu, Feng Han, Weiwei Hu, Yanrong Zheng, Ming Cao, Mengru Liu, Liang Fang, Shijia Ma, Zhong Chen, Lei Jiang, Xiangnan Zhang, Jiaying Wu, and Wenping Yan
- Subjects
0301 basic medicine ,Ischemia ,Biology ,Mitochondrial Proteins ,03 medical and health sciences ,chemistry.chemical_compound ,Sequestosome 1 ,Mitophagy ,MG132 ,Autophagy ,medicine ,Animals ,education ,Molecular Biology ,Mice, Inbred BALB C ,education.field_of_study ,030102 biochemistry & molecular biology ,Membrane Proteins ,Cell Biology ,medicine.disease ,Mitochondria ,Cell biology ,Mice, Inbred C57BL ,030104 developmental biology ,Proteasome ,chemistry ,Proteasome inhibitor ,Apoptosis Regulatory Proteins ,Reactive Oxygen Species ,Oligopeptides ,MAP1LC3B ,Research Paper ,medicine.drug - Abstract
Mitophagy, the elimination of damaged mitochondria through autophagy, promotes neuronal survival in cerebral ischemia. Previous studies found deficient mitophagy in ischemic neurons, but the mechanisms are still largely unknown. We determined that BNIP3L/NIX, a mitophagy receptor, was degraded by proteasomes, which led to mitophagy deficiency in both ischemic neurons and brains. BNIP3L exists as a monomer and homodimer in mammalian cells, but the effects of homodimer and monomer on mitophagy are unclear. Site-specific mutations in the transmembrane domain of BNIP3L (S195A and G203A) only formed the BNIP3L monomer and failed to induce mitophagy. Moreover, overexpression of wild-type BNIP3L, in contrast to the monomeric BNIP3L, rescued the mitophagy deficiency and protected against cerebral ischemic injury. The macroautophagy/autophagy inhibitor 3-MA and the proteasome inhibitor MG132 were used in cerebral ischemic brains to identify how BNIP3L was reduced. We found that MG132 blocked the loss of BNIP3L and subsequently promoted mitophagy in ischemic brains. In addition, the dimeric form of BNIP3L was more prone to be degraded than its monomeric form. Carfilzomib, a drug for multiple myeloma therapy that inhibits proteasomes, reversed the BNIP3L degradation and restored mitophagy in ischemic brains. This treatment protected against either acute or chronic ischemic brain injury. Remarkably, these effects of carfilzomib were abolished in bnip3l(-/-) mice. Taken together, the present study linked BNIP3L degradation by proteasomes with mitophagy deficiency in cerebral ischemia. We propose carfilzomib as a novel therapy to rescue ischemic brain injury by preventing BNIP3L degradation. Abbreviations: 3-MA: 3-methyladenine; AAV: adeno-associated virus; ATG7: autophagy related 7; BCL2L13: BCL2-like 13 (apoptosis facilitator); BNIP3L/NIX: BCL2/adenovirus E1B interacting protein 3-like; CCCP: carbonyl cyanide 3-chlorophenylhydrazone; CFZ: carfilzomib; COX4I1: cytochrome c oxidase subunit 4I1; CQ: chloroquine; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GFP: green fluorescent protein; I-R: ischemia-reperfusion; MAP1LC3A/LC3A: microtube-associated protein 1 light chain 3 alpha; MAP1LC3B/LC3B: microtube-associated protein 1 light chain 3 beta; O-R: oxygen and glucose deprivation-reperfusion; OGD: oxygen and glucose deprivation; PHB2: prohibitin 2; pMCAO: permanent middle cerebral artery occlusion; PRKN/PARK2: parkin RBR E3 ubiquitin protein ligase; PT: photothrombosis; SQSTM1: sequestosome 1; tMCAO: transient middle cerebral artery occlusion; TOMM20: translocase of outer mitochondrial membrane 20; TTC: 2,3,5-triphenyltetrazolium hydrochloride.
- Published
- 2020
14. Oligopeptide-modified poly(beta-amino ester)s-coated AdNuPARmE1A: Boosting the efficacy of intravenously administered therapeutic adenoviruses
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Cristina Fornaguera, Maria Rovira-Rigau, Cristina Castells-Sala, Anna Cascante, Cristina Fillat, Salvador Borrós, Miguel Ángel Lázaro, Pau Brugada-Vilà, Lorenzo Albertazzi, Molecular Biosensing for Med. Diagnostics, and ICMS Core
- Subjects
Oncolytic adenovirus ,Biodistribution ,Polymers ,Population ,Medicine (miscellaneous) ,Antineoplastic Agents ,02 engineering and technology ,Pharmacology ,SDG 3 – Goede gezondheid en welzijn ,Mice ,03 medical and health sciences ,Systemic delivery ,SDG 3 - Good Health and Well-being ,In vivo ,Cell Line, Tumor ,Neoplasms ,Animals ,Humans ,education ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,030304 developmental biology ,Oncolytic Virotherapy ,0303 health sciences ,education.field_of_study ,Chemistry ,Immunogenicity ,Pancreatic cancer ,021001 nanoscience & nanotechnology ,In vitro ,Oncolytic virus ,Mice, Inbred C57BL ,Oncolytic Viruses ,HEK293 Cells ,RAW 264.7 Cells ,Cancer cell ,Poly(β-amino ester)s ,Polymer-coated viral vectors ,0210 nano-technology ,Oligopeptides ,Research Paper - Abstract
Oncolytic adenoviruses are used as agents for the treatment of cancer. However, their potential is limited due to the high seroprevalence of anti-adenovirus neutralizing antibodies (nAbs) within the population and the rapid liver sequestration when systemically administered. To overcome these challenges, we explored using nanoparticle formulation to boost the efficacy of systemic oncolytic adenovirus administration. Methods: Adenovirus were conjugated with PEGylated oligopeptide-modified poly(β-amino ester)s (OM-pBAEs). The resulting coated viral formulation was characterized in terms of surface charge, size, aggregation state and morphology and tested for anti-adenovirus nAbs evasion and activity in cancer cells. In vivo pharmacokinetics, biodistribution, tumor targeting, and immunogenicity studies were performed. The antitumor efficacy of the oncolytic adenovirus AdNuPARmE1A coated with OM-pBAEs (SAG101) in the presence of nAbs was evaluated in pancreatic ductal adenocarcinoma (PDAC) mouse models. Toxicity of the coated formulation was analyzed in vivo in immunocompetent mice. Results: OM-pBAEs conjugated to adenovirus and generated discrete nanoparticles with a neutral charge and an optimal size. The polymeric coating with the reporter AdGFPLuc (CPEG) showed enhanced transduction and evasion of antibody neutralization in vitro. Moreover, systemic intravenous administration of the formulation showed improved blood circulation and reduced liver sequestration, substantially avoiding activation of nAb production. OM-pBAEs coating of the oncolytic adenovirus AdNuPARmE1A (SAG101) improved its oncolytic activity in vitro and enhanced antitumor efficacy in PDAC mouse models. The coated formulation protected virions from neutralization by nAbs, as antitumor efficacy was preserved in their presence but was completely lost in mice that received the non-formulated AdNuPARmE1A. Finally, coated-AdNuPARmE1A showed reduced toxicity when high doses of the formulation were administered. Conclusions: The developed technology represents a promising improvement for future clinical cancer therapy using oncolytic adenoviruses.
- Published
- 2020
15. Near-infrared fluorescence imaging-guided focused ultrasound-mediated therapy against Rheumatoid Arthritis by MTX-ICG-loaded iRGD-modified echogenic liposomes
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Hongmei Liu, Mei-Jun Zhou, Fei Yan, Haohan Wu, Hao Wu, Yanni He, Ran Xiong, and Zhili Xu
- Subjects
Male ,Near-Infrared Fluorescence Imaging ,Fluorescence-lifetime imaging microscopy ,Swine ,Angiogenesis ,Ultrasonic Therapy ,medicine.medical_treatment ,Medicine (miscellaneous) ,02 engineering and technology ,01 natural sciences ,Targeted therapy ,Echogenic liposomes ,Arthritis, Rheumatoid ,Mice ,chemistry.chemical_compound ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,Spectroscopy, Near-Infrared ,Optical Imaging ,021001 nanoscience & nanotechnology ,iRGD ,Ultrasonic Waves ,Rheumatoid arthritis ,0210 nano-technology ,Oligopeptides ,Research Paper ,medicine.drug ,Indocyanine Green ,musculoskeletal diseases ,Near-infrared fluorescence imaging ,010402 general chemistry ,Cell Line ,Ultrasound-controlled drug release ,Synovitis ,Human Umbilical Vein Endothelial Cells ,medicine ,Animals ,Humans ,Fluorescent Dyes ,business.industry ,Endothelial Cells ,Integrin alphaVbeta3 ,medicine.disease ,0104 chemical sciences ,Methotrexate ,RAW 264.7 Cells ,chemistry ,Liposomes ,Cancer research ,business ,Indocyanine green - Abstract
Rheumatoid arthritis (RA), a common inflammatory disorder of the joints characterized by synovitis and pannus formation, often results in irreversible joint erosion and disability. Methotrexate (MTX) is the first-line drug against RA, but the therapeutic effects are sub-optimal due to its poor retention at the target sites and systemic side effects. Multifunctional nanoparticles are highly promising agents for minimally invasive, traceable and effective targeted therapy. Methods: This study developed iRGD peptide-functionalized echogenic liposomes (iELPs) which encapsulates MTX and indocyanine green (ICG) fluorescent probe through the thin film-hydration method. Results: The resulting iELPs showed high affinity for endothelial cells overexpressing αvβ3 integrin, favorable acoustic response and fluorescence tracking properties. Also, near-infrared (NIR) fluorescence imaging of iELPs and ultrasound-triggered drug release of MTX were proved in a mouse RA model, greatly improving the therapeutic efficacy and reducing MTX side effects. Histological assessment of the articular tissues further revealed significantly lower inflammatory cell infiltration and angiogenesis in the iELPs-treated and sonicated mice. Conclusion: Our study provides a promising nanoplatform for integrating ultrasound-controlled drug release and NIR fluorescence imaging for RA treatment.
- Published
- 2020
16. A simple screening method for detecting bindings between oligopeptides and HLA-DR molecules on filter papers: Possible application for mapping of putative helper T-cell epitopes on MSP1 of Plasmodium falciparum
- Author
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Judson L. Leafasia, Karen Igarashi, Akira Ishii, Nobuo Ohta, Takashi Suzuki, Mariko Hato, Jun Fu, Yasuo Chinzei, and Hiroyuki Matsuoka
- Subjects
Immunology ,Molecular Sequence Data ,Plasmodium falciparum ,Epitopes, T-Lymphocyte ,Peptide ,Lymphocyte Activation ,Microbiology ,Epitope ,Antigen ,Virology ,HLA-DR ,Animals ,Humans ,Amino Acid Sequence ,Binding site ,Peptide sequence ,Merozoite Surface Protein 1 ,chemistry.chemical_classification ,biology ,HLA-DR Antigens ,T-Lymphocytes, Helper-Inducer ,biology.organism_classification ,Molecular biology ,Epitope mapping ,chemistry ,Immunologic Techniques ,Oligopeptides ,Epitope Mapping - Abstract
Binding capacities of synthetic peptides to HLA-DR molecules were tested on filter papers to identify putative helper T-cell epitopes on a malarial protein. The antigen tested was the merozoite surface glycoprotein 1 (MSP1) of Plasmodium falciparum, a vaccine candidate targeting the asexual erythrocytic stage. Bindings between synthetic oligopeptides and HLA-DR molecules were tested. Such bindings were not non-specific, and a known helper T-cell epitope peptide showed positive binding to the restricting HLA-DR molecule. By using this screening system, we observed the unequal distribution of HLA-DR-binding peptides in 10 out of 17 MSP1 blocks tested. Block #6 of MSP1 seemed to show the highest frequency in the positive binding; on the other hand, blocks #1 and #17, both of which were thought to be vaccine candidate regions, contained fewer HLA-DR binding peptides. This was not inconsistent with the results that block #17 was less stimulatory to peripheral T cells than block #6. The peptides with positive binding to HLA-DR showed actual epitope activities when we tested peptide-driven proliferation of human bulk T-cell lines, and association between the two parameters was statistically significant (P
17. Targeted Discovery of Tetrapeptides and Cyclic Polyketide-Peptide Hybrids from a Fungal Antagonist of Farming Termites
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Soohyun Um, Helmar Görls, Z. Wilhelm de Beer, Huijuan Guo, Nina B. Kreuzenbeck, Felix Schalk, and Christine Beemelmanns
- Subjects
Models, Molecular ,Stereochemistry ,natural products ,polyketide synthases ,Molecular Conformation ,Peptide ,Fungus ,010402 general chemistry ,Tandem mass spectrometry ,Crystallography, X-Ray ,01 natural sciences ,Biochemistry ,Peptides, Cyclic ,Polyketide ,Metabolomics ,Nonribosomal peptide ,Drug Discovery ,Bioassay ,Molecular Biology ,Nuclear Magnetic Resonance, Biomolecular ,symbionts ,chemistry.chemical_classification ,Biological Products ,biology ,Full Paper ,010405 organic chemistry ,Organic Chemistry ,Stereoisomerism ,Termitomyces ,Full Papers ,biology.organism_classification ,metabolomics ,0104 chemical sciences ,chemistry ,Polyketides ,peptides ,Molecular Medicine ,Two-dimensional nuclear magnetic resonance spectroscopy ,Oligopeptides ,nonribosomal peptide synthetase - Abstract
Herein, we report the targeted isolation and characterization of four linear nonribosomally synthesized tetrapeptides (pseudoxylaramide A–D) and two cyclic nonribosomal peptide synthetase‐polyketide synthase‐derived natural products (xylacremolide A and B) from the termite‐associated stowaway fungus Pseudoxylaria sp. X187. The fungal strain was prioritized for further metabolic analysis based on its taxonomical position and morphological and bioassay data. Metabolic data were dereplicated based on high‐resolution tandem mass spectrometry data and global molecular networking analysis. The structure of all six new natural products was elucidated based on a combination of 1D and 2D NMR analysis, Marfey's analysis and X‐ray crystallography., Chemical treasures of a fungal stowaway: Structural analysis of six new chemical features revealed four linear tetrapeptides and two cyclic NRPS‐polyketide‐derived natural products from the fungal antagonist Pseudoxylaria sp. X187 of fungus‐growing termites.
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- 2020
18. Reduction of Elevated Proton Leak Rejuvenates Mitochondria in the Aged Cardiomyocyte
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David J. Marcinek, Wang Wang, Peter S. Rabinovitch, Hazel H. Szeto, Huiliang Zhang, and Nathan N. Alder
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0301 basic medicine ,Leak ,Health (social science) ,Mouse ,cardiomyocyte ,Mitochondrion ,Health Professions (miscellaneous) ,Mitochondria, Heart ,Membrane Potentials ,Abstracts ,chemistry.chemical_compound ,Adenosine Triphosphate ,0302 clinical medicine ,Medicine ,Myocytes, Cardiac ,Biology (General) ,AcademicSubjects/SOC02600 ,Cells, Cultured ,Cellular Senescence ,0303 health sciences ,education.field_of_study ,SS-31 ,General Neuroscience ,MPTP ,030302 biochemistry & molecular biology ,Age Factors ,General Medicine ,Hydrogen-Ion Concentration ,3. Good health ,mitochondria ,Cardiology ,Protons ,Oligopeptides ,Research Article ,medicine.medical_specialty ,QH301-705.5 ,Science ,Population ,Diastole ,Mice, Transgenic ,General Biochemistry, Genetics and Molecular Biology ,Cardiac dysfunction ,03 medical and health sciences ,Internal medicine ,Animals ,Effective treatment ,Life-span and Life-course Studies ,education ,030304 developmental biology ,Session 3023 (Paper) ,General Immunology and Microbiology ,Mitochondrial Permeability Transition Pore ,business.industry ,aging ,Adenine Nucleotide Translocator 1 ,Cell Biology ,Elamipretide ,medicine.disease ,Rats, Inbred F344 ,Cardiovascular Health and Disease (paper) ,Mice, Inbred C57BL ,030104 developmental biology ,Endocrinology ,Mitochondrial permeability transition pore ,chemistry ,Heart failure ,Rat ,Energy Metabolism ,Reactive Oxygen Species ,business ,Heart failure with preserved ejection fraction ,030217 neurology & neurosurgery ,proton leak - Abstract
Aging-associated diseases, including cardiac dysfunction, are increasingly common in the population. However, the mechanisms of physiologic aging in general, and cardiac aging in particular, remain poorly understood. While effective medical interventions are available for some kinds of heart failure, one age-related impairment, diastolic dysfunction in Heart Failure with Preserved Ejection Fraction (HFpEF) is lacking a clinically effective treatment. Using the model of naturally aging mice and rats, we show direct evidence of increased proton leak in the aged heart mitochondria. Moreover, we identified ANT1 as mediating the increased proton permeability of old cardiomyocytes. Most importantly, the tetra-peptide drug SS-31 (elamipretide) prevents age-related excess proton entry, decreases the mitochondrial flash activity and mitochondrial permeability transition pore (mPTP) opening and rejuvenates mitochondrial function by direct association with ANT1 and the mitochondrial ATP synthasome. Our results uncover a novel mechanism of age-related cardiac dysfunction and elucidate how SS-31 is able to reverse this clinically important complication of cardiac aging. Significance Aging is the greatest risk factor for cardiac dysfunction, including Heart Failure with Preserved Ejection Fraction (HFpEF). Unfortunately, the mechanisms of cardiac aging remain elusive, and there are no effective pharmacologic therapies for HFpEF. Here, we show direct evidence of increased proton leak in aged cardiac mitochondria and have identified ANT1 as mediating the increased proton permeability of old cardiomyocytes. Moreover, the mitochondrial-targeted tetra-peptide SS-31 (elamipretide) prevents the age-related excess proton entry and rejuvenates mitochondrial function by direct association with ANT1 and the mitochondrial ATP synthasome, resulting in alleviation of diastolic dysfunction in old mice. Our results unmask a novel mechanism of cardiac aging and elucidate how SS-31 reverses this clinically important complication of aging.
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- 2020
19. Paricalcitol accelerates BACE1 lysosomal degradation and inhibits calpain-1 dependent neuronal loss in APP/PS1 transgenic mice
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Zhan-You Wang, Jun-Lin Liu, Yong-Gang Fan, Tian Guo, Yu-Ting Cai, Xueshi Huang, Han Xue, Xiao-Ran Han, Chuang Guo, and Yan-Chun Li
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0301 basic medicine ,Paricalcitol ,Research paper ,Mitochondrion ,Calcitriol receptor ,Mice ,0302 clinical medicine ,Aspartic Acid Endopeptidases ,Neurons ,biology ,medicine.diagnostic_test ,Calpain ,Chemistry ,Brain ,General Medicine ,Alzheimer's disease ,LRP1 ,humanities ,Mitochondria ,Cell biology ,Lysosomal degradation ,8-Hydroxy-2'-Deoxyguanosine ,8-hydroxyguanosine ,030220 oncology & carcinogenesis ,Ergocalciferols ,Neuronal loss ,Oligopeptides ,Low Density Lipoprotein Receptor-Related Protein-1 ,medicine.drug ,Genetically modified mouse ,Mice, Transgenic ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,Western blot ,Alzheimer Disease ,mental disorders ,Presenilin-1 ,medicine ,Animals ,Humans ,Amyloid beta-Peptides ,β-Site APP cleavage enzyme 1 ,Disease Models, Animal ,030104 developmental biology ,Gene Expression Regulation ,Proteolysis ,biology.protein ,Amyloid Precursor Protein Secretases ,Lysosomes ,Immunostaining - Abstract
Background Recent studies have revealed that vitamin D deficiency may increase the risk of Alzheimer's disease, and vitamin D supplementation may be effective strategy to ameliorate the neurodegenerative process in Alzheimer's disease patients. Paricalcitol (PAL), a low-calcemic vitamin D receptor agonist, is clinically used to treat secondary hyperparathyroidism. However, the potential application of PAL for treating neurodegenerative disorders remains unexplored. Methods The APP/PS1 mice were intraperitoneally injected with PAL or vehicle every other day for 15 weeks. The β-amyloid (Aβ) production was confirmed using immunostaining and enzyme linked immunosorbent assay. The underlying mechanism was verified by western blot and immunostaining in vivo and in vitro. Findings Long-term PAL treatment clearly reduced β-amyloid (Aβ) generation and neuronal loss in APP/PS1 transgenic mouse brains. PAL stimulated the expression of low-density lipoprotein receptor-related protein 1 (LRP1) possibly through inhibiting sterol regulatory element binding protein-2 (SREBP2); PAL also promoted LRP1-mediated β-site APP cleavage enzyme 1 (BACE1) transport to late endosomes, thus increasing the lysosomal degradation of BACE1. Furthermore, PAL diminished 8-hydroxyguanosine (8-OHdG) generation in neuronal mitochondria via enhancing base excision repair (BER), resulting in the attenuation of calpain-1-mediated neuronal loss. Interpretation The present data demonstrate that PAL can reduce Aβ generation through accelerating BACE1 lysosomal degradation and can inhibit neuronal loss through suppressing mitochondrial 8-OHdG generation. Hence, PAL might be a promising agent for treating Alzheimer's disease. Fund This study was financially supported by the Natural Science Foundation of China (U1608282).
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- 2019
20. Orthogonally Stimulated Assembly/Disassembly of Depsipeptides by Rational Chemical Design
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Michaela Pieszka, Adriana Maria Sobota, Tanja Weil, David Y. W. Ng, and Jasmina Gačanin
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rearrangement ,Supramolecular chemistry ,010402 general chemistry ,01 natural sciences ,Biochemistry ,Serine ,Methionine ,Very Important Paper ,Amino Acid Sequence ,stimulus-responsive assembly ,Molecular Biology ,Peptide sequence ,amyloid peptides ,Depsipeptide ,Oligopeptide ,010405 organic chemistry ,Chemistry ,Communication ,fungi ,Organic Chemistry ,food and beverages ,Esters ,self-assembly ,Communications ,Nanostructures ,0104 chemical sciences ,Assembly disassembly ,Biophysics ,Molecular Medicine ,Protein Conformation, beta-Strand ,Self-assembly ,Oligopeptides ,Oxidation-Reduction ,Chemical design - Abstract
Controlling the assembly and disassembly of cross‐β‐sheet‐forming peptides is one of the predominant challenges for this class of supramolecular material. As they constitute a continuously propagating material, every atomic change can be exploited to bring about distinct responses at the architectural level. We report herein that, by using rational chemical design, serine and methionine can both be used as orthogonal chemical triggers to signal assembly/disassembly through their corresponding stimuli. Serine is used to construct an ester‐bond oligopeptide that can undergo O,N‐acyl rearrangement, whereas methionine is sensitive to oxidation by H2O2. Using the example peptide sequence, KIKISQINM, we demonstrate that assembly and disassembly can be independently controlled on demand.
- Published
- 2019
21. Montelukast enhances cytocidal effects of carfilzomib in multiple myeloma by inhibiting mTOR pathway
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Chao Wu, Jia Tong, Wenjun Yu, Wenbin Xu, Hua Yan, Qing Yu, and Ying-Li Wu
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Cyclopropanes ,0301 basic medicine ,Cancer Research ,Stromal cell ,Acetates ,Sulfides ,Transfection ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,immune system diseases ,Cell Line, Tumor ,Animals ,Humans ,Medicine ,Cytotoxic T cell ,Anti-Asthmatic Agents ,Cytotoxicity ,PI3K/AKT/mTOR pathway ,Montelukast ,Multiple myeloma ,Pharmacology ,business.industry ,TOR Serine-Threonine Kinases ,medicine.disease ,Xenograft Model Antitumor Assays ,Carfilzomib ,Disease Models, Animal ,030104 developmental biology ,medicine.anatomical_structure ,Oncology ,chemistry ,030220 oncology & carcinogenesis ,Quinolines ,Cancer research ,Molecular Medicine ,Bone marrow ,Multiple Myeloma ,business ,Oligopeptides ,Research Paper ,medicine.drug - Abstract
Montelukast is an anti-asthmatic medication, and has recently showed its inhibitory effects on the proliferation of cancers. The purpose of this study was to identify the cytotoxic effects of montelukast on multiple myeloma (MM) cells and the combination effects of montelukast and carfilzomib in the treatment of MM. Results revealed that montelukast induced a dose- and time-dependent cytotoxicity in MM cells lines and significantly suppressed the colony formation of myeloma cells. Furthermore, montelukast enhanced the cytotoxicity of carfilzomib in MM cell lines. This anti-tumor effect was associated with decreased c-Myc via the inhibition of mTOR signaling pathway. Moreover, the combination of montelukast and carfilzomib induced apoptosis of myeloma cells effectively, even in the presence of bone marrow stromal cells (BMSCs). It is more important to note that the co-treatment exhibited similar cytocidal effects in carfilzomib-resistant cell lines (U266R and 8226R). In addition, the combined effects were noted in two MM xenograft mice models and 7 cases of human CD138(+) myeloma cells (4 newly diagnosed cases and 3 relapsed cases) with no cytotoxicity on peripheral blood mononuclear cells (PBMCs) from 5 healthy donors. Our data suggested that montelukast enhanced the cytotoxicity of carfilzomib in both carfilzomib-sensitive and carfilzomib-resistant MM cell lines. These findings may facilitate the development of therapeutic strategies and provide a promising therapeutic combination regimen for the treatment of refractory myeloma.
- Published
- 2018
22. Impact of total variation regularized expectation maximization reconstruction on the image quality of
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Feng-Jiao, Yang, Shu-Yue, Ai, Runze, Wu, Yang, Lv, Hui-Fang, Xie, Yun, Dong, Qing-Le, Meng, and Feng, Wang
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Aged, 80 and over ,Male ,Full Paper ,Prostate ,Prostatic Neoplasms ,Reproducibility of Results ,Gallium Radioisotopes ,Middle Aged ,Positron Emission Tomography Computed Tomography ,Image Interpretation, Computer-Assisted ,Humans ,Oligopeptides ,Edetic Acid ,Gallium Isotopes ,Aged - Abstract
OBJECTIVES: To investigate the impact of total variation regularized expectation maximization (TVREM) reconstruction on the image quality of (68)Ga-PSMA-11 PET/CT using phantom and patient data. METHODS: Images of a phantom with small hot sphere inserts and 20 prostate cancer patients were acquired with a digital PET/CT using list-mode and reconstructed with ordered subset expectation maximization (OSEM) and TVREM with seven penalisation factors between 0.01 and 0.42 for 2 and 3 minutes-per-bed (m/b) acquisition. The contrast recovery (CR) and background variability (BV) of the phantom, image noise of the liver, and SUV(max) of the lesions were measured. Qualitative image quality was scored by two radiologists using a 5-point scale (1-poor, 5-excellent). RESULTS: The performance of CR, BV, and image noise, and the gain of SUV(max) was higher for TVREM 2 m/b groups with the penalization of 0.07 to 0.28 compared to OSEM 3 m/b group (all p < 0.05). The image noise of OSEM 3 m/b group was equivalent to TVREM 2 and 3 m/b groups with a penalization of 0.14 and 0.07, while lesions’ SUV(max) increased 15 and 20%. The highest qualitative score was attained at the penalization of 0.21 (3.30 ± 0.66) for TVREM 2 m/b groups and the penalization 0.14 (3.80 ± 0.41) for 3 m/b group that equal to or greater than OSEM 3 m/b group (2.90 ± 0.45, p = 0.2 and p < 0.001). CONCLUSIONS: TVREM improves lesion contrast and reduces image noise, which allows shorter acquisition with preserved image quality for PSMA PET/CT. ADVANCES IN KNOWLEDGE: TVREM reconstruction with optimized penalization factors can generate higher quality PSMA-PET images for prostate cancer diagnosis.
- Published
- 2021
23. Prostate cancer GTV delineation with biparametric MRI and
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Nathan, Hearn, John, Blazak, Philip, Vivian, Dinesh, Vignarajah, Katelyn, Cahill, Daisy, Atwell, Jim, Lagopoulos, and Myo, Min
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Aged, 80 and over ,Male ,Observer Variation ,Full Paper ,Prostate ,Prostatic Neoplasms ,Reproducibility of Results ,Gallium Radioisotopes ,Middle Aged ,Magnetic Resonance Imaging ,Positron-Emission Tomography ,Image Interpretation, Computer-Assisted ,Humans ,Radiopharmaceuticals ,Oligopeptides ,Edetic Acid ,Gallium Isotopes ,Aged - Abstract
OBJECTIVE: The optimal method for delineation of dominant intraprostatic lesions (DIL) for targeted radiotherapy dose escalation is unclear. This study evaluated interobserver and intermodality variability of delineations on biparametric MRI (bpMRI), consisting of T(2) weighted (T(2)W) and diffusion-weighted (DWI) sequences, and (68)Ga-PSMA-PET/CT; and compared manually delineated GTV contours with semi-automated segmentations based on quantitative thresholding of intraprostatic apparent diffusion coefficient (ADC) and standardised uptake values (SUV). METHODS: 16 patients who had bpMRI and PSMA-PET scanning performed prior to any treatment were eligible for inclusion. Four observers (two radiation oncologists, two radiologists) manually delineated the DIL on: (1) bpMRI (GTV(MRI)), (2) PSMA-PET (GTV(PSMA)) and (3) co-registered bpMRI/PSMA-PET (GTV(Fused)) in separate sittings. Interobserver, intermodality and semi-automated comparisons were evaluated against consensus Simultaneous Truth and Performance Level Estimation (STAPLE) volumes, created from the relevant manual delineations of all observers with equal weighting. Comparisons included the Dice Similarity Coefficient (DSC), mean distance to agreement (MDA) and other metrics. RESULTS: Interobserver agreement was significantly higher (p < 0.05) for GTV(PSMA) (DSC: 0.822, MDA: 1.12 mm) and GTV(Fused) (DSC: 0.787, MDA: 1.34 mm) than for GTV(MRI) (DSC: 0.705, MDA 2.44 mm). Intermodality agreement between GTV(MRI) and GTV(PSMA) was low (DSC: 0.440, MDA: 4.64 mm). Agreement between semi-automated volumes and consensus GTV was low for MRI (DSC: 0.370, MDA: 8.16 mm) and significantly higher for PSMA-PET (0.571, MDA: 4.45 mm, p < 0.05). CONCLUSION: (68)Ga-PSMA-PET appears to improve interobserver consistency of DIL localisation vs bpMRI and may be more viable for simple quantitative delineation approaches; however, more sophisticated approaches to semi-automatic delineation factoring for patient- and disease-related heterogeneity are likely required. ADVANCES IN KNOWLEDGE: This is the first study to evaluate the interobserver variability of prostate GTV delineations with co-registered bpMRI and (68)Ga-PSMA-PET.
- Published
- 2021
24. Homogeneous tumor targeting with a single dose of HER2-targeted albumin-binding domain-fused nanobody-drug conjugates results in long-lasting tumor remission in mice
- Author
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Xenaki, Katerina T, Dorrestijn, Bram, Muns, Joey A, Adamzek, Kevin, Doulkeridou, Sofia, Houthoff, HendrikJan, Oliveira, Sabrina, van Bergen En Henegouwen, Paul Mp, Sub Cell Biology, Afd Pharmaceutics, Celbiologie, Pharmaceutics, Sub Cell Biology, Afd Pharmaceutics, Celbiologie, and Pharmaceutics
- Subjects
0301 basic medicine ,Biodistribution ,Immunoconjugates ,Half-life extension ,Receptor, ErbB-2 ,media_common.quotation_subject ,Medicine (miscellaneous) ,Mice, Nude ,Antineoplastic Agents ,Toxicology and Pharmaceutics (miscellaneous) ,03 medical and health sciences ,Single-domain antibodies ,Mice ,0302 clinical medicine ,In vivo ,Albumins ,Cell Line, Tumor ,Distribution (pharmacology) ,Animals ,Humans ,Albumin-binding domain ,Aminobenzoates ,Tissue Distribution ,Internalization ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,media_common ,Pharmacology ,Mice, Inbred BALB C ,Chemistry ,fungi ,single-domain antibodies ,Albumin ,Nanobody-drug conjugates ,In vitro ,albumin-binding domain ,nanobody ,030104 developmental biology ,030220 oncology & carcinogenesis ,Cancer research ,Nanobody ,Immunohistochemistry ,half-life extension ,Female ,nanobody-drug conjugates ,Oligopeptides ,Ex vivo ,Half-Life ,Research Paper - Abstract
Background: The non-homogenous distribution of antibody-drug conjugates (ADCs) within solid tumors is a major limiting factor for their wide clinical application. Nanobodies have been shown to rapidly penetrate into xenografts, achieving more homogeneous tumor targeting. However, their rapid renal clearance can hamper their application as nanobody drug conjugates (NDCs). Here, we evaluate whether half-life extension via non-covalent interaction with albumin can benefit the efficacy of a HER2-targeted NDC. Methods: HER2-targeted nanobody 11A4 and the irrelevant nanobody R2 were genetically fused to an albumin-binding domain (ABD) at their C-terminus. Binding to both albumin and tumor cells was determined by ELISA-based assays. The internalization potential as well as the in vitro efficacy of NDCs were tested on HER2 expressing cells. Serum half-life of iodinated R2 and R2-ABD was studied in tumor-free mice. The distribution of fluorescently labelled 11A4 and 11A4-ABD was assessed in vitro in 3D spheroids. Subsequently, the in vivo distribution was evaluated by optical molecular imaging and ex vivo by tissue biodistribution and tumor immunohistochemical analysis after intravenous injection of IRDye800-conjugated nanobodies in mice bearing HER2-positive subcutaneous xenografts. Finally, efficacy studies were performed in HER2-positive NCI-N87 xenograft-bearing mice intravenously injected with a single dose (250 nmol/kg) of nanobodies conjugated to auristatin F (AF) either via a maleimide or the organic Pt(II)‑based linker, coined Lx ®. Results: 11A4-ABD was able to bind albumin and HER2 and was internalized by HER2 expressing cells, irrespective of albumin presence. Interaction with albumin did not alter its distribution through 3D spheroids. Fusion to ABD resulted in a 14.8-fold increase in the serum half-life, as illustrated with the irrelevant nanobody. Furthermore, ABD fusion prolonged the accumulation of 11A4-ABD in HER2-expressing xenografts without affecting the expected homogenous intratumoral distribution. Next to that, reduced kidney retention of ABD-fused nanobodies was observed. Finally, a single dose administration of either 11A4-ABD-maleimide-AF or 11A4-ABD-Lx-AF led to long-lasting tumor remission in HER2-positive NCI-N87 xenograft-bearing mice. Conclusion: Our results demonstrate that genetic fusion of a nanobody to ABD can significantly extend serum half-life, resulting in prolonged and homogenous tumor accumulation. Most importantly, as supported by the impressive anti-tumor efficacy observed after a single dose administration of 11A4-ABD-AF, our data reveal that monovalent internalizing ABD-fused nanobodies have potential for the development of highly effective NDCs.
- Published
- 2020
25. Effect of bovine bone collagen oligopeptides on wound healing in mice
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Di Li, Peng Liu, Teng Xu, Lin Li, Yong Li, and Jinwei Ren
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Aging ,medicine.medical_specialty ,Chemokine ,Inflammation ,Bone and Bones ,Hydroxyproline ,chemistry.chemical_compound ,Mice ,Internal medicine ,medicine ,Animals ,bovine bone ,Mice, Inbred ICR ,Wound Healing ,biology ,business.industry ,Albumin ,Interleukin ,Cell Biology ,Vascular endothelial growth factor ,Endocrinology ,chemistry ,biology.protein ,Tumor necrosis factor alpha ,Cattle ,Collagen ,medicine.symptom ,Wound healing ,business ,collagen oligopeptides ,Oligopeptides ,Research Paper - Abstract
Impaired wound healing often brings a set of problems in clinical practice. This study aimed to observe the wound healing potential of bovine bone collagen oligopeptides (BCOP) in mice. After an operation, mice in BCOP-treated groups were given intragastric administration of BCOP, while others were administered vehicle. Mice were sacrificed at different points. The wound healing condition and the tensile strength were observed, serum biochemical indexes and mRNA expression of level of related genes were measured. Compared with the normal control group, albumin (ALB), prealbumin (PA), transferrin (TRF), hydroxyproline (Hyp) levels and tension strength in the BCOP-treated groups increased significantly (p < 0.05). A pathological report showed that neutrophil granulocyte in the BCOP-treated groups decreased, while blood capillary and fibroblasts increased. The levels of serum inflammation indexes like interleukin (IL)-8, tumor necrosis factor (TNF)-α, chemokine (C-C motif) ligand 2 (CCL2) and C-reactive protein (CRP) significantly decreased in full-thickness incision model, whereas increased in full-thickness excision model (p < 0.05). Furthermore, IL-10, stromal cell-derived factor-1 alpha (SDF-1α) levels and the mRNA expression of vascular endothelial growth factor (VEGF) significantly increased in both models (p < 0.05). These results suggested that oral administration of BCOP could promote wound healing in mice.
- Published
- 2020
26. Estrogen receptor α promotes Cav1.2 ubiquitination and degradation in neuronal cells and in APP/PS1 mice
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Zhen Yan, John H. Zhang, Xiao-Juan Deng, Lu Liu, Dong Luo, Yu-Jie Lai, Xiao-Tong Hu, Bio Luo, Yan Long, Jing Tang, Yuanlin Ma, Ling He, Ming‐Jian Xiong, Jian Yang, Guojun Chen, Fei Sun, and Binglin Zhu
- Subjects
0301 basic medicine ,Aging ,Leupeptins ,Estrogen receptor ,Cav1.2 ,Amyloid beta-Protein Precursor ,Mice ,PEST sequence ,0302 clinical medicine ,Ubiquitin ,Neurons ,Estradiol ,biology ,Proto-Oncogene Proteins c-mdm2 ,Original Papers ,Cell biology ,Ubiquitin ligase ,Gene Knockdown Techniques ,Mdm2 ,Female ,Alzheimer’s disease ,Oligopeptides ,Proteasome Inhibitors ,Calcium Channels, L-Type ,Protein subunit ,Estrogen receptor α ,Mice, Transgenic ,ubiquitination ,Transfection ,K29 ,03 medical and health sciences ,Phenols ,Alzheimer Disease ,Cell Line, Tumor ,Animals ,Humans ,Cognitive Dysfunction ,Original Paper ,Estrogen Receptor alpha ,Cell Biology ,Mice, Inbred C57BL ,Disease Models, Animal ,030104 developmental biology ,Proteolysis ,biology.protein ,Pyrazoles ,030217 neurology & neurosurgery - Abstract
Cav1.2 is the pore‐forming subunit of L‐type voltage‐gated calcium channel (LTCC) that plays an important role in calcium overload and cell death in Alzheimer's disease. LTCC activity can be regulated by estrogen, a sex steroid hormone that is neuroprotective. Here, we investigated the potential mechanisms in estrogen‐mediated regulation of Cav1.2 protein. We found that in cultured primary neurons, 17β‐estradiol (E2) reduced Cav1.2 protein through estrogen receptor α (ERα). This effect was offset by a proteasomal inhibitor MG132, indicating that ubiquitin–proteasome system was involved. Consistently, the ubiquitin (UB) mutant at lysine 29 (K29R) or the K29‐deubiquitinating enzyme TRAF‐binding protein domain (TRABID) attenuated the effect of ERα on Cav1.2. We further identified that the E3 ligase Mdm2 (double minute 2 protein) and the PEST sequence in Cav1.2 protein played a role, as Mdm2 overexpression and the membrane‐permeable PEST peptides prevented ERα‐mediated Cav1.2 reduction, and Mdm2 overexpression led to the reduced Cav1.2 protein and the increased colocalization of Cav1.2 with ubiquitin in cortical neurons in vivo. In ovariectomized (OVX) APP/PS1 mice, administration of ERα agonist PPT reduced cerebral Cav1.2 protein, increased Cav1.2 ubiquitination, and improved cognitive performances. Taken together, ERα‐induced Cav1.2 degradation involved K29‐linked UB chains and the E3 ligase Mdm2, which might play a role in cognitive improvement in OVX APP/PS1 mice.
- Published
- 2019
27. Synthesis and Biological Evaluation of RGD and isoDGR-Monomethyl Auristatin Conjugates Targeting Integrin α
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André, Raposo Moreira Dias, Lizeth, Bodero, Ana, Martins, Daniela, Arosio, Silvia, Gazzola, Laura, Belvisi, Luca, Pignataro, Christian, Steinkühler, Alberto, Dal Corso, Cesare, Gennari, and Umberto, Piarulli
- Subjects
Cycloaddition Reaction ,Full Paper ,Drug Evaluation, Preclinical ,Antineoplastic Agents ,auristatins ,Full Papers ,Integrin alphaVbeta3 ,Binding, Competitive ,antitumor agents ,Cell Line, Tumor ,peptidomimetics ,drug delivery ,integrins ,Humans ,Oligopeptides - Abstract
This work reports the synthesis of a series of small‐molecule–drug conjugates containing the αVβ3‐integrin ligand cyclo[DKP‐RGD] or cyclo[DKP‐isoDGR], a lysosomally cleavable Val‐Ala (VA) linker or an “uncleavable” version devoid of this sequence, and monomethyl auristatin E (MMAE) or F (MMAF) as the cytotoxic agent. The conjugates were obtained via a straightforward synthetic scheme taking advantage of a copper‐catalyzed azide–alkyne cycloaddition as the key step. The conjugates were tested for their binding affinity for the isolated αvβ3 receptor and were shown to retain nanomolar IC50 values, in the same range as those of the free ligands. The cytotoxic activity of the conjugates was evaluated in cell viability assays with αvβ3 integrin overexpressing human glioblastoma (U87) and human melanoma (M21) cells. The conjugates possess markedly lower cytotoxic activity than the free drugs, which is consistent with inefficient integrin‐mediated internalization. In almost all cases the conjugates featuring isoDGR as integrin ligand exhibited higher potency than their RGD counterparts. In particular, the cyclo[DKP‐isoDGR]‐VA‐MMAE conjugate has low nanomolar IC50 values in cell viability assays with both cancer cell lines tested (U87: 11.50±0.13 nm; M21: 6.94±0.09 nm) and is therefore a promising candidate for in vivo experiments.
- Published
- 2019
28. RGD-expressed bacterial membrane-derived nanovesicles enhance cancer therapy
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Jin, Gao, Sihan, Wang, Xinyue, Dong, and Zhenjia, Wang
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Male ,Skin Neoplasms ,Neutrophils ,Drug Compounding ,Recombinant Fusion Proteins ,Green Fluorescent Proteins ,Tumor microenvironments ,Melanoma, Experimental ,Gene Expression ,Monocytes ,Bacterial membrane-derived nanovesicles ,Mice ,Drug Delivery Systems ,Remote loading ,Genes, Reporter ,Escherichia coli ,Tumor Microenvironment ,Animals ,Inflammation response ,Antibiotics, Antineoplastic ,Cell Membrane ,Integrin alphaVbeta3 ,Mice, Inbred C57BL ,Treatment Outcome ,Doxorubicin ,Oligopeptides ,Protein Binding ,Research Paper - Abstract
Background: A tumor microenvironment is a complicated multicellular system comprised of tumor cells, immune cells and blood vessels. Blood vessels are the barriers for drug tissue penetration. Effectively treating a cancer requires drug delivery systems to overcome biological barriers present in tumor microenvironments (TMEs). Methods: We designed a drug delivery system made of bacterial (Escherichia coli) double layer membrane-derived nanovesicles (DMVs) with the expression of RGD peptides and endogenous targeting ligands of bacteria. The physical and biological characteristics of DMVs were assessed by cryogenic transmission electron microscopy, western blotting, flow cytometry and confocal microscopy. Doxorubicin (DOX) was loaded in DMVs via a pH gradient driven drug loading method. Therapeutical effects of DOX-loaded DMVs were studied in a melanoma xenograft mouse model. Results: In vitro and in vivo experiments showed that DMVs can target neutrophils and monocytes that mediated the transport of DMVs across blood vessel barriers and they can also directly target tumor vasculature and tumor cells, resulting in enhanced delivery of therapeutics to TMEs. Furthermore, we developed a remote drug loading approach to efficiently encapsulate DOX inside DMVs, and the drug loading was 12% (w/w). In the B16-F10 melanoma mouse model, we showed that DOX-RGD-DMVs significantly inhibited the tumor growth compared to several controls. Conclusion: Our studies reveal that DMVs are a powerful tool to simultaneously target multiple cells in TMEs, thus increasing drug delivery for improved cancer therapies.
- Published
- 2020
29. A bifunctional molecule-based strategy for the development of theranostic antibody-drug conjugate
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Zhong Wu, Cao Ruiyuan, Wei Li, Lei Zhao, Fan Shiyong, Xiao Dian, Li Song, Fei Xie, Xinbo Zhou, and Lianqi Liu
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Antibody-drug conjugate ,Fluorophore ,Immunoconjugates ,theranostic ,Medicine (miscellaneous) ,Mice, Nude ,self-elimination ,Antineoplastic Agents ,010402 general chemistry ,01 natural sciences ,Cathepsin B ,law.invention ,antibody-drug conjugate ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,In vivo ,Confocal microscopy ,law ,Cell Line, Tumor ,Animals ,Humans ,Precision Medicine ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,030304 developmental biology ,0303 health sciences ,Chemistry ,Antibodies, Monoclonal ,Xenograft Model Antitumor Assays ,0104 chemical sciences ,body regions ,Monomethyl auristatin E ,7-amino-3-hydroxyethyl-coumarin ,on-off fluorescence ,Biophysics ,MCF-7 Cells ,Linker ,Oligopeptides ,Conjugate ,Research Paper - Abstract
Antibody-drug conjugates (ADCs) are being developed worldwide with the potential to revolutionize current cancer treatment strategies. Developing novel theranostic ADCs with therapeutic utility and imaging capability is an attractive and challenging subject that promises advances in the field of personalized medicine. In this work, we propose a bifunctional molecule-based strategy for the development of theranostic ADCs. Methods: We developed a theranostic ADC consisting of the anti-Her2 antibody Mil40, monomethyl auristatin E (MMAE) as the active payload, and a 7-amino-3-hydroxyethyl-coumarin (7-AHC)-based dipeptide linker, which functions as a novel bifunctional fluorescence probe that allows self-elimination cleavage in the presence of cathepsin B for payload release and fluorophore activation. The on-off fluorescence properties and the antitumor effect in vitro and in vivo were investigated. Results: A 48-fold fluorescence enhancement was observed within 1 h when the 7-AHC-based linker was exposed to cathepsin B. Cleavage upon exposure to cathepsin B allows MMAE and fluorophore intracellular release and the monitoring of MMAE distribution using confocal microscopy. Additionally, the newly developed ADC retains the advantages of traditional p-aminobenzyloxycarbonyl-containing ADCs, such as good stability (t1/2 > 7 days) and high activity in vitro (IC50 = 0.09-3.74 nM). Importantly, the theranostic ADC exhibited the equivalent antitumor efficacy to the marketed ADC T-DM1 in the classic breast cancer model. Conclusion: We suggest that the present strategy can be universally applied in all p-aminobenzyloxycarbonyl-containing ADCs. Overall, theranostic ADCs may play a role in developing new theranostic systems and promoting personalized medicine research.
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- 2020
30. Elastin-derived peptide VGVAPG decreases differentiation of mouse embryo fibroblast (3T3-L1) cells into adipocytes
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Konrad A. Szychowski, Jakub Tobiasz, Bartosz Skóra, and Jan Gmiński
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Histology ,Proteome ,Physiology ,medicine.medical_treatment ,Connective tissue ,Peptide ,macromolecular substances ,3T3 ,Diseases of the endocrine glands. Clinical endocrinology ,Hydrolysis ,Mice ,Adipokines ,3T3-L1 Cells ,medicine ,Adipocytes ,QP1-981 ,Animals ,Fibroblast ,elastin-derived peptides ,chemistry.chemical_classification ,Protease ,Adipogenesis ,integumentary system ,biology ,QH573-671 ,Chemistry ,lipid accumulation ,Embryo ,Cell Differentiation ,Cell Biology ,RC648-665 ,Lipid Metabolism ,Cell biology ,Elastin ,medicine.anatomical_structure ,VGVAPG ,cardiovascular system ,biology.protein ,Cytology ,Reactive Oxygen Species ,Oligopeptides ,Research Article ,Research Paper - Abstract
Elastin is a highly elastic protein present in connective tissue. As a result of protease activity, elastin hydrolysis occurs, and during this process, elastin-derived peptides (EDPs) are released. One of the constitutively repeating elastin and EDP building sequences is the hexapeptide VGVAPG. Therefore, the aim of our research was to define the effect of VGVAPG peptide on adipogenesis in a mouse 3T3-L1 cell line. 3T3-L1 cells were differentiated according to a previously described protocol and exposed to increasing concentrations of VGVAPG or VVGPGA peptide. The obtained results showed that VGVAPG peptide does not stimulate reactive oxygen species (ROS) production, caspase-1 activation, and 3T3-L1 cell proliferation. In the second part of the experiments, it was proved that VGVAPG peptide decreased lipid accumulation as measured by oil red O staining, which was confirmed by the profile of increased expression markers of undifferentiated preadipocytes. In our experiments, 10 nM VGVAPG added for differentiating to adipocytes increased the expression of Pref-1, serpin E1, and adiponectin as compared to rosiglitazone (PPARγ agonist)-treated group and simultaneously decreased the expression of VEGF and resistin as compared to the rosiglitazone-treated group. The obtained results show that VGVAPG peptide sustains 3T3 cells in undifferentiated state. Abbreviations DMSO: dimethyl sulphoxide; EBP: elastin-binding protein; EDPs: elastin-derived peptides; FBS: foetal bovine serum; Glb1: gene for beta-galactosidase; LDL: low-density-lipoprotein; PAI-1 (Serpin E1): plasminogen activator inhibitor-1; PBS: phosphate-buffered saline; PPARγ: peroxisome proliferator-activated receptor gamma; Pref-1: preadipocyte factor 1; ROS: reactive oxygen species; VEGF-A: vascular endothelial growth factor-A; VGVAPG: Val-Gly-Val-Ala-Pro-Gly; β-Gal: beta-galactosidase; ORO: oil red O; IBMX: 3-isobutyl-1-methylxanthine; H2DCFDA: 2ʹ,7ʹ-dichlorodihydrofluorescein diacetate; DMEM: Dulbecco’s Modified Eagle’s Medium; VVGPGA: Val-Val-Gly-Pro-Gly-Ala.
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- 2020
31. Burden of Treatment-Induced Peripheral Neuropathy in Patients with Multiple Myeloma in Sweden
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Eva Hellqvist Franck, Vishal Patel, Lucie Kutikova, Hareth Nahi, Aaron Levine, Maria Svensson, Ying Qu, Markus Hansson, Istvan Majer, and Göran Wålinder
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Adult ,Male ,medicine.medical_specialty ,Bortezomib ,chemistry.chemical_compound ,Cost of Illness ,Multiple myeloma ,Internal medicine ,Healthcare resource utilization ,Antineoplastic Combined Chemotherapy Protocols ,medicine ,Humans ,Hematologi ,Registries ,Lenalidomide ,Aged ,Retrospective Studies ,Treatment-induced peripheral neuropathy ,Aged, 80 and over ,Sweden ,Original Paper ,business.industry ,Incidence (epidemiology) ,Incidence ,Peripheral Nervous System Diseases ,Retrospective cohort study ,General Medicine ,Hematology ,Healthcare costs ,Middle Aged ,medicine.disease ,Carfilzomib ,Cancer registry ,Thalidomide ,chemistry ,Female ,business ,Multiple Myeloma ,Oligopeptides ,medicine.drug - Abstract
Introduction: Treatment-induced peripheral neuropathy (TIPN) is a complication of multiple myeloma (MM) treatment. Objective: This real-world, retrospective study used electronic medical record (EMR) data from 3 Swedish clinics to assess the occurrence and economic burden of TIPN in patients with MM. Methods: Eligible patients had an MM diagnosis in the Swedish Cancer Registry between 2006 and 2015 and initiated treatment during that period. Follow-up was until last EMR visit, death, or study end (April 2017). The current analyses included patients receiving bortezomib, lenalidomide, carfilzomib, or thalidomide at any treatment line. To discern healthcare resource utilization (HCRU) and costs associated with TIPN from other causes, patients with TIPN were matched with those without on baseline characteristics, treatment, and line of therapy. All analyses were descriptive. Results: Overall, 457 patients were included; 102 (22%) experienced TIPN. Patients experiencing TIPN during first-line treatment mostly received bortezomib-based regimens (n = 48/57 [84%]); those with TIPN during second- and third/fourth-line treatment mostly received lenalidomide/thalidomide-based regimens (19/31 [61%], 8/14 [57%], respectively). Patients with TIPN had higher HCRU/costs than those without TIPN (mean differences in hospital outpatient visits: 5.2, p = 0.0031; total costs per patient-year: EUR 17,183, p = 0.0007). Conclusions: Effective MM treatments associated with a reduced incidence of TIPN could result in decreased healthcare expenditure.
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- 2020
32. Immunohistochemical PSMA expression patterns of primary prostate cancer tissue are associated with the detection rate of biochemical recurrence with
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Daniela A, Ferraro, Jan H, Rüschoff, Urs J, Muehlematter, Benedikt, Kranzbühler, Julian, Müller, Michael, Messerli, Lars, Husmann, Thomas, Hermanns, Daniel, Eberli, Niels J, Rupp, and Irene A, Burger
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Glutamate Carboxypeptidase II ,Male ,PSMA PET ,Prostatic Neoplasms ,Androgen Antagonists ,Gallium Radioisotopes ,Middle Aged ,Prostate-Specific Antigen ,urologic and male genital diseases ,PSMA-negative prostate cancer ,Positron Emission Tomography Computed Tomography ,Antigens, Surface ,immunohistochemistry ,restaging ,Biomarkers, Tumor ,Humans ,Neoplasm Recurrence, Local ,Radiopharmaceuticals ,PSMA staining ,Oligopeptides ,Edetic Acid ,Gallium Isotopes ,Aged ,Retrospective Studies ,Research Paper - Abstract
Prostate-specific membrane antigen (PSMA) targeted PET has a high detection rate for biochemical recurrence (BCR) of prostate cancer (PCa). Nevertheless, even at high prostate-specific antigen (PSA) levels (> 3 ng/ml), a relevant number of PSMA-PET scans are negative, mainly due to PSMA-negative PCa. Our objective was to investigate whether PSMA-expression patterns of the primary tumour on immunohistochemistry (IHC) are associated with PSMA-PET detection rate of recurrent PCa. Methods: Retrospective institutional review board approved single-centre analysis of patients who had undergone 68Ga-PSMA-11-PET for BCR after radical prostatectomy (RPE) between 04/2016 and 07/2019, with tumour specimens available for PSMA-IHC. Clinical information (age, PSA-level, ongoing androgen deprivation therapy (ADT), Gleason score) and PSMA-IHC of the primary tumour were collected and their relationship to results from PSMA-PET (positive/negative) was investigated using a multiple logistic regression analysis. Results: 120 PSMA-PET scans in 74 patients were available for this analysis. Overall detection rate was 62% (74/120 scans), with a mean PSA value at scan time of 0.99 ng/ml (IQR 0.32-4.27). Of the clinical factors, only PSA-level and ADT were associated with PSMA-PET positivity. The percentage of PSMA-negative tumour area on IHC (PSMA%neg) had a significant association to PSMA-PET negativity (OR = 2.88, p < 0.001), while membranous PSMA-expression showed no association (p = 0.73). The positive predictive value of PSMA%neg ≥ 50% for a negative PSMA-PET was 85% (13/11) and for a PSMA%neg of 80% or more, 100% (9/9). Conclusions: PSMA-negative tumour area on IHC exhibited the strongest association with negative PSMA-PET scans, beside PSA-level and ADT. Even at very high PSA levels, PSMA-PET scans were negative in most of the patients with PSMA%neg ≥ 50%.
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- 2020
33. Increased recruitment of endogenous stem cells and chondrogenic differentiation by a composite scaffold containing bone marrow homing peptide for cartilage regeneration
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Jiaju Lu, Shibi Lu, Zijin Yang, Antonios G. Mikos, Jiang Peng, Yingqi Huang, Xuezhen Shen, Yu Wang, Shuhui Yang, Xiaodan Sun, Changfeng Lu, Chenhao Wang, Xiumei Wang, Quanyi Guo, Xianqi Dong, Yifan Liu, Heyong Yin, Lingyun Zhao, and Xun Sun
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0301 basic medicine ,Type II collagen ,Medicine (miscellaneous) ,02 engineering and technology ,Osteoarthritis ,Hydrogel, Polyethylene Glycol Dimethacrylate ,03 medical and health sciences ,Chondrocytes ,stem cell homing ,Cell Movement ,medicine ,Animals ,Regeneration ,self-assembling peptide hydrogel ,cartilage regeneration ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,Aggrecan ,Tissue Engineering ,Histocytochemistry ,Chemistry ,Cartilage ,Mesenchymal stem cell ,Cell Differentiation ,Mesenchymal Stem Cells ,X-Ray Microtomography ,021001 nanoscience & nanotechnology ,medicine.disease ,Chondrogenesis ,Immunohistochemistry ,Magnetic Resonance Imaging ,Cell biology ,Transplantation ,Disease Models, Animal ,Treatment Outcome ,030104 developmental biology ,medicine.anatomical_structure ,Rabbits ,Stem cell ,0210 nano-technology ,Cartilage Diseases ,Oligopeptides ,acellular cartilage matrix ,Research Paper - Abstract
Even small cartilage defects could finally degenerate to osteoarthritis if left untreated, owing to the poor self-healing ability of articular cartilage. Stem cell transplantation has been well implemented as a common approach in cartilage tissue engineering but has technical complexity and safety concerns. The stem cell homing-based technique emerged as an alternative promising therapy for cartilage repair to overcome traditional limitations. In this study, we constructed a composite hydrogel scaffold by combining an oriented acellular cartilage matrix (ACM) with a bone marrow homing peptide (BMHP)-functionalized self-assembling peptide (SAP). We hypothesized that increased recruitment of endogenous stem cells by the composite scaffold could enhance cartilage regeneration. Methods: To test our hypothesis, in vitro proliferation, attachment and chondrogenic differentiation of rabbit mesenchymal stem cells (MSCs) were tested to confirm the bioactivities of the functionalized peptide hydrogel. The composite scaffold was then implanted into full-thickness cartilage defects on rabbit knee joints for cartilage repair, in comparison with microfracture or other sample groups. Stem cell recruitment was monitored by dual labeling with CD29 and CD90 under confocal microcopy at 1 week after implantation, followed by chondrogenic differentiation examined by qRT-PCR. Repaired tissue of the cartilage defects was evaluated by histological and immunohistochemistry staining, microcomputed tomography (micro-CT) and magnetic resonance imaging (MRI) at 3 and 6 months post-surgery. Macroscopic and histological scoring was done to evaluate the optimal in vivo repair outcomes of this composite scaffold. Results: The functionalized SAP hydrogels could stimulate rabbit MSC proliferation, attachment and chondrogenic differentiation during in vitro culture. At 7 days after implantation, increased recruitment of MSCs based on CD29+ /CD90+ double-positive cells was found in vivo in the composite hydrogel scaffold, as well as upregulation of cartilage-associated genes (aggrecan, Sox9 and type II collagen). After 3 and 6 months post-surgery, the articular cartilage defect in the composite scaffold-treated group was fully covered with cartilage-like tissue with a smooth surface, which was similar to the surrounding native cartilage, according to the results of histological and immunohistochemistry staining, micro-CT and MRI analysis. Macroscopic and histological scoring confirmed that the quality of cartilage repair was significantly improved with implantation of the composite scaffold at each timepoint, in comparison with microfracture or other sample groups. Conclusion: Our findings demonstrated that the composite scaffold could enhance endogenous stem cell homing and chondrogenic differentiation and significantly improve the therapeutic outcome of chondral defects. The present study provides a promising approach for in vivo cartilage repair without cell transplantation. Optimization of this strategy may offer great potential and benefits for clinical application in the future.
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- 2018
34. Dual-functional protein for one-step production of a soluble and targeted fluorescent dye
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Yunjie Xiao, Yanyan Wang, Qian Zhang, Bin Wang, Shuxian Meng, Zefang Wang, Fengnan Sun, Yaqing Feng, Ziyu Han, and Haitao Yang
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Infrared Rays ,Hydrophobin ,Porphobilinogen ,Recombinant Fusion Proteins ,Sonication ,Mice, Nude ,Medicine (miscellaneous) ,Antineoplastic Agents ,Peptide ,02 engineering and technology ,010402 general chemistry ,01 natural sciences ,Mice ,chemistry.chemical_compound ,Nanocages ,Nanocapsules ,protein nanocage ,Cell Line, Tumor ,Neoplasms ,Animals ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,Boron ,Fluorescent Dyes ,chemistry.chemical_classification ,Microscopy, Confocal ,NIR fluorescent probes ,Imidazoles ,self-assembly ,Neoplasms, Experimental ,RGD peptide ,Flow Cytometry ,Integrin alphaVbeta3 ,021001 nanoscience & nanotechnology ,Fluorescence ,0104 chemical sciences ,Solubility ,chemistry ,Biophysics ,Female ,Self-assembly ,BODIPY ,0210 nano-technology ,Selectivity ,Oligopeptides ,Research Paper - Abstract
Low water solubility and poor selectivity are two fundamental limitations that compromise applications of near-infrared (NIR) fluorescent probes. Methods: Here, a simple strategy that can resolve these problems simultaneously was developed by using a novel hybrid protein named RGD-HFBI that is produced by fusion of hydrophobin HFBI and arginine-glycine-aspartic acid (RGD) peptide. This unique hybrid protein inherits self-assembly and targeting functions from HFBI and RGD peptide respectively. Results: Boron-dipyrromethene (BODIPY) used as a model NIR dye can be efficiently dispersed in the RGD-HFBI solution by simple mixing and sonication for 30 min. The data shows that self-assembled RGD-HFBI forms a protein nanocage by using the BODIPY as the assembly template. Cell uptake assay proves that RGD-HFBI/BODIPY can efficiently stain αvβ3 integrin-positive cancer cells. Finally, in vivo affinity tests fully demonstrate that the soluble RGD-HFBI/BODIPY complex selectively targets and labels tumor sites of tumor-bearing mice due to the high selectivity of the RGD peptide. Conclusion: Our one-step strategy using dual-functional RGD-HFBI opens a novel route to generate soluble and targeted NIR fluorescent dyes in a very simple and efficient way and may be developed as a general strategy to broaden their applications.
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- 2018
35. Antagonism of the proinflammatory and pronociceptive actions of canonical and biased agonists of protease‐activated receptor‐2
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Tina Marie Lieu, Laura E. Edgington-Mitchell, Nigel W. Bunnett, David P. Fairlie, Romke Bron, Daniel P. Poole, Peter McLean, Nicholas Barlow, E. Savage, Peishen Zhao, and Rink-Jan Lohman
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Male ,0301 basic medicine ,Proteases ,Administration, Oral ,Proinflammatory cytokine ,Rats, Sprague-Dawley ,Mice ,Structure-Activity Relationship ,03 medical and health sciences ,medicine ,Animals ,Receptor, PAR-2 ,Protease-activated receptor 2 ,Cathepsin S ,Inflammation ,Mice, Knockout ,Pharmacology ,Cathepsin ,Dose-Response Relationship, Drug ,Chemistry ,Elastase ,Nociceptors ,Research Papers ,Rats ,Cell biology ,Mice, Inbred C57BL ,030104 developmental biology ,Nociception ,Biochemistry ,Hyperalgesia ,medicine.symptom ,Oligopeptides ,Research Paper - Abstract
Background and Purpose Diverse proteases cleave protease-activated receptor-2 (PAR2) on primary sensory neurons and epithelial cells to evoke pain and inflammation. Trypsin and tryptase activate PAR2 by a canonical mechanism that entails cleavage within the extracellular N-terminus revealing a tethered ligand that activates the cleaved receptor. Cathepsin-S and elastase are biased agonists that cleave PAR2 at different sites to activate distinct signalling pathways. Although PAR2 is a therapeutic target for inflammatory and painful diseases, the divergent mechanisms of proteolytic activation complicate the development of therapeutically useful antagonists. Experimental Approach We investigated whether the PAR2 antagonist GB88 inhibits protease-evoked activation of nociceptors and protease-stimulated oedema and hyperalgesia in rodents. Key Results Intraplantar injection of trypsin, cathespsin-S or elastase stimulated mechanical and thermal hyperalgesia and oedema in mice. Oral GB88 or par2 deletion inhibited the algesic and proinflammatory actions of all three proteases, but did not affect basal responses. GB88 also prevented pronociceptive and proinflammatory effects of the PAR2-selective agonists 2-furoyl-LIGRLO-NH2 and AC264613. GB88 did not affect capsaicin-evoked hyperalgesia or inflammation. Trypsin, cathepsin-S and elastase increased [Ca2+]i in rat nociceptors, which expressed PAR2. GB88 inhibited this activation of nociceptors by all three proteases, but did not affect capsaicin-evoked activation of nociceptors or inhibit the catalytic activity of the three proteases. Conclusions and Implications GB88 inhibits the capacity of canonical and biased protease agonists of PAR2 to cause nociception and inflammation.
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- 2016
36. Development of bisphenol A-removing recombinant Escherichia coli by monomeric and dimeric surface display of bisphenol A-binding peptide
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Soon Ho Hong, Jiyeon Hong, Murali Kannan Maruthamuthu, Woo-Seok Choe, Kulandaisamy Arulsamy, Ik-Keun Yoo, and Sivachandiran Somasundaram
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0301 basic medicine ,endocrine system ,Bisphenol A ,Stereochemistry ,Bioengineering ,Peptide ,010501 environmental sciences ,medicine.disease_cause ,01 natural sciences ,03 medical and health sciences ,chemistry.chemical_compound ,Phenols ,Peptide Library ,medicine ,Escherichia coli ,Asparagine ,Benzhydryl Compounds ,Peptide library ,Peptide sequence ,0105 earth and related environmental sciences ,chemistry.chemical_classification ,urogenital system ,General Medicine ,Thermal paper ,Molecular Docking Simulation ,030104 developmental biology ,chemistry ,Bisphenol S ,Oligopeptides ,hormones, hormone substitutes, and hormone antagonists ,Biotechnology - Abstract
Peptide-displaying Escherichia coli cells were investigated for use in adsorptive removal of bisphenol A (BPA) both in Luria-Bertani medium including BPA or ATM thermal paper eluted wastewater. Two recombinant strains were constructed with monomeric and dimeric repeats of the 7-mer BPA-binding peptide (KSLENSY), respectively. Greater than threefold increased adsorption of BPA [230.4 µmol BPA per g dry cell weight (DCW)] was found in dimeric peptide-displaying cells compared to monomeric strains (63.4 µmol per g DCW) in 15 ppm BPA solution. The selective removal of BPA from a mixture of BPA analogs (bisphenol F and bisphenol S) was verified in both monomeric and dimeric peptide-displaying cells. The binding chemistry of BPA with the peptide was assumed, based on molecular docking analysis, to be the interaction of BPA with serine and asparagine residues within the 7-mer peptide sequence. The peptide-displaying cells also functioned efficiently in thermal paper eluted wastewater containing 14.5 ppm BPA.
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- 2017
37. NFAT3/c4-mediated excitotoxicity in hippocampal apoptosis during radiation-induced brain injury
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Qiuhong Fan, Ruizhe Xu, Ye Tian, Junjun Zhang, Zhao Peifeng, Liesong Chen, Yanfang Chen, and M. Xu
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Male ,0301 basic medicine ,Health, Toxicology and Mutagenesis ,Neurotoxins ,Excitotoxicity ,Hippocampus ,Apoptosis ,Hippocampal formation ,medicine.disease_cause ,Receptors, N-Methyl-D-Aspartate ,Neuroprotection ,Rats, Sprague-Dawley ,03 medical and health sciences ,0302 clinical medicine ,Bcl-2-associated X protein ,Regular Paper ,Animals ,Medicine ,Radiology, Nuclear Medicine and imaging ,calcineurin ,Radiation Injuries ,Cell Proliferation ,bcl-2-Associated X Protein ,Neurons ,NFAT3/c4 ,Radiation ,NFATC Transcription Factors ,biology ,business.industry ,Dentate gyrus ,Body Weight ,Organ Size ,NMDAR ,Calcineurin ,030104 developmental biology ,Bax ,Brain Injuries ,Dentate Gyrus ,Cancer research ,biology.protein ,Cranial Irradiation ,Prophylactic cranial irradiation ,Cognition Disorders ,business ,Oligopeptides ,030217 neurology & neurosurgery ,Signal Transduction - Abstract
Whole brain irradiation (WBI) has become an indispensible tool in the treatment of head and neck cancer, and it has greatly improved patient survival rate and total survival time. In addition, prophylactic cranial irradiation (PCI) has dramatically decreased the incidence of brain metastatic carcinoma. However, WBI may induce temporary functional deficits or even progressive, irreversible cognitive dysfunction that compromises the quality of life for survivors. Unfortunately, the exact molecular mechanisms for cognitive damage remain elusive, and no treatment or preventative measures are available for use in the clinic. In the present study, the nuclear factor of activated T cells isoform 4 (NFAT3/c4) was found to play a vital role in excitotoxic hippocampus cell apoptosis induced by radiation. Sprague–Dawley (SD) rats received 20 Gy WBI, after which we detected NFAT3/c4-mediated excitotoxicity. We found that radiation caused hippocampus excitotoxicity, resulting from overactivation of the N-methyl-D-aspartate receptor (NMDAR) and always accompanied by subsequent elevation of the intracellular calcium level and activation of calcineurin (CaN). P-NFAT3/c4 was the principal downstream target of CaN, including regulation of its nuclear translocation as well as transcriptional activities. Radiation recruited NMDAR/NFAT3/c4 activation and subsequent Bax induction in hippocampus cells. Once treated with the NFAT3/c4 inhibitor 11R-VIVIT peptide pre-irradiation, hippocampal proliferation and neuron survival (dentate gyrus cells in particular) were protected from radiation-induced injury, resulting in inhibition of the apoptosis marker Bax. Our principal aim was to illuminate the role of NFAT3/c4-mediated excitotoxicity in hippocampal apoptosis during radiation-induced brain injury. This study is the first time that radiation-induced activation of NFAT3/c4 has been recorded, and our results suggest that NFAT3/c4 may be a novel target for prevention and treatment of radiation-induced brain injury.
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- 2017
38. Spasmogenic Effects of the Proteasome Inhibitor Carfilzomib on Coronary Resistance, Vascular Tone and Reactivity
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Carol Chen-Scarabelli, Hemang Patel, Giovanni Corsetti, Riccardo Raddino, Louis D. Saravolatz, Richard Knight, Francesco S. Dioguardi, Evasio Pasini, Jagat Narula, TM Scarabelli, Mara Gavazzoni, and Gagan Sahni
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Genetics and Molecular Biology (all) ,Proteasome Endopeptidase Complex ,Spasm ,Endothelium ,lcsh:Medicine ,Vasodilation ,Proteasome inhibitors ,030204 cardiovascular system & hematology ,Pharmacology ,Biochemistry ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,0302 clinical medicine ,Nifedipine ,Multiple myeloma ,medicine.artery ,medicine ,Humans ,Vascular tone ,Carfilzomib ,Coronary resistance ,Medicine (all) ,Biochemistry, Genetics and Molecular Biology (all) ,lcsh:R5-920 ,Aorta ,business.industry ,lcsh:R ,General Medicine ,medicine.disease ,Angiotensin II ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Heart failure ,Anesthesia ,Coronary perfusion pressure ,Ventricular pressure ,Vascular Resistance ,lcsh:Medicine (General) ,business ,Oligopeptides ,Research Paper ,medicine.drug - Abstract
Background Carfilzomib (CFZ) is a new proteasome inhibitor used for the treatment of multiple myeloma. Besides heart failure, angina and myocardial ischemia occurred following administration of CFZ, which is not contraindicated in patients with recent myocardial infarction/unstable angina excluded from the safety trials. Aim of Study To test the effects of CFZ (10− 9 to 10− 7 mol/L) on vascular tone and reactivity in the isolated rabbit heart and aorta. Methods and Results CFZ administered by bolus injection to the isolated heart increased coronary perfusion pressure (CPP) at all tested concentrations and mildly raised left ventricular pressure and heart rate, only at the highest concentration. Addition of CFZ directly into the organ bath increased the basal tone of isolated aortic strips with contraction plateau reached after 10 min. This spasmogenic effect doubled following ablation of the endothelium. Pretreatment with CFZ amplified the vasospastic action exerted by KCl, noradrenaline (NA) and angiotensin II (A) on aortic strips, and impaired vasodilation following administration of nitroglycerin (NTG) and nifedipine (NFP) on the contraction plateau induced by KCl, NA and A. Aortic strips pretreated with CFZ exhibited impaired relaxation, as compared to untreated strips, following administration of acetylcholine (Ach), an endothelium-dependent vasodilating agent, on the plateau of NA contraction (p, Highlights • In the isolated aorta, carfilzomib increased basal tone and vasospastic action of KCl, noradrenaline and angiotensin II. • In the isolated aorta, carfilzomib impaired the anti-spasmogenic activity of nitroglycerin, nifedipine and acetylcholine. • In the isolated heart, carfilzomib increased coronary perfusion pressure, and mildly left ventricular pressure and heart rate. Carfilzomib is a new chemotherapeutic agent used for the treatment of multiple myeloma. Our study shows that carfilzomib increases coronary perfusion pressure, resting vasoconstricting tone, and the spasmogenic effect of noradrenaline and angiotensin II, while it curbs the vasodilatory action of nitroglycerine and nifedipine. Our findings are relevant to human health as they warrant caution in the use of carfilzomib in elderly patients with cardiovascular risk factors and, even more importantly, in those with preexisting heart conditions, who are also eligible to receive carfilzomib, even though they were excluded from the safety trials, based on which carfilzomib use was approved.
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- 2017
39. Glutaminase inhibitor CB-839 synergizes with carfilzomib in resistant multiple myeloma cells
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Nathan G. Dolloff, Brittany Smith, Amit Kumar Mitra, Yefim Manevich, Dominik Dytfeld, Andrzej Jakubowiak, Mieczysław Komarnicki, Tomasz Szczepaniak, Ravyn M. Thompson, Reeder M. Robinson, Brian G Van Ness, Leticia Reyes, Anna Przybylowicz-Chalecka, and Magdalena Luczak
- Subjects
0301 basic medicine ,Cell Survival ,Cell Respiration ,Benzeneacetamides ,Antineoplastic Agents ,Apoptosis ,03 medical and health sciences ,chemistry.chemical_compound ,Cell Line, Tumor ,Antineoplastic Combined Chemotherapy Protocols ,Thiadiazoles ,medicine ,Humans ,Cytotoxic T cell ,Multiple myeloma ,Aged ,carfilzomib ,proteasome inhibitor ,business.industry ,Bortezomib ,Glutaminase ,Drug Synergism ,glutaminase ,Endoplasmic Reticulum Stress ,medicine.disease ,Carfilzomib ,Mitochondria ,3. Good health ,multiple myeloma ,030104 developmental biology ,Oncology ,chemistry ,Drug Resistance, Neoplasm ,Immunology ,Proteasome inhibitor ,Cancer research ,Energy Metabolism ,business ,Glutaminase Inhibitor CB-839 ,Oligopeptides ,Proteasome Inhibitors ,Biomarkers ,Research Paper ,medicine.drug - Abstract
// Ravyn M. Thompson 1 , Dominik Dytfeld 2 , Leticia Reyes 1 , Reeder M. Robinson 1 , Brittany Smith 1 , Yefim Manevich 1 , Andrzej Jakubowiak 3 , Mieczyslaw Komarnicki 2 , Anna Przybylowicz-Chalecka 2 , Tomasz Szczepaniak 2 , Amit K. Mitra 5 , Brian G. Van Ness 5 , Magdalena Luczak 4 , Nathan G. Dolloff 1, * 1 Department of Cell and Molecular Pharmacology & Experimental Therapeutics, Medical University of South Carolina, Charleston, SC, USA 2 Karol Marcinkowski University of Medical Sciences, Poznan, Poland 3 University of Chicago, Chicago, IL, USA 4 Institute of Bioorganic Chemistry, Polish Academy of Sciences, Poznan, Poland 5 University of Minnesota, Minneapolis, MN, USA * These authors contributed equally to this work Correspondence to: Nathan G. Dolloff, email: dolloffn@musc.edu Keywords: multiple myeloma, proteasome inhibitor, glutaminase, carfilzomib Received: December 08, 2016 Accepted: March 01, 2017 Published: March 16, 2017 ABSTRACT Curative responses in the treatment of multiple myeloma (MM) are limited by the emergence of therapeutic resistance. To address this problem, we set out to identify druggable mechanisms that convey resistance to proteasome inhibitors (PIs; e.g., bortezomib), which are cornerstone agents in the treatment of MM. In isogenic pairs of PI sensitive and resistant cells, we observed stark differences in cellular bioenergetics between the divergent phenotypes. PI resistant cells exhibited increased mitochondrial respiration driven by glutamine as the principle fuel source. To target glutamine-induced respiration in PI resistant cells, we utilized the glutaminase-1 inhibitor, CB-839. CB-839 inhibited mitochondrial respiration and was more cytotoxic in PI resistant cells as a single agent. Furthermore, we found that CB-839 synergistically enhanced the activity of multiple PIs with the most dramatic synergy being observed with carfilzomib (Crflz), which was confirmed in a panel of genetically diverse PI sensitive and resistant MM cells. Mechanistically, CB-839 enhanced Crflz-induced ER stress and apoptosis, characterized by a robust induction of ATF4 and CHOP and the activation of caspases. Our findings suggest that the acquisition of PI resistance involves adaptations in cellular bioenergetics, supporting the combination of CB-839 with Crflz for the treatment of refractory MM.
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- 2017
40. Targeting PEPT1: a novel strategy to improve the antitumor efficacy of doxorubicin in human hepatocellular carcinoma therapy
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Qingyu Zhang, Jia Zhao, Xu Jian, Xiang Wu, Tao Wang, Zhi Yao, Yanxia Gong, and Xi Liu
- Subjects
0301 basic medicine ,Carcinoma, Hepatocellular ,Side effect ,Tumor target ,Mice, Nude ,Pharmacology ,doxorubicin ,Peptide Transporter 1 ,hepatocellular carcinoma (HCC) ,Mice ,03 medical and health sciences ,0302 clinical medicine ,In vivo ,Cell Line, Tumor ,Animals ,Humans ,Medicine ,Doxorubicin ,Target therapy ,Oligopeptide ,Symporters ,target therapy ,business.industry ,Liver Neoplasms ,toxicity ,Biological Transport ,Drug Synergism ,SUPERFAMILY ,Hep G2 Cells ,medicine.disease ,Xenograft Model Antitumor Assays ,digestive system diseases ,proton coupled oligopeptide transporter 1 ,030104 developmental biology ,Oncology ,030220 oncology & carcinogenesis ,Hepatocellular carcinoma ,Cancer research ,Caco-2 Cells ,business ,Oligopeptides ,Research Paper ,medicine.drug - Abstract
// Yanxia Gong 1, 2 , Xiang Wu 3 , Tao Wang 1 , Jia Zhao 3 , Xi Liu 1 , Zhi Yao 4 , Qingyu Zhang 1 and Xu Jian 3 1 Department of Gastroenterology, Tianjin Medical University General Hospital, Tianjin, 300052, China 2 Department of Gastroenterology, Tianjin Nankai Hospital, Tianjin, 300100, China 3 Central Laboratory, Tianjin Medical University General Hospital, Tianjin, 300052, China 4 Department of Immunology, School of Basic Medical Science, Tianjin Medical University, Tianjin, 300070, China Correspondence to: Xu Jian, email: jianxu516@hotmail.com Qingyu Zhang, email: zhangqy@tijmu.edu.cn Keywords: proton coupled oligopeptide transporter 1, doxorubicin, hepatocellular carcinoma (HCC), target therapy, toxicity Received: September 02, 2016 Accepted: April 02, 2017 Published: April 15, 2017 ABSTRACT Proton coupled oligopeptide transporter 1 (PEPT1) is a member of the peptide transporter superfamily and plays important role in the absorption of oligopeptide and peptidomimetic drugs. Our previous research verified that PEPT1 expressed specifically in human Hepatocellular carcinoma (HCC) tissue and cell lines and showed potential transport activity to be a new candidate of the tumor therapeutic target. In this study, we aim to explore the feasibility of a novel tumor target therapeutic strategy: Targeting PEPT1 to improve the antitumor efficacy of Doxorubicin in human HCC therapy. First, Doxorubicin was conjugated with Glycylglycylglycine (Gly-Gly-Gly) − a tripeptide which was known as the substrate of PEPT1 and characterized by HPLC and MS successfully. Doxorubicin-tripeptide conjugate was then observed to clarify the target delivery by PEPT1 and the antitumor effect on human hepatocarcinoma in vivo and in vitro . Furthermore, the improvement of the toxic and side effect of Doxorubicin after conjugation was also evaluated by some biochemical tests. Our results reveal that targeting PEPT1 may contribute to the efficient delivery of Doxorubicin to hepatocarcinoma cells and the reduction of drug toxicity. PEPT1 has the prospect to be a novel target of HCC therapy.
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- 2017
41. Targeting of apoptotic pathways by SMAC or BH3 mimetics distinctly sensitizes paclitaxel-resistant triple negative breast cancer cells
- Author
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Anna-Katharina Müller, Sima Lev, Guohong Hu, Effrosini Panayotopoulou, Hauke Busch, and Melanie Börries
- Subjects
SMAC mimetics ,0301 basic medicine ,Indoles ,Paclitaxel ,medicine.medical_treatment ,Apoptosis ,Triple Negative Breast Neoplasms ,Pharmacology ,Piperazines ,Targeted therapy ,resistance ,Mitochondrial Proteins ,Nitrophenols ,03 medical and health sciences ,chemistry.chemical_compound ,Downregulation and upregulation ,Biomimetic Materials ,Cell Line, Tumor ,Proto-Oncogene Proteins ,medicine ,Humans ,Molecular Targeted Therapy ,high-throughput screen ,Triple-negative breast cancer ,Sulfonamides ,Chemotherapy ,Aniline Compounds ,Taxane ,business.industry ,Biphenyl Compounds ,Intracellular Signaling Peptides and Proteins ,Dipeptides ,Molecular medicine ,Peptide Fragments ,030104 developmental biology ,Oncology ,chemistry ,Drug Resistance, Neoplasm ,triple negative breast cancer ,FOXM1 ,Cancer research ,Female ,Apoptosis Regulatory Proteins ,business ,Oligopeptides ,Research Paper ,Signal Transduction - Abstract
// Effrosini G. Panayotopoulou 1 , Anna-Katharina Muller 1 , Melanie Borries 2 , Hauke Busch 2 , Guohong Hu 3 , Sima Lev 1 1 Molecular Cell Biology Department, Weizmann Institute of Science, Rehovot 76100, Israel 2 Institute of Molecular Medicine and Cell Research (IMMZ), Albert Ludwigs-University, 79104 Freiburg, Germany 3 Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China Correspondence to: Sima Lev, email: sima.lev@weizmann.ac.il Keywords: triple negative breast cancer, paclitaxel, resistance, SMAC mimetics, high-throughput screen Received: September 13, 2016 Accepted: January 24, 2017 Published: February 06, 2017 ABSTRACT Standard chemotherapy is the only systemic treatment for triple-negative breast cancer (TNBC), and despite the good initial response, resistance remains a major therapeutic obstacle. Here, we employed a High-Throughput Screen to identify targeted therapies that overcome chemoresistance in TNBC. We applied short-term paclitaxel treatment and screened 320 small-molecule inhibitors of known targets to identify drugs that preferentially and efficiently target paclitaxel-treated TNBC cells. Among these compounds the SMAC mimetics (BV6, Birinapant) and BH3-mimetics (ABT-737/263) were recognized as potent targeted therapy for multiple paclitaxel-residual TNBC cell lines. However, acquired paclitaxel resistance through repeated paclitaxel pulses result in desensitization to BV6, but not to ABT-263, suggesting that short- and long-term paclitaxel resistance are mediated by distinct mechanisms. Gene expression profiling of paclitaxel-residual, -resistant and naive MDA-MB-231 cells demonstrated that paclitaxel-residual, as opposed to -resistant cells, were characterized by an apoptotic signature, with downregulation of anti-apoptotic genes ( BCL2, BIRC5 ), induction of apoptosis inducers ( IL24, PDCD4 ), and enrichment of TNFα/NF-κB pathway, including upregulation of TNFSF15 , coupled with cell-cycle arrest. BIRC5 and FOXM1 downregulation and IL24 induction was also evident in breast cancer patient datasets following taxane treatment. Exposure of naive or paclitaxel-resistant cells to supernatants of paclitaxel-residual cells sensitized them to BV6, and treatment with TNFα enhanced BV6 potency, suggesting that sensitization to BV6 is mediated, at least partially, by secreted factor(s). Our results suggest that administration of SMAC or BH3 mimetics following short-term paclitaxel treatment could be an effective therapeutic strategy for TNBC, while only BH3-mimetics could effectively overcome long-term paclitaxel resistance.
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- 2017
42. The pentapeptide Gly-Thr-Gly-Lys-Thr confers sensitivity to anti-cancer drugs by inhibition of CAGE binding to GSK3β and decreasing the expression of cyclinD1
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Hansoo Lee, Doyong Jeon, Youngmi Kim, Yun Sil Lee, Hyun-A Kim, Dooil Jeoung, Deokbum Park, Jongseon Choe, and Young Myeong Kim
- Subjects
Male ,0301 basic medicine ,Protein domain ,Mice, Nude ,Antineoplastic Agents ,Peptide ,Pentapeptide repeat ,DEAD-box RNA Helicases ,cyclinD1 ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Protein Domains ,Antigen ,Cell Movement ,Cell Line, Tumor ,Animals ,Humans ,Medicine ,Cyclin D1 ,Amino Acid Sequence ,Promoter Regions, Genetic ,Peptide sequence ,chemistry.chemical_classification ,Mice, Inbred BALB C ,Oligopeptide ,Glycogen Synthase Kinase 3 beta ,business.industry ,anti-cancer drug-resistance ,GSK3β ,Molecular biology ,Amino acid ,030104 developmental biology ,Oncology ,chemistry ,Drug Resistance, Neoplasm ,CAGE ,030220 oncology & carcinogenesis ,Cancer cell ,Immunology ,peptides ,Female ,business ,Oligopeptides ,Research Paper - Abstract
// Youngmi Kim 1, * , Hyuna Kim 1, * , Deokbum Park 1 , Hansoo Lee 2 , Yun Sil Lee 3 , Jongseon Choe 4 , Young Myeong Kim 4 , Doyong Jeon 5 , Dooil Jeoung 1 1 Department of Biochemistry, College of Natural Sciences, Kangwon National University, Chunchon 24341, Korea 2 Department of Biological Sciences, College of Natural Sciences, Kangwon National University, Chunchon 24341, Korea 3 College of Pharmacy, Ewha Womans University, Seoul 03760, Korea 4 Graduate School of Medicine, Kangwon National University, Chunchon 24341, Korea 5 L-Base Company, Seoul 01062, Korea * These authors contributed equally to this work Correspondence to: Dooil Jeoung, email: jeoungd@kangwon.ac.kr Keywords: anti-cancer drug-resistance, CAGE, cyclinD1, GSK3β, peptides Received: February 12, 2016 Accepted: January 03, 2017 Published: January 13, 2017 ABSTRACT We previously reported the role of cancer/testis antigen CAGE in the response to anti-cancer drugs. CAGE increased the expression of cyclinD1, and pGSK3β Ser9 , an inactive GSK3β, while decreasing the expression of phospho-cyclinD1Thr 286 . CAGE showed binding to GSK3β and the domain of CAGE (amino acids 231–300) necessary for binding to GSK3β and for the expression regulation of cyclinD1 was determined. 269 GTGKT 273 peptide, corresponding to the DEAD box helicase domain of CAGE, decreased the expression of cyclinD1 and pGSK3β Ser9 while increasing the expression of phospho-cyclinD1 Thr286 . GTGKT peptide showed the binding to CAGE and prevented CAGE from binding to GSK3β. GTGKT peptide changed the localization of CAGE and inhibited the binding of CAGE to the promoter sequences of cyclin D1. GTGKT peptide enhanced the apoptotic effects of anti-cancer drugs and decreased the migration, invasion, angiogenic, tumorigenic and metastatic potential of anti-cancer drug-resistant cancer cells. We found that Lys 272 of GTGKT peptide was necessary for conferring anti-cancer activity. Peptides corresponding to the DEAD box helicase domain of CAGE, such as AQTGTGKT, QTGTGKT and TGTGKT, also showed anti-cancer activity by preventing CAGE from binding to GSK3β. GTGKT peptide showed ex vivo tumor homing potential. Thus, peptides corresponding to the DEAD box helicase domain of CAGE can be developed as anti-cancer drugs in cancer patients expressing CAGE.
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- 2017
43. A Nanosystem of Amphiphilic Oligopeptide-Drug Conjugate Actualizing Both αvβ3 Targeting and Reduction-Triggered Release for Maytansinoid
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Qiang Zhang, Suxin Li, Yanqin Liang, Demin Zhou, Wenbing Dai, Yiguang Wang, Xiaoyou Wang, Bo He, Bing He, Hua Zhang, and Xueqing Wang
- Subjects
maytansinoid DM1 ,Cell Survival ,antitumor therapy ,Melanoma, Experimental ,Medicine (miscellaneous) ,Antineoplastic Agents ,02 engineering and technology ,Pharmacology ,010402 general chemistry ,Maytansinoid ,01 natural sciences ,αvβ3-targeted nanoparticles ,Surface-Active Agents ,chemistry.chemical_compound ,In vivo ,Human Umbilical Vein Endothelial Cells ,endocytosis ,Animals ,Humans ,Maytansine ,Particle Size ,Cytotoxicity ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,Amphiphilic peptide-drug conjugate ,Chromatography, High Pressure Liquid ,Oligopeptide ,Body Weight ,Cell Cycle ,Prodrug ,Integrin alphaVbeta3 ,021001 nanoscience & nanotechnology ,Small molecule ,In vitro ,0104 chemical sciences ,Mice, Inbred C57BL ,Drug Liberation ,chemistry ,Hydrodynamics ,MCF-7 Cells ,Nanoparticles ,0210 nano-technology ,reduction-triggered drug release ,Oligopeptides ,Linker ,Research Paper - Abstract
To design a prodrug-based self-assembling nanosystem with both ligand targeting and stimuli-responsive features, and elucidate the superiority of each targeting strategy and the synergistic effect between them, we synthesized four small molecule amphiphilic peptide-drug conjugates (APDCs) using maytansinoid (DM1) as a cytotoxic agent, cRGDfK as a homing peptide, and disulfide (SS) or thioether (SMCC) as linker. Owing to their amphiphilicity, the APDCs could self-assemble into nanoparticles (APDC@NPs) which were evaluated in vitro in three different cell lines and in vivo in tumor-bearing C57BL/6 mice. The RSSD@NPs showed the strongest interaction with αvβ3 integrin, highest cell uptake and intracellular free drug level, and best antitumor efficacy in vitro and in vivo, while it shared the same goodness with other test nanosystems in terms of high drug loading, EPR effect and free of potentially toxic polymers. Especially, the in vivo efficacy of RSSD@NPs was 2 fold of free DM1 which is too cytotoxic to be a drug, while the active targeted APDC@NPs demonstrated acceptable system, tissue and blood compatibility. In αvβ3-positive cells or tumors, the RGD targeting contributed much more than disulfide in anticancer effect. The maximum synergism of the two strategies reached to 22 fold in vitro and 3 fold in vivo. Generally, the active targeting, prodrug and nanosystem could significantly decrease the toxicity of free DM1 and improve its therapy outcome via combining active targeting, prodrug and nanopreparation, especially the dual targeting strategies and their synergism.
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- 2017
44. SS31 Ameliorates age-related activation of NF-κB signaling in senile mice model, SAMP8
- Author
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Tian-Tian Huo, Yue Huang, Jian-Hua Wang, Ping Fan, Qian Jia, Zhi-Hua Hao, Rong-Fang Feng, and Mei-Rong Wang
- Subjects
Male ,0301 basic medicine ,Gerontology ,Senescence ,Aging ,medicine.medical_specialty ,Inflammation ,medicine.disease_cause ,Antioxidants ,NF-κB ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,Research Paper: Gerotarget (Focus on Aging) ,0302 clinical medicine ,Internal medicine ,Gene expression ,medicine ,Animals ,Hippocampus (mythology) ,Cognitive Dysfunction ,SAMP8 ,Gerotarget ,business.industry ,NF-kappa B ,Blot ,Disease Models, Animal ,Oxidative Stress ,030104 developmental biology ,Endocrinology ,Oncology ,chemistry ,inflammation ,medicine.symptom ,Signal transduction ,Reactive Oxygen Species ,business ,Oligopeptides ,030217 neurology & neurosurgery ,Oxidative stress ,Signal Transduction ,SS31 - Abstract
// Zhi-Hua Hao 1,2 , Yue Huang 3 , Mei-Rong Wang 1,2 , Tian-Tian Huo 1 , Qian Jia 2 , Rong-Fang Feng 1 , Ping Fan 2 and Jian-Hua Wang 1 1 Department of Neurology, Hebei General Hospital, Shijiazhuang, Hebei, China 2 Graduate School,Hebei Medical University, Shijiazhuang, Hebei, China 3 School of Medical Sciences, Faculty of Medicine, UNSW Australia, Sydney, Australia Correspondence to: Jian-Hua Wang, email: // Keywords : aging, SAMP8, inflammation, SS31, NF-κB, Gerotarget Received : March 31, 2016 Accepted : December 12, 2016 Published : December 21, 2016 Abstract Aging has been attributed to oxidative stress and inflammatory response, in which NF-κB and Nrf2-ARE signaling pathways play significant roles. Senescence accelerated mouse prone 8 (SAMP8) is generally used an animal model for aging studies. Here, we investigated the NF-κB and Nrf2-ARE signaling pathways in SAMP8 brains at different ages and their responses to SS31 peptide treatment. Thirty six SAMP8 mice were separated into aging groups and SS31-treatment groups. The hippocampus from each mouse was dissected for RNA and protein extraction. Cytokines and ROS levels were measured using ELISA and standardised method. Gene expressions of NF-κB, Nrf2 and HO-1 were measured by RT-qPCR. Total protein amount of NF-κB and HO-1, as well as the concentrations of nuclear and cytoplasmic Nrf2 were measured using Western blots. Our data showed that aging could activate both NF-κB and Nrf2-ARE signaling pathways, which could be suppressed and activated by SS31 treatment respectively. Regression analysis revealed that NF-κB gene expression was the most important parameter predicting aging process and SS31 treatment effects in SAMP8. Our findings suggested that SS31 treatment may modulate the inflammatory and oxidative stress status of the aged brains and exert protective effects during brain aging.
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- 2016
45. Precise integrin-targeting near-infrared imaging-guided surgical method increases surgical qualification of peritoneal carcinomatosis from gastric cancer in mice
- Author
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Jinzuo Ye, Jianxin Cui, Jie Tian, Haidong Cheng, Huiping Huo, Sha Rengaowa, Lin Chen, Caoting Zeng, Wenting Shang, Jiang Shixin, Yamin Mao, and Chongwei Chi
- Subjects
Indocyanine Green ,0301 basic medicine ,medicine.medical_specialty ,Time Factors ,integrin ,Mice, Nude ,fluorescence molecular imaging ,03 medical and health sciences ,Peritoneal Neoplasm ,0302 clinical medicine ,Quality of life ,Predictive Value of Tests ,Stomach Neoplasms ,Cytoreduction Surgical Procedures ,Biomarkers, Tumor ,medicine ,Animals ,Humans ,Peritoneal Neoplasms ,Fluorescent Dyes ,Mice, Inbred BALB C ,Spectroscopy, Near-Infrared ,business.industry ,gastric cancer ,General surgery ,Metastasectomy ,peritoneal carcinomatosis ,Cancer ,Integrin alphaVbeta3 ,medicine.disease ,Xenograft Model Antitumor Assays ,Molecular Imaging ,Surgery ,Peritoneal carcinomatosis ,030104 developmental biology ,Surgery, Computer-Assisted ,Oncology ,030220 oncology & carcinogenesis ,Predictive value of tests ,Luminescent Measurements ,MCF-7 Cells ,Female ,Hyperthermic intraperitoneal chemotherapy ,business ,Oligopeptides ,Research Paper - Abstract
// Haidong Cheng 1, 2, 3, * , Chongwei Chi 3, * , Wenting Shang 3, * , Sha Rengaowa 4 , Jianxin Cui 1 , Jinzuo Ye 3 , Shixin Jiang 3 , Yamin Mao 3 , Caoting Zeng 5 , Huiping Huo 6 , Lin Chen 1 , Jie Tian 3 1 Department of General Surgery, The Chinese PLA General Hospital, Beijing, 100853, China 2 Department of General Surgery, The First Affiliated Hospital of Inner Mongolia Medical University, Hohhot, 010059, China 3 Key Laboratory of Molecular Imaging of the Chinese Academy of Sciences, Institute of Automation, Chinese Academy of Sciences, Beijing, 100190, China 4 Department of Basic Medical Science, Inner Mongolia Medical University, Hohhot, 010059, China 5 Department of Hepatobiliary Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou, 510280, China 6 Department of Ultrasound, General Hospital of the People’s Liberation Army, Beijing, 100853, China * These authors contributed equally to this work Correspondence to: Jie Tian, email: tian@ieee.org , jie.tian@ia.ac.cn Lin Chen, email: chenlin301@yeah.net Keywords: gastric cancer, fluorescence molecular imaging, peritoneal carcinomatosis, integrin Received: July 18, 2016 Accepted: December 12, 2016 Published: December 21, 2016 ABSTRACT Peritoneal carcinomatosis from gastric cancer represents a common recurrent gastric cancer that seriously affects the survival, prognosis, and quality of life of patients at its advanced stage. In recent years, complete cytoreduction surgery in combination with hyperthermic intraperitoneal chemotherapy has been demonstrated to improve the survival and prognosis of patients with malignant tumors including peritoneal carcinomatosis from gastric cancer. Establishing viable methods of accurately assessing the tumor burden in patients with peritoneal carcinoma and correctly selecting suitable patients in order to improve cytoreduction surgical outcomes and reduce the risk of postoperative complications has become a challenge in the field of peritoneal carcinoma research. Here, we investigated peritoneal carcinomatosis from gastric cancer in a mouse model by using our self-developed surgical navigation system that combines optical molecular imaging with an integrin-targeting Arg-Gly-Asp-indocyanine green (RGD-ICG) molecular probe. The results showed that our diagnostic method could achieve a sensitivity and specificity of up to 93.93% and 100%, respectively, with a diagnostic index (DI) of 193.93% and diagnostic accuracy rate of 93.93%.Furthermore, the minimum tumor diameter measured during the surgery was 1.8 mm and the operative time was shortened by 3.26-fold when compared with the conventionally-treated control group. Therefore, our surgical navigation system that combines optical molecular imaging with an RGD-ICG molecular probe, could improve the diagnostic accuracy rate for peritoneal carcinomatosis from gastric cancer, shorten the operative time, and improve the quality of the cytoreduction surgery for peritoneal carcinomatosis from gastric cancer, thus providing a solid foundation for its future clinical development and application.
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- 2016
46. Synthesis of a cell penetrating peptide modified superparamagnetic iron oxide and MRI detection of bladder cancer
- Author
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Chen Ding, Jinhai Fan, Kaijie Wu, Lei Wang, Xinyang Wang, Li Liu, Weiyi Wang, Rong Wang, and Zhenfeng Guan
- Subjects
0301 basic medicine ,Pathology ,medicine.medical_specialty ,Neoplasm, Residual ,Urinary system ,Contrast Media ,Cell-Penetrating Peptides ,02 engineering and technology ,targeted diagnosis ,Malignancy ,Ferric Compounds ,03 medical and health sciences ,chemistry.chemical_compound ,Cell Line, Tumor ,Humans ,Medicine ,Magnetite Nanoparticles ,Fluorescein isothiocyanate ,Urine cytology ,Bladder cancer ,Dose-Response Relationship, Drug ,medicine.diagnostic_test ,business.industry ,Magnetic resonance imaging ,021001 nanoscience & nanotechnology ,medicine.disease ,Magnetic Resonance Imaging ,Minimal residual disease ,030104 developmental biology ,Urinary Bladder Neoplasms ,cell penetrating peptide ,Oncology ,chemistry ,Cell-penetrating peptide ,bladder cancer ,superparamagnetic iron oxide ,Lysosomes ,0210 nano-technology ,business ,Oligopeptides ,Fluorescein-5-isothiocyanate ,Research Paper - Abstract
// Chen Ding 1, 2 , Kaijie Wu 1 , Weiyi Wang 1 , Zhenfeng Guan 1 , Lei wang 3 , Xinyang Wang 4 , Rong Wang 5 , Li Liu 6 , Jinhai Fan 1, 4 1 Department of Urology, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China 2 Department of Urology, Xiangyang Central Hospital, Hubei University of Arts and Science, Hubei Province, China 3 Department of Thoracic Surgery, Tangdu Hospital, The Forth Military Medical University, Xi’an, China 4 Oncology Research Lab, Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education, Xi’an, China 5 Department of Radiology, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China 6 Department of Radiology, The University of Texas Southwestern Medical Center, Dallas, TX, USA Correspondence to: Li Liu, email: li.liu@utsouthwestern.edu Jinhai Fan, email: jinhaif029@126.com Keywords: bladder cancer, superparamagnetic iron oxide, cell penetrating peptide, magnetic resonance imaging, targeted diagnosis Received: May 18, 2016 Accepted: November 01, 2016 Published: November 24, 2016 ABSTRACT Bladder cancer is the most common malignancy of the urinary tract for which the accurate measurement of minimal residual disease is critical to treatment and determining prognosis. Although cystoscope examination and voided urine cytology remain the current standard of care for detecting residual disease, these approaches are limited by mechanical trauma and lack sensitivity. To develop a new accurate noninvasive method, we developed a novel contrast agent where the surface of superparamagnetic iron oxide (SPIO) nanoparticles is functionalized with a bladder cancer-specific fluorescein isothiocyanate (FITC) labeled cell penetrating peptide (CPP)-polyarginine peptides (R11) for active targeting and imaging. The stable nanoparticles have an average hydrodynamic diameter of 51 nm, surface charge of -21 mV and MRI r 2 relaxivity 135 mM -1 s -1 . In vitro cell studies demonstrated that the R11-conjugated SPIO (SPIO-R11) nanoparticles were taken up by bladder cancer cells (T24) in a dose-dependent manner, which was higher than unconjugated SPIO. TEM showed that SPIO-R11 was mainly concentrated on cell vesicle and lysosome, not in cell nucleus, and no obvious damage was seen on cell ultrastructure. Moreover, uptake of the nanoparticles showed significantly more SPIO-R11 accumulation in bladder cancer cells than in immortalized bladder epithelial cells unlike control SPIO. Further, SPIO-R11 was compatible with immortalized bladder epithelial cells at all tested concentrations up to 200 μg/mL after 72 h incubation. Moreover, SPIO-R11 decreased the magnetic resonance T 2 relaxation time by 73% in tumors cells in vitro compared to 12% with SPIO. These results indicate great potential of SPIO-R11 as contrast agent to target bladder cancer for diagnostic and therapeutic applications.
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- 2016
47. Combining a GSI and BCL-2 inhibitor to overcome melanoma's resistance to current treatments
- Author
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Josianna V. Schwan, Yuchun Luo, Nabanita Mukherjee, Yan Lu, Yiqun G. Shellman, Allison J. Applegate, Katie A. Partyka, Adam R. Almeida, Steven E. Robinson, David A. Norris, William A. Robinson, Madison A. Rogers, Mayumi Fujita, Carol M. Amato, and K. Lambert
- Subjects
0301 basic medicine ,Neuroblastoma RAS viral oncogene homolog ,medicine.medical_treatment ,Aldehyde dehydrogenase ,Apoptosis ,Piperazines ,GTP Phosphohydrolases ,Nitrophenols ,Mice ,0302 clinical medicine ,Cell Self Renewal ,Melanoma ,Sulfonamides ,Neurofibromin 1 ,biology ,Drug Synergism ,melanoma stem cells ,3. Good health ,melanoma initiating cells ,Proto-Oncogene Proteins c-bcl-2 ,Oncology ,030220 oncology & carcinogenesis ,Neoplastic Stem Cells ,Drug Therapy, Combination ,Female ,Oligopeptides ,Research Paper ,Proto-Oncogene Proteins B-raf ,Programmed cell death ,Mice, Nude ,GSI-I ,03 medical and health sciences ,Cell Line, Tumor ,medicine ,Animals ,Humans ,Viability assay ,ABT-737 ,business.industry ,Biphenyl Compounds ,Membrane Proteins ,Immunotherapy ,medicine.disease ,Xenograft Model Antitumor Assays ,Bcl-2 Inhibitor ,030104 developmental biology ,Drug Resistance, Neoplasm ,BRAF-WT melanoma ,Mutation ,Immunology ,Cancer research ,biology.protein ,Amyloid Precursor Protein Secretases ,Neoplasm Recurrence, Local ,business - Abstract
Major limitations of current melanoma treatments are for instances of relapse and the lack of therapeutic options for BRAF wild-type patients who do not respond to immunotherapy. Many studies therefore focus on killing resistant subpopulations, such as Melanoma Initiating Cells (MICs) to prevent relapse. Here we examined whether combining a GSI (γ-Secretase Inhibitor) with ABT-737 (a small molecule BCL-2/BCL-XL/BCL-W inhibitor) can kill both the non-MICs (bulk of melanoma) and MICs. To address the limitations of melanoma therapies, we included multiple tumor samples of patients relapsed from current treatments, with a diverse genetic background (with or without the common BRAF, NRAS or NF1 mutations) in these studies. Excitingly, the combination treatment reduced cell viability and induced apoptosis of the non-MICs; disrupted primary spheres, decreased the ALDH+ cells, and inhibited the self-renewability of the MICs in multiple melanoma cell lines and relapsed patient samples. Using a low-cell-number mouse xenograft model, we demonstrated that the combination significantly reduced the tumor initiating ability of MIC-enriched cultures from relapsed patient samples. Mechanistic studies also indicate that cell death is NOXA-dependent. In summary, this combination may be a promising strategy to address treatment relapse and for triple wild-type patients who do not respond to immunotherapy.
- Published
- 2016
48. Immune activity and biodistribution of polypeptide K237 and folic acid conjugated amphiphilic PEG-PLGA copolymer nanoparticles radiolabeled with 99mTc
- Author
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Hao Jiang, Zelai He, Jingwen Huang, Jing Ma, Xuanzhang Huang, Yufeng Wu, Jie Chen, Xiangyu Zhang, Junyong Xia, and Jian Wu
- Subjects
Male ,K237 ,Biodistribution ,medicine.medical_specialty ,Spleen ,02 engineering and technology ,Pharmacology ,folate ,Polyethylene Glycols ,Mice ,03 medical and health sciences ,Folic Acid ,0302 clinical medicine ,In vivo ,Cell Line, Tumor ,Neoplasms ,medicine ,Percent Injected Dose ,Animals ,Humans ,Tissue Distribution ,Receptor ,Polyglactin 910 ,biodistribution ,business.industry ,technology, industry, and agriculture ,Cancer ,Organotechnetium Compounds ,021001 nanoscience & nanotechnology ,medicine.disease ,Xenograft Model Antitumor Assays ,Surgery ,Disease Models, Animal ,medicine.anatomical_structure ,Oncology ,Folate receptor ,Isotope Labeling ,030220 oncology & carcinogenesis ,Nanoparticles ,0210 nano-technology ,Drug carrier ,business ,Oligopeptides ,PEG-PLGA ,Research Paper - Abstract
// Zelai He 1, 3, * , Xiangyu Zhang 4, * , Jingwen Huang 1, 3, * , Yufeng Wu 5 , Xuanzhang Huang 1 , Jie Chen 1 , Junyong Xia 6 , Hao Jiang 3 , Jing Ma 2 , Jian Wu 1 1 The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou, China 2 Ultrasonic Department, Shanghai Songjiang Center Hospital, Shanghai, China 3 The First Affiliated Hospital of Bengbu Medical College, Bengbu, China 4 Department of Pathology, Jining No.1 Peoples’ Hospital, Jining, China 5 Department of Internal Medicine, Affiliated Cancer Hospital of Zhengzhou University, Henan Cancer Hospital, Zhengzhou, China 6 Department of Nuclear Medicine, The Affiliated Provincial Hospital of Anhui Medical University, Hefei, China * These authors contributed equally to this work Correspondence to: Junyong Xia, email: xiabingbing@126.com Hao Jiang, email: jianghao1223@163.com Jing Ma, email: majing0709@163.com Jian Wu, email: wzzjian@hotmail.com Keywords: K237, folate, PEG-PLGA, nanoparticles, biodistribution Received: July 29, 2016 Accepted: October 12, 2016 Published: October 24, 2016 ABSTRACT In a previous study, amphiphilic copolymer, polypeptide K237 (HTMYYHHYQHHL) and folic acid (FA) modified poly(ethylene glycol)-poly(lactic-co-glycolic acid) (K237/FA-PEG-PLGA) nanoparticles were developed and studied as a drug carrier. To further promote the clinical application of K237/FA-PEG-PLGA nanoparticles and provide guidance for future research, we need to examine their specific biodistribution in vivo . In this study, K237/FA-PEG-PLGA nanoparticles were effectively labeled by a direct method with Technetium-99m ( 99m Tc) using stannous chloride as a reducing agent. The optimal stability of the labeled nanoparticles was determined by evaluating their radiochemical purity in serum, physiological saline, diethylenetriaminepentaacetic acid (DTPA) and cysteine solutions. The affinity of ligands and receptors was elicited by cell binding and blocking experiments in KDR/folate receptor high expressing SKOV-3 ovarian cancer cells. The nanoparticles biodistribution was studied after intravenous administration in healthy mice xenografted with SKOV-3 cells. A higher percent injected dose per gram of tissue (% ID/g) was observed in liver, kidney, spleen, blood and tumor at 3 and 9 h post-injection. Scintigraphic images revealed that the radioactivity was mainly concentrated in tumor, liver, kidney and bladder; and in the heart, lung, and muscle was significantly lower at 3 h. The radioactivity distribution in the images is consistent with the in vivo biodistribution data. Our works demonstrated that K237/FA-PEG-PLGA nanoparticles have great potential as biodegradable drug carriers, especially for tumors expressing the folate and KDr receptor.
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- 2016
49. Cannabinoids synergize with carfilzomib, reducing multiple myeloma cells viability and migration
- Author
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Pietro Leoni, Claudio Cardinali, Silvia Gentili, Consuelo Amantini, Giorgio Santoni, Maria Beatrice Morelli, Massimo Nabissi, Alessandra Soriani, and Massimo Offidani
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0301 basic medicine ,Integrin beta Chains ,THC ,Combination therapy ,Cell Survival ,Antineoplastic Agents ,Pharmacology ,CXCR4 ,combination therapy ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Cell Movement ,Cell Line, Tumor ,Autophagy ,medicine ,Humans ,Viability assay ,Tetrahydrocannabinol ,Cell Proliferation ,carfilzomib ,Dose-Response Relationship, Drug ,Cannabinoids ,Bortezomib ,business.industry ,Cell Cycle Checkpoints ,Carfilzomib ,multiple myeloma ,030104 developmental biology ,immuno-proteasome ,Oncology ,chemistry ,030220 oncology & carcinogenesis ,Proteasome inhibitor ,CBD ,business ,Oligopeptides ,Cannabidiol ,Biomarkers ,Research Paper ,medicine.drug - Abstract
// Massimo Nabissi 1, * , Maria Beatrice Morelli 2, * , Massimo Offidani 3 , Consuelo Amantini 4 , Silvia Gentili 3 , Alessandra Soriani 2 , Claudio Cardinali 2 , Pietro Leoni 3 , Giorgio Santoni 1 1 School of Pharmacy, Experimental Medicine Section, University of Camerino, Camerino, Italy 2 Department of Molecular Medicine, Sapienza University, Rome, Italy 3 Clinica di Ematologia, Azienda Ospedaliero Universitaria Ospedali Riuniti di Ancona, Ancona, Italy 4 School of Biosciences and Veterinary Medicine, University of Camerino, Camerino, Italy * These authors have contributed equally to this work Correspondence to: Massimo Nabissi, email: massimo.nabissi@unicam.it Keywords: carfilzomib, THC, CBD, multiple myeloma, immuno-proteasome, combination therapy Received: September 01, 2016 Accepted: October 05, 2016 Published: October 18, 2016 ABSTRACT Several studies showed a potential anti-tumor role for cannabinoids, by modulating cell signaling pathways involved in cancer cell proliferation, chemo-resistance and migration. Cannabidiol (CBD) was previously noted in multiple myeloma (MM), both alone and in synergy with the proteasome inhibitor bortezomib, to induce cell death. In other type of human cancers, the combination of CBD with Δ 9 -tetrahydrocannabinol (THC) was found to act synergistically with other chemotherapeutic drugs suggesting their use in combination therapy. In the current study, we evaluated the effects of THC alone and in combination with CBD in MM cell lines. We found that CBD and THC, mainly in combination, were able to reduce cell viability by inducing autophagic-dependent necrosis. Moreover, we showed that the CBD-THC combination was able to reduce MM cells migration by down-regulating expression of the chemokine receptor CXCR4 and of the CD147 plasma membrane glycoprotein. Furthermore, since the immuno-proteasome is considered a new target in MM and also since carfilzomib (CFZ) is a new promising immuno-proteasome inhibitor that creates irreversible adducts with the β5i subunit of immuno-proteasome, we evaluated the effect of CBD and THC in regulating the expression of the β5i subunit and their effect in combination with CFZ. Herein, we also found that the CBD and THC combination is able to reduce expression of the β5i subunit as well as to act in synergy with CFZ to increase MM cell death and inhibits cell migration. In summary, these results proved that this combination exerts strong anti-myeloma activities.
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- 2016
50. Synergistic targeting of Sp1, a critical transcription factor for myeloma cell growth and survival, by panobinostat and proteasome inhibitors
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Itsuro Endo, Asuko Oda, Ryota Amachi, Shingen Nakamura, Shiroh Fujii, Kumiko Kagawa, Masahiro Hiasa, Jumpei Teramachi, Sumiko Yoshida, Ariunzaya Bat-Erdene, Hirofumi Tenshin, Takeshi Harada, Kimiko Sogabe, Masahiro Abe, Masami Iwasa, Hirokazu Miki, and Ken-ichi Aihara
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0301 basic medicine ,panobinostat ,medicine.medical_specialty ,Indoles ,Cell Survival ,Sp1 Transcription Factor ,proteasome inhibitors ,Antineoplastic Agents ,Hydroxamic Acids ,caspase-8 ,Sp1 ,Bortezomib ,Cell therapy ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Cell Line, Tumor ,Panobinostat ,Internal medicine ,medicine ,Humans ,Multiple myeloma ,Cell Proliferation ,Hematology ,business.industry ,Drug Synergism ,medicine.disease ,Carfilzomib ,Up-Regulation ,multiple myeloma ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,Oncology ,chemistry ,Proteasome ,030220 oncology & carcinogenesis ,Immunology ,Proteasome inhibitor ,Cancer research ,business ,Oligopeptides ,Research Paper ,medicine.drug - Abstract
// Ariunzaya Bat-Erdene 1 , Hirokazu Miki 2 , Asuko Oda 1 , Shingen Nakamura 1 , Jumpei Teramachi 1, 4 , Ryota Amachi 1, 3 , Hirofumi Tenshin 1, 3 , Masahiro Hiasa 1, 3 , Masami Iwasa 1 , Takeshi Harada 1 , Shiro Fujii 1 , Kimiko Sogabe 1 , Kumiko Kagawa 1 , Sumiko Yoshida 1 , Itsuro Endo 1 , Kenichi Aihara 1 , Masahiro Abe 1 1 Department of Hematology, Endocrinology and Metabolism, Institute of Biomedical Sciences, Tokushima University Graduate School of Medicine, Tokushima, Japan 2 Division of Transfusion Medicine and Cell Therapy, Tokushima University Hospital, Tokushima, Japan 3 Department of Orthodontics and Dentofacial Orthopedics, Tokushima University Graduate School of Oral Sciences, Tokushima, Japan 4 Department of Histology and Oral Histology, Tokushima University Graduate School of Oral Sciences, Tokushima, Japan Correspondence to: Masahiro Abe, email: masabe@tokushima-u.ac.jp Keywords: multiple myeloma, panobinostat, proteasome inhibitors, caspase-8, Sp1 Received: July 16, 2016 Accepted: September 29, 2016 Published: October 12, 2016 ABSTRACT Panobinostat, a pan-deacetylase inhibitor, synergistically elicits cytotoxic activity against myeloma (MM) cells in combination with the proteasome inhibitor bortezomib. Because precise mechanisms for panobinostat’s anti-MM action still remain elusive, we aimed to clarify the mechanisms of anti-MM effects of panobinostat and its synergism with proteasome inhibitors. Although the transcription factor Sp1 was overexpressed in MM cells, the Sp1 inhibitor terameprocol induced MM cell death in parallel with reduction of IRF4 and cMyc. Panobinostat induced activation of caspase-8, which was inversely correlated with reduction of Sp1 protein levels in MM cells. The panobinostat-mediated effects were further potentiated to effectively induce MM cell death in combination with bortezomib or carfilzomib even at suboptimal concentrations as a single agent. Addition of the caspase-8 inhibitor z-IETD-FMK abolished the Sp1 reduction not only by panobinostat alone but also by its combination with bortezomib, suggesting caspase-8-mediated Sp1 degradation. The synergistic Sp1 reduction markedly suppressed Sp1-driven prosurvival factors, IRF4 and cMyc. Besides, the combinatory treatment reduced HDAC1, another Sp1 target, in MM cells, which may potentiate HDAC inhibition. Collectively, caspase-8-mediated post-translational Sp1 degradation appears to be among major mechanisms for synergistic anti-MM effects of panobinostat and proteasome inhibitors in combination.
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- 2016
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