35 results on '"Cruz-Garcia L"'
Search Results
2. Regulation of lipid metabolism and peroxisome proliferator-activated receptors in rainbow trout adipose tissue by lipolytic and antilipolytic endocrine factors
- Author
-
Cruz-Garcia, L., Sánchez-Gurmaches, J., Monroy, M., Gutiérrez, J., and Navarro, I.
- Published
- 2015
- Full Text
- View/download PDF
3. Insulin, IGF-I, and muscle MAPK pathway responses after sustained exercise and their contribution to growth and lipid metabolism regulation in gilthead sea bream
- Author
-
Sánchez-Gurmaches, J., Cruz-Garcia, L., Ibarz, A., Fernández-Borrás, J., Blasco, J., Gutiérrez, J., and Navarro, I.
- Published
- 2013
- Full Text
- View/download PDF
4. Tracking preleukemic cells in vivo to reveal the sequence of molecular events in radiation leukemogenesis
- Author
-
Verbiest, T, Finnon, P, Brown, N, Cruz-Garcia, L, O'Brien, G, Ross, E, Bouffler, S, Scudamore, C L, and Badie, C
- Abstract
Epidemiological studies have demonstrated an increased leukemia incidence following ionizing radiation exposure, but to date, the target cells and underlying mechanisms of radiation leukemogenesis remain largely unidentified. We engineered a mouse model carrying a different fluorescent marker on each chromosome 2, located inside the minimum deleted region occurring after radiation exposure and recognized as the first leukemogenic event. Using this tailored model, we report that following radiation exposure, more than half of asymptomatic CBA Sfpi1GFP/mCh mice presented with expanding clones of preleukemic hematopoietic cells harboring a hemizygous interstitial deletion of chromosome 2. Moreover, following isolation of preleukemic hematopoietic stem and progenitor cells irradiated in their native microenvironment, we identified the presence of Sfpi1 point mutations within a subpopulation of these preleukemic cells expanding rapidly (increasing from 6% to 55% in 21 days in peripheral blood in one case), hence identifying for the first time the presence of such cells within a living animal. Importantly, we also report a previously undescribed gender difference in the phenotype of the preleukemic cells and leukemia, suggesting a gender imbalance in the radiation-induced leukemic target cell. In conclusion, we provide novel insights into the sequence of molecular events occurring during the (radiation-induced) leukemic clonal evolution.
- Published
- 2018
5. Regulation of lipoprotein lipase gene expression by insulin and troglitazone in rainbow trout (Oncorhynchus mykiss) adipocyte cells in culture
- Author
-
Bouraoui, L., primary, Cruz-Garcia, L., additional, Gutiérrez, J., additional, Capilla, E., additional, and Navarro, I., additional
- Published
- 2012
- Full Text
- View/download PDF
6. Effect of dietary fish meal and fish oil replacement on lipogenic and lipoprotein lipase activities and plasma insulin in gilthead sea bream (Sparus aurata)
- Author
-
BOURAOUI, L., primary, SÁNCHEZ-GURMACHES, J., additional, CRUZ-GARCIA, L., additional, GUTIÉRREZ, J., additional, BENEDITO-PALOS, L., additional, PÉREZ-SÁNCHEZ, J., additional, and NAVARRO, I., additional
- Published
- 2010
- Full Text
- View/download PDF
7. Molecular cloning, tissue expression and regulation of liver X Receptor (LXR) transcription factors of Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss)
- Author
-
Cruz-Garcia, L., primary, Minghetti, M., additional, Navarro, I., additional, and Tocher, D.R., additional
- Published
- 2009
- Full Text
- View/download PDF
8. Effect of dietary fish meal and fish oil replacement on lipogenic and lipoprotein lipase activities and plasma insulin in gilthead sea bream ( Sparus aurata).
- Author
-
BOURAOUI, L., SÁNCHEZ-GURMACHES, J., CRUZ-GARCIA, L., GUTIÉRREZ, J., BENEDITO-PALOS, L., PÉREZ-SÁNCHEZ, J., and NAVARRO, I.
- Subjects
SPARUS aurata ,FISH feeds ,FISH meal as feed ,FISH oils as feed ,LIPOPROTEIN lipase ,INSULIN ,BLOOD plasma ,LIPID metabolism - Abstract
The effects of a double replacement of fish oil (FO) and fish meal (FM) by dietary vegetable ingredients in juvenile gilthead sea bream ( Sparus aurata L. 1758) on some indices of lipid metabolism and plasma insulin levels were analysed. Four experimental diets with a replacement of 75% of FM by plant proteins (PP) were administered. Added oil was either FO (75PP/FO diet), or a vegetable oil mix (VO), replacing 33%, 66% or 100% of FO (75PP/33VO, 75PP/66VO, 75PP/100VO diets). Another diet with 50% of substitution of FM by PP and with 100% of VO was also tested (50PP/100VO diet). Final body weight was similar in all diet groups, except for the 75PP/100VO group, which presented lower values. Circulating insulin levels increased with feed administration in all groups and no differences between diets were observed, with the exception of the 75PP/FO group, which presented higher plasma insulin values. In adipose tissue, glucose-6-phosphate dehydrogenase and malic enzyme activities decreased with the inclusion of vegetable oil, especially 5 h after feeding. Diet had no significant effect on the hepatic activity of either enzyme. Lipoprotein lipase activity decreased in white muscle and adipose tissue with the replacement of fish oil in 75PP diets, 5 h after feeding. In conclusion, the use of a combined replacement of fish oil and fish meal by vegetable ingredients in gilthead sea bream permits satisfactory growth, with moderate changes in tissue lipogenesis and lipid uptake. [ABSTRACT FROM AUTHOR]
- Published
- 2011
- Full Text
- View/download PDF
9. The value of may as an evidential and epistemic marker in English medical abstracts
- Author
-
Alonso-Almeida Francisco and Cruz-García Laura
- Subjects
English language ,PE1-3729 - Published
- 2011
- Full Text
- View/download PDF
10. Transcriptional Inflammatory Signature in Healthy Donors and Different Radiotherapy Cancer Patients.
- Author
-
O'Brien G, Kamuda M, Cruz-Garcia L, Polozova M, Tichy A, Markova M, Sirak I, Zahradnicek O, Widłak P, Ponge L, Polanska J, and Badie C
- Subjects
- Male, Humans, Lipopolysaccharides, Inflammation genetics, Radiation Oncology, Radiation Exposure, Prostatic Neoplasms
- Abstract
Cancer and ionizing radiation exposure are associated with inflammation. To identify a set of radiation-specific signatures of inflammation-associated genes in the blood of partially exposed radiotherapy patients, differential expression of 249 inflammatory genes was analyzed in blood samples from cancer patients and healthy individuals. The gene expression analysis on a cohort of 63 cancer patients (endometrial, head and neck, and prostate cancer) before and during radiotherapy (24 h, 48 h, ~1 week, ~4-8 weeks, and 1 month after the last fraction) identified 31 genes and 15 up- and 16 down-regulated genes. Transcription variability under normal conditions was determined using blood drawn on three separate occasions from four healthy donors. No difference in inflammatory expression between healthy donors and cancer patients could be detected prior to radiotherapy. Remarkably, repeated sampling of healthy donors revealed an individual endogenous inflammatory signature. Next, the potential confounding effect of concomitant inflammation was studied in the blood of seven healthy donors taken before and 24 h after a flu vaccine or ex vivo LPS (lipopolysaccharide) treatment; flu vaccination was not detected at the transcriptional level and LPS did not have any effect on the radiation-induced signature identified. Finally, we identified a radiation-specific signature of 31 genes in the blood of radiotherapy patients that were common for all cancers, regardless of the immune status of patients. Confirmation via MQRT-PCR was obtained for BCL6, MYD88, MYC, IL7, CCR4 and CCR7. This study offers the foundation for future research on biomarkers of radiation exposure, radiation sensitivity, and radiation toxicity for personalized radiotherapy treatment.
- Published
- 2024
- Full Text
- View/download PDF
11. Spi1 R235C point mutation confers hypersensitivity to radiation-induced acute myeloid leukemia in mice.
- Author
-
Brown N, Finnon R, Finnon P, McCarron R, Cruz-Garcia L, O'Brien G, Herbert E, Scudamore CL, Morel E, and Badie C
- Abstract
Ionizing radiation (IR) is a risk factor for acute myeloid leukemia (rAML). Murine rAMLs feature both hemizygous chromosome 2 deletions (Del2) and point mutations (R235) within the hematopoietic regulatory gene Spi1 . We generated a heterozygous CBA Spi1 R235 mouse (CBA
Spm/+ ) which develops de novo AML with 100% incidence by ∼12 months old and shows a dose-dependent reduction in latency following X-irradiation. These effects are reduced on an AML-resistant C57Bl6 genetic background. CBASpm/Gfp reporter mice show increased Gfp expression, indicating compensation for Spm-induced Spi1 haploinsufficiency. Del2 is always detected in both de novo and rAMLs, indicating that biallelic Spi1 mutation is required for AML. CBASpm/+ mice show that a single Spm modification is sufficient for initiating AML development with complete penetrance, via the "two-hit" mechanism and this is accelerated by IR exposure. Similar SPI1/PU.1 polymorphisms in humans could potentially lead to enhanced susceptibility to IR following medical or environmental exposure., Competing Interests: The Authors declare no competing interests., (Crown Copyright © 2023.)- Published
- 2023
- Full Text
- View/download PDF
12. The miRNA Content of Bone Marrow-Derived Extracellular Vesicles Contributes to Protein Pathway Alterations Involved in Ionising Radiation-Induced Bystander Responses.
- Author
-
Csordás IB, Rutten EA, Szatmári T, Subedi P, Cruz-Garcia L, Kis D, Jezsó B, Toerne CV, Forgács M, Sáfrány G, Tapio S, Badie C, and Lumniczky K
- Subjects
- Mice, Animals, Bone Marrow metabolism, Proteomics, Mice, Inbred CBA, Radiation, Ionizing, MicroRNAs genetics, MicroRNAs metabolism, Extracellular Vesicles metabolism
- Abstract
Extracellular vesicles (EVs), through their cargo, are important mediators of bystander responses in the irradiated bone marrow (BM). MiRNAs carried by EVs can potentially alter cellular pathways in EV-recipient cells by regulating their protein content. Using the CBA/Ca mouse model, we characterised the miRNA content of BM-derived EVs from mice irradiated with 0.1 Gy or 3 Gy using an nCounter analysis system. We also analysed proteomic changes in BM cells either directly irradiated or treated with EVs derived from the BM of irradiated mice. Our aim was to identify key cellular processes in the EV-acceptor cells regulated by miRNAs. The irradiation of BM cells with 0.1 Gy led to protein alterations involved in oxidative stress and immune and inflammatory processes. Oxidative stress-related pathways were also present in BM cells treated with EVs isolated from 0.1 Gy-irradiated mice, indicating the propagation of oxidative stress in a bystander manner. The irradiation of BM cells with 3 Gy led to protein pathway alterations involved in the DNA damage response, metabolism, cell death and immune and inflammatory processes. The majority of these pathways were also altered in BM cells treated with EVs from mice irradiated with 3 Gy. Certain pathways (cell cycle, acute and chronic myeloid leukaemia) regulated by miRNAs differentially expressed in EVs isolated from mice irradiated with 3 Gy overlapped with protein pathway alterations in BM cells treated with 3 Gy EVs. Six miRNAs were involved in these common pathways interacting with 11 proteins, suggesting the involvement of miRNAs in the EV-mediated bystander processes. In conclusion, we characterised proteomic changes in directly irradiated and EV-treated BM cells, identified processes transmitted in a bystander manner and suggested miRNA and protein candidates potentially involved in the regulation of these bystander processes.
- Published
- 2023
- Full Text
- View/download PDF
13. Deficient Radiation Transcription Response in COVID-19 Patients.
- Author
-
Polozov S, Cruz-Garcia L, O'Brien G, Goriacha V, Nasser F, Jeggo P, Candéias S, and Badie C
- Abstract
Purpose: The ongoing SARS-CoV-2 pandemic has resulted in over 6.3 million deaths and 560 million COVID-19 cases worldwide. Clinical management of hospitalized patients is complex due to the heterogeneous course of COVID-19. Low-dose radiation therapy is known to dampen localized chronic inflammation and has been suggested to be used to reduce lung inflammation in patients with COVID-19. However, it is unknown whether SARS-CoV-2 alters the radiation response and associated radiation exposure related risk., Methods and Materials: We generated gene expression profiles from circulating leukocytes of hospitalized patients with COVID-19 and healthy donors., Results: The p53 signaling pathway was found to be dysregulated, with mRNA levels of p53, ATM , and CHK2 being lower in patients with COVID-19. Several key p53 target genes involved in cell cycle arrest, apoptosis, and p53 feedback inhibition were upregulated in patients with COVID-19 while other p53 target genes were downregulated. This dysregulation has functional consequences as the transcription of p53 -dependant genes ( CCNG1, GADD45A, DDB2, SESN1, FDXR, APOBEC ) was reduced 24 hours after x-ray exposure ex vivo to both low (100 mGy) or high (2 Gy) doses., Conclusions: SARS-CoV-2 infection affects a DNA damage response that may modify radiation-induced health risks in exposed patients with COVID-19., (Crown Copyright © 2023 Published by Elsevier Inc. on behalf of American Society for Radiation Oncology.)
- Published
- 2023
- Full Text
- View/download PDF
14. Transcriptional Dynamics of DNA Damage Responsive Genes in Circulating Leukocytes during Radiotherapy.
- Author
-
Cruz-Garcia L, Nasser F, O'Brien G, Grepl J, Vinnikov V, Starenkiy V, Artiukh S, Gramatiuk S, and Badie C
- Abstract
External beam radiation therapy leads to cellular activation of the DNA damage response (DDR). DNA double-strand breaks (DSBs) activate the ATM/CHEK2/p53 pathway, inducing the transcription of stress genes. The dynamic nature of this transcriptional response has not been directly observed in vivo in humans. In this study we monitored the messenger RNA transcript abundances of nine DNA damage-responsive genes ( CDKN1A , GADD45 , CCNG1 , FDXR , DDB2 , MDM2 , PHPT1 , SESN1 , and PUMA ), eight of them regulated by p53 in circulating blood leukocytes at different time points (2, 6-8, 16-18, and 24 h) in cancer patients (lung, neck, brain, and pelvis) undergoing radiotherapy. We discovered that, although the calculated mean physical dose to the blood was very low (0.038-0.169 Gy), an upregulation of Ferredoxin reductase ( FDXR ) gene transcription was detectable 2 h after exposure and was dose dependent from the lowest irradiated percentage of the body (3.5% whole brain) to the highest, (up to 19.4%, pelvic zone) reaching a peak at 6-8 h. The radiation response of the other genes was not strong enough after such low doses to provide meaningful information. Following multiple fractions, the expression level increased further and was still significantly up-regulated by the end of the treatment. Moreover, we compared FDXR transcriptional responses to ionizing radiation (IR) in vivo with healthy donors' blood cells exposed ex vivo and found a good correlation in the kinetics of expression from the 8-hours time-point onward, suggesting that a molecular transcriptional regulation mechanism yet to be identified is involved. To conclude, we provided the first in vivo human report of IR-induced gene transcription temporal response of a panel of p53-dependant genes. FDXR was demonstrated to be the most responsive gene, able to reliably inform on the low doses following partial body irradiation of the patients, and providing an expression pattern corresponding to the % of body exposed. An extended study would provide individual biological dosimetry information and may reveal inter-individual variability to predict radiotherapy-associated adverse health outcomes.
- Published
- 2022
- Full Text
- View/download PDF
15. Oxidative Stress and X-ray Exposure Levels-Dependent Survival and Metabolic Changes in Murine HSPCs.
- Author
-
Karabulutoglu M, Finnon R, Cruz-Garcia L, Hill MA, and Badie C
- Abstract
Haematopoietic bone marrow cells are amongst the most sensitive to ionizing radiation (IR), initially resulting in cell death or genotoxicity that may later lead to leukaemia development, most frequently Acute Myeloid Leukaemia (AML). The target cells for radiation-induced Acute Myeloid Leukaemia (rAML) are believed to lie in the haematopoietic stem and progenitor cell (HSPC) compartment. Using the inbred strain CBA/Ca as a murine model of rAML, progress has been made in understanding the underlying mechanisms, characterisation of target cell population and responses to IR. Complex regulatory systems maintain haematopoietic homeostasis which may act to modulate the risk of rAML. However, little is currently known about the role of metabolic factors and diet in these regulatory systems and modification of the risk of AML development. This study characterises cellular proliferative and clonogenic potential as well as metabolic changes within murine HSPCs under oxidative stress and X-ray exposure. Ambient oxygen (normoxia; 20.8% O
2 ) levels were found to increase irradiated HSPC-stress, stimulating proliferative activity compared to low oxygen (3% O2 ) levels. IR exposure has a negative influence on the proliferative capability of HSPCs in a dose-dependent manner (0-2 Gy) and this is more pronounced under a normoxic state. One Gy x-irradiated HSPCs cultured under normoxic conditions displayed a significant increase in oxygen consumption compared to those cultured under low O2 conditions and to unirradiated HSPCs. Furthermore, mitochondrial analyses revealed a significant increase in mitochondrial DNA (mtDNA) content, mitochondrial mass and membrane potential in a dose-dependent manner under normoxic conditions. Our results demonstrate that both IR and normoxia act as stressors for HSPCs, leading to significant metabolic deregulation and mitochondrial dysfunctionality which may affect long term risks such as leukaemia.- Published
- 2021
- Full Text
- View/download PDF
16. An ionising radiation-induced specific transcriptional signature of inflammation-associated genes in whole blood from radiotherapy patients: a pilot study.
- Author
-
Cruz-Garcia L, Badie C, Anbalagan S, Moquet J, Gothard L, O'Brien G, Somaiah N, and Ainsbury EA
- Subjects
- Biomarkers, Tumor genetics, Breast Neoplasms blood, Breast Neoplasms genetics, Breast Neoplasms immunology, Breast Neoplasms radiotherapy, Female, Gastrointestinal Neoplasms blood, Gastrointestinal Neoplasms genetics, Gastrointestinal Neoplasms immunology, Gastrointestinal Neoplasms radiotherapy, Humans, Lung Neoplasms blood, Lung Neoplasms genetics, Lung Neoplasms immunology, Lung Neoplasms radiotherapy, Neoplasms genetics, Neoplasms immunology, Neoplasms radiotherapy, Pilot Projects, Prognosis, Urogenital Neoplasms blood, Urogenital Neoplasms genetics, Urogenital Neoplasms immunology, Urogenital Neoplasms radiotherapy, Biomarkers, Tumor blood, Gene Expression Regulation, Neoplastic radiation effects, Inflammation genetics, Neoplasms blood, Radiation, Ionizing, Transcriptome radiation effects
- Abstract
Background: This communication reports the identification of a new panel of transcriptional changes in inflammation-associated genes observed in response to ionising radiation received by radiotherapy patients., Methods: Peripheral blood samples were taken with ethical approval and informed consent from a total of 20 patients undergoing external beam radiotherapy for breast, lung, gastrointestinal or genitourinary tumours. Nanostring nCounter analysis of transcriptional changes was carried out in samples prior and 24 h post-delivery of the 1st radiotherapy fraction, just prior to the 5th or 6th fraction, and just before the last fraction., Results: Statistical analysis with BRB-ArrayTools, GLM MANOVA and nSolver, revealed a radiation responsive panel of genes which varied by patient group (type of cancer) and with time since exposure (as an analogue for dose received), which may be useful as a biomarker of radiation response., Conclusion: Further validation in a wider group of patients is ongoing, together with work towards a full understanding of patient specific responses in support of personalised approaches to radiation medicine.
- Published
- 2021
- Full Text
- View/download PDF
17. Role of blood derived cell fractions, temperature and sample transport on gene expression-based biological dosimetry.
- Author
-
Nasser F, Cruz-Garcia L, O'Brien G, and Badie C
- Abstract
Purpose: For triage purposes following a nuclear accident or a terrorist event, gene expression biomarkers in blood have been demonstrated to be good bioindicators of ionizing radiation (IR) exposure and can be used to assess the dose received by exposed individuals. Many IR-sensitive genes are regulated by the DNA damage response pathway, and modulators of this pathway could potentially affect their expression level and therefore alter accurate dose estimations. In the present study, we addressed the potential influence of temperature, sample transport conditions and the blood cell fraction analyzed on the transcriptional response of the following radiation-responsive genes: FDXR, CCNG1, MDM2, PHPT1, APOBEC3H, DDB2, SESN1, P21, PUMA, and GADD45., Materials and Methods: Whole blood from healthy donors was exposed to a 2 Gy X-ray dose with a dose rate of 0.5 Gy/min (output 13 mA, 250 kV peak, 0.2 mA) and incubated for 24 h at either 37, 22, or 4 °C. For mimicking the effect of transport conditions at different temperatures, samples incubated at 37 °C for 24 h were kept at 37, 22 or 4 °C for another 24 h. Comparisons of biomarker responses to IR between white blood cells (WBCs), peripheral blood mononuclear cells (PBMCs) and whole blood were carried out after a 2 Gy X-ray exposure and incubation at 37 °C for 24 hours., Results: Hypothermic conditions (22 or 4 °C) following irradiation drastically inhibited transcriptional responses to IR exposure. However, sample shipment at different temperatures did not affect gene expression level except for SESN1. The transcriptional response to IR of specific genes depended on the cell fraction used, apart from FDXR, CCNG1, and SESN1., Conclusion: In conclusion, temperature during the incubation period and cell fraction but not the storing conditions during transport can influence the transcriptional response of specific genes. However, FDXR and CCNG1 showed a consistent response under all the different conditions tested demonstrating their reliability as individual biological dosimetry biomarkers.
- Published
- 2021
- Full Text
- View/download PDF
18. In Vivo Validation of Alternative FDXR Transcripts in Human Blood in Response to Ionizing Radiation.
- Author
-
Cruz-Garcia L, O'Brien G, Sipos B, Mayes S, Tichý A, Sirák I, Davídková M, Marková M, Turner DJ, and Badie C
- Subjects
- Adult, Alternative Splicing, DNA Damage, Dose-Response Relationship, Radiation, Female, Gene Expression Regulation, Humans, Male, Middle Aged, Neoplasms radiotherapy, Radiation, Ionizing, Blood radiation effects, Neoplasms genetics, Oxidoreductases genetics, Up-Regulation
- Abstract
Following cell stress such as ionising radiation (IR) exposure, multiple cellular pathways are activated. We recently demonstrated that ferredoxin reductase (FDXR) has a remarkable IR-induced transcriptional responsiveness in blood. Here, we provided a first comprehensive FDXR variant profile following DNA damage. First, specific quantitative real-time polymerase chain reaction (qPCR) primers were designed to establish dose-responses for eight curated FDXR variants, all up-regulated after IR in a dose-dependent manner. The potential role of gender on the expression of these variants was tested, and neither the variants response to IR nor the background level of expression was profoundly affected; moreover, in vitro induction of inflammation temporarily counteracted IR response early after exposure. Importantly, transcriptional up-regulation of these variants was further confirmed in vivo in blood of radiotherapy patients. Full-length nanopore sequencing was performed to identify other FDXR variants and revealed the high responsiveness of FDXR-201 and FDXR-208. Moreover, FDXR-218 and FDXR-219 showed no detectable endogenous expression, but a clear detection after IR. Overall, we characterised 14 FDXR transcript variants and identified for the first time their response to DNA damage in vivo. Future studies are required to unravel the function of these splicing variants, but they already represent a new class of radiation exposure biomarkers.
- Published
- 2020
- Full Text
- View/download PDF
19. Kras mutations and PU.1 promoter methylation are new pathways in murine radiation-induced AML.
- Author
-
O'Brien G, Cruz-Garcia L, Zyla J, Brown N, Finnon R, Polanska J, and Badie C
- Subjects
- Animals, Carcinogenesis, DNA Methylation genetics, Down-Regulation, Humans, Mice, Mice, Inbred CBA, MicroRNAs genetics, Mutation, Promoter Regions, Genetic, fms-Like Tyrosine Kinase 3, Gene Expression Regulation, Leukemic, Leukemia, Experimental genetics, Leukemia, Myeloid, Acute genetics, Leukemia, Radiation-Induced genetics, Proto-Oncogene Proteins genetics, Proto-Oncogene Proteins p21(ras) genetics, Trans-Activators genetics
- Abstract
Therapy-related and more specifically radiotherapy-associated acute myeloid leukaemia (AML) is a well-recognized potential complication of cytotoxic therapy for the treatment of a primary cancer. The CBA mouse model is used to study radiation leukaemogenesis mechanisms with Sfpi1/PU.1 deletion and point mutation already identified as driving events during AML development. To identify new pathways, we analysed 123 mouse radiation-induced AML (rAML) samples for the presence of mutations identified previously in human AML and found three genes to be mutated; Sfpi1 R235 (68%), Flt3-ITD (4%) and Kras G12 (3%), of which G12R was previously unreported. Importantly, a significant decrease in Sfpi1 gene expression is found almost exclusively in rAML samples without an Sfpi1 R235 mutation and is specifically associated with up-regulation of mir-1983 and mir-582-5p. Moreover, this down-regulation of Sfpi1 mRNA is negatively correlated with DNA methylation levels at specific CpG sites upstream of the Sfpi1 transcriptional start site. The down regulation of Sfpi1/PU.1 has also been reported in human AML cases revealing one common pathway of myeloid disruption between mouse and human AML where dysregulation of Sfpi1/PU.1 is a necessary step in AML development., (© The Author(s) 2019. Published by Oxford University Press.)
- Published
- 2020
- Full Text
- View/download PDF
20. Generation of a Transcriptional Radiation Exposure Signature in Human Blood Using Long-Read Nanopore Sequencing.
- Author
-
Cruz-Garcia L, O'Brien G, Sipos B, Mayes S, Love MI, Turner DJ, and Badie C
- Subjects
- Dose-Response Relationship, Radiation, Genomics, Humans, Leukocytes, Mononuclear metabolism, Leukocytes, Mononuclear radiation effects, Blood metabolism, Blood radiation effects, Nanopore Sequencing, Radiation Exposure adverse effects, Transcription, Genetic radiation effects, Transcriptome radiation effects
- Abstract
In the event of a large-scale event leading to acute ionizing radiation exposure, high-throughput methods would be required to assess individual dose estimates for triage purposes. Blood-based gene expression is a broad source of biomarkers of radiation exposure which have great potential for providing rapid dose estimates for a large population. Time is a crucial component in radiological emergencies and the shipment of blood samples to relevant laboratories presents a concern. In this study, we performed nanopore sequencing analysis to determine if the technology can be used to detect radiation-inducible genes in human peripheral blood mononuclear cells (PBMCs). The technology offers not only long-read sequencing but also a portable device which can overcome issues involving sample shipment, and provide faster results. For this goal, blood from nine healthy volunteers was 2 Gy ex vivo X irradiated. After PBMC isolation, irradiated samples were incubated along with the controls for 24 h at 37°C. RNA was extracted, poly(A)+ enriched and reverse-transcribed before sequencing. The data generated was analyzed using a Snakemake pipeline modified to handle paired samples. The sequencing analysis identified a radiation signature consisting of 46 differentially expressed genes (DEGs) which included 41 protein-coding genes, a long non-coding RNA and four pseudogenes, five of which have been identified as radiation-responsive transcripts for the first time. The genes in which transcriptional expression is most significantly modified after radiation exposure were APOBEC3H and FDXR, presenting a 25- and 28-fold change on average, respectively. These levels of transcriptional response were comparable to results we obtained by quantitative polymerase chain reaction (qPCR) analysis. In vivo exposure analyses showed a transcriptional radioresponse at 24 h postirradiation for both genes together with a strong dose-dependent response in blood irradiated ex vivo . Finally, extrapolating from the data we obtained, the minimum sequencing time required to detect an irradiated sample using APOBEC3H transcripts would be less than 3 min for a total of 50,000 reads. Future improvements, in sample processing and bioinformatic pipeline for specific radiation-responsive transcript identification, will allow the provision of a portable, rapid, real-time biodosimetry platform based on this new sequencing technology. In summary, our data show that nanopore sequencing can identify radiation-responsive genes and can also be used for identification of new transcripts.
- Published
- 2020
- Full Text
- View/download PDF
21. RAPID GENE EXPRESSION BASED DOSE ESTIMATION FOR RADIOLOGICAL EMERGENCIES.
- Author
-
Polozov S, Cruz-Garcia L, and Badie C
- Subjects
- Adult, Dose-Response Relationship, Radiation, Emergencies, Female, Gene Expression Profiling, Humans, Male, Middle Aged, Radiation Injuries blood, Radiation Injuries genetics, X-Rays, Biomarkers analysis, Blood Proteins analysis, Gene Expression Regulation radiation effects, Radiation Injuries diagnosis, Transcriptome radiation effects, Triage methods
- Abstract
Gene expression (GE) assays have shown great potential for rapid individual radiation dose exposure assessment. The aim of the present study was to optimise GE-based biological dosimetry protocols for radiological emergencies. Experiments were carried out to validate a newly developed protocol (P2) where several steps were optimised and to compare it with the current validated protocol in place in our laboratory (P1). Several donor blood samples from were exposed ex vivo to of the following doses: 0, 0.5, 1, 2 Gy X-rays. Concomitant measurement of transcription level of genes FDXR, P21, PHPT1, CCNG1 and SESN1 plus HPRT (control) was performed. To summarise, both protocols provided similar dose estimates, P1 being completed in 7 hours while P2 in merely 4 hours. Thus, a significant time shortening was achieved leading to a potential increase of throughput capacity. Hence, this new protocol can be recommended for mass radiation casualties triage purposes., (© The Author(s) 2019. Published by Oxford University Press.)
- Published
- 2019
- Full Text
- View/download PDF
22. Hemocyanin facilitates lignocellulose digestion by wood-boring marine crustaceans.
- Author
-
Besser K, Malyon GP, Eborall WS, Paro da Cunha G, Filgueiras JG, Dowle A, Cruz Garcia L, Page SJ, Dupree R, Kern M, Gomez LD, Li Y, Elias L, Sabbadin F, Mohamad SE, Pesante G, Steele-King C, Ribeiro de Azevedo E, Polikarpov I, Dupree P, Cragg SM, Bruce NC, and McQueen-Mason SJ
- Subjects
- Animals, Cellulose metabolism, Diet, Digestion physiology, Feces chemistry, Gastrointestinal Tract metabolism, Gastrointestinal Tract ultrastructure, Isopoda metabolism, Microscopy, Electron, Scanning, Microscopy, Electron, Transmission, Xylans metabolism, Gastrointestinal Tract physiology, Hemocyanins metabolism, Isopoda physiology, Lignin metabolism, Wood parasitology
- Abstract
Woody (lignocellulosic) plant biomass is an abundant renewable feedstock, rich in polysaccharides that are bound into an insoluble fiber composite with lignin. Marine crustacean woodborers of the genus Limnoria are among the few animals that can survive on a diet of this recalcitrant material without relying on gut resident microbiota. Analysis of fecal pellets revealed that Limnoria targets hexose-containing polysaccharides (mainly cellulose, and also glucomannans), corresponding with the abundance of cellulases in their digestive system, but xylans and lignin are largely unconsumed. We show that the limnoriid respiratory protein, hemocyanin, is abundant in the hindgut where wood is digested, that incubation of wood with hemocyanin markedly enhances its digestibility by cellulases, and that it modifies lignin. We propose that this activity of hemocyanins is instrumental to the ability of Limnoria to feed on wood in the absence of gut symbionts. These findings may hold potential for innovations in lignocellulose biorefining.
- Published
- 2018
- Full Text
- View/download PDF
23. Dicentric Dose Estimates for Patients Undergoing Radiotherapy in the RTGene Study to Assess Blood Dosimetric Models and the New Bayesian Method for Gradient Exposure.
- Author
-
Moquet J, Higueras M, Donovan E, Boyle S, Barnard S, Bricknell C, Sun M, Gothard L, O'Brien G, Cruz-Garcia L, Badie C, Ainsbury E, and Somaiah N
- Subjects
- Adult, Aged, Bayes Theorem, Biomarkers, Tumor genetics, Breast Neoplasms blood, Breast Neoplasms pathology, Breast Neoplasms radiotherapy, Chromosomes radiation effects, Dose-Response Relationship, Radiation, Female, Gastrointestinal Neoplasms blood, Gastrointestinal Neoplasms pathology, Gastrointestinal Neoplasms radiotherapy, Humans, Lung Neoplasms blood, Lung Neoplasms pathology, Lung Neoplasms radiotherapy, Male, Middle Aged, Radiation Dosage, Radiation, Ionizing, Radiometry, Urogenital Neoplasms blood, Urogenital Neoplasms pathology, Urogenital Neoplasms radiotherapy, Biomarkers, Tumor blood, Chromosome Aberrations radiation effects, Chromosomes genetics
- Abstract
The RTGene study was focused on the development and validation of new transcriptional biomarkers for prediction of individual radiotherapy patient responses to ionizing radiation. In parallel, for validation purposes, this study incorporated conventional biomarkers of radiation exposure, including the dicentric assay. Peripheral blood samples were taken with ethical approval and informed consent from a total of 20 patients undergoing external beam radiotherapy for breast, lung, gastrointestinal or genitourinary tumors. For the dicentric assay, two samples were taken from each patient: prior to radiotherapy and before the final fraction. Blood samples were set up using standard methods for the dicentric assay. All the baseline samples had dicentric frequencies consistent with the expected background for the normal population. For blood taken before the final fraction, all the samples displayed distributions of aberrations, which are indicative of partial-body exposures. Whole-body and partial-body cytogenetic doses were calculated with reference to a 250-kVp X-ray calibration curve and then compared to the dose to blood derived using two newly developed blood dosimetric models. Initial comparisons indicated that the relationship between these measures of dose appear very promising, with a correlation of 0.88 ( P = 0.001). A new Bayesian zero-inflated Poisson finite mixture method was applied to the dicentric data, and partial-body dose estimates showed no significant difference ( P > 0.999) from those calculated by the contaminated Poisson technique. The next step will be further development and validation in a larger patient group.
- Published
- 2018
- Full Text
- View/download PDF
24. Influence of Confounding Factors on Radiation Dose Estimation Using In Vivo Validated Transcriptional Biomarkers.
- Author
-
Cruz-Garcia L, O'Brien G, Donovan E, Gothard L, Boyle S, Laval A, Testard I, Ponge L, Woźniak G, Miszczyk L, Candéias SM, Ainsbury E, Widlak P, Somaiah N, and Badie C
- Subjects
- Adult, Aged, Aged, 80 and over, Antineoplastic Agents pharmacology, Case-Control Studies, Curcumin pharmacology, Female, Gene Expression Regulation, Neoplastic drug effects, Humans, Lipopolysaccharides pharmacology, Male, Middle Aged, Neoplasms drug therapy, Neoplasms genetics, Neoplasms radiotherapy, Biomarkers blood, Cyclin G1 genetics, Gene Expression Regulation, Neoplastic radiation effects, Neoplasms blood, Phosphoric Monoester Hydrolases genetics, Radiotherapy Dosage standards, Radiotherapy, Intensity-Modulated
- Abstract
For triage purposes following a nuclear accident, blood-based gene expression biomarkers can provide rapid dose estimates for a large number of individuals. Ionizing-radiation-responsive genes are regulated through the DNA damage-response pathway, which includes activation of multiple transcription factors. Modulators of this pathway could potentially affect the response of these biomarkers and consequently compromise accurate dose estimation calculations. In the present study, four potential confounding factors were selected: cancer condition, sex, simulated bacterial infection (lipopolysaccharide), and curcumin, an anti-inflammatory/antioxidant agent. Their potential influence on the transcriptional response to radiation of the genes CCNG1 and PHPT1, two biomarkers of radiation exposure ex vivo, was assessed. First, both CCNG1 and PHPT1 were detected in vivo in blood samples from radiotherapy patients and as such were validated as biomarkers of exposure. Importantly, their basal expression level was slightly but significantly affected in vivo by patients' cancer condition. Moreover, lipopolysaccharide stimulation of blood irradiated ex vivo led to a significant modification of CCNG1 and PHPT1 transcriptional response in a dose- and time-dependent manner with opposite regulatory effects. Curcumin also affected CCNG1 and PHPT1 transcriptional response counteracting some of the radiation induction. No differences were observed based on sex. Dose estimations calculated using linear regression were affected by lipopolysaccharide and curcumin. In conclusion, several confounding factors tested in this study can indeed modulate the transcriptional response of CCNG1 and PHPT1 and consequently can affect radiation exposure dose estimations but not to a level which should prevent the biomarkers' use for triage purposes.
- Published
- 2018
- Full Text
- View/download PDF
25. Tracking preleukemic cells in vivo to reveal the sequence of molecular events in radiation leukemogenesis.
- Author
-
Verbiest T, Finnon R, Brown N, Cruz-Garcia L, Finnon P, O'Brien G, Ross E, Bouffler S, Scudamore CL, and Badie C
- Subjects
- Animals, Clonal Evolution, Disease Progression, Female, Hematopoietic Stem Cells radiation effects, Male, Mice, Mice, Inbred CBA, Point Mutation, Proto-Oncogene Proteins genetics, Sex Characteristics, Trans-Activators genetics, Leukemia, Radiation-Induced etiology, Preleukemia etiology
- Abstract
Epidemiological studies have demonstrated an increased leukemia incidence following ionizing radiation exposure, but to date, the target cells and underlying mechanisms of radiation leukemogenesis remain largely unidentified. We engineered a mouse model carrying a different fluorescent marker on each chromosome 2, located inside the minimum deleted region occurring after radiation exposure and recognized as the first leukemogenic event. Using this tailored model, we report that following radiation exposure, more than half of asymptomatic CBA Sfpi1
GFP/mCh mice presented with expanding clones of preleukemic hematopoietic cells harboring a hemizygous interstitial deletion of chromosome 2. Moreover, following isolation of preleukemic hematopoietic stem and progenitor cells irradiated in their native microenvironment, we identified the presence of Sfpi1 point mutations within a subpopulation of these preleukemic cells expanding rapidly (increasing from 6% to 55% in 21 days in peripheral blood in one case), hence identifying for the first time the presence of such cells within a living animal. Importantly, we also report a previously undescribed gender difference in the phenotype of the preleukemic cells and leukemia, suggesting a gender imbalance in the radiation-induced leukemic target cell. In conclusion, we provide novel insights into the sequence of molecular events occurring during the (radiation-induced) leukemic clonal evolution.- Published
- 2018
- Full Text
- View/download PDF
26. FDXR is a biomarker of radiation exposure in vivo.
- Author
-
O'Brien G, Cruz-Garcia L, Majewski M, Grepl J, Abend M, Port M, Tichý A, Sirak I, Malkova A, Donovan E, Gothard L, Boyle S, Somaiah N, Ainsbury E, Ponge L, Slosarek K, Miszczyk L, Widlak P, Green E, Patel N, Kudari M, Gleeson F, Vinnikov V, Starenkiy V, Artiukh S, Vasyliev L, Zaman A, and Badie C
- Subjects
- Adult, Aged, Aged, 80 and over, Curcumin pharmacology, Female, Ferredoxin-NADP Reductase genetics, Humans, Lipopolysaccharides pharmacology, Male, Middle Aged, Neoplasms metabolism, Neoplasms radiotherapy, RNA blood, RNA drug effects, Tomography, X-Ray Computed, Up-Regulation drug effects, Young Adult, Biomarkers metabolism, Ferredoxin-NADP Reductase metabolism, Whole-Body Irradiation
- Abstract
Previous investigations in gene expression changes in blood after radiation exposure have highlighted its potential to provide biomarkers of exposure. Here, FDXR transcriptional changes in blood were investigated in humans undergoing a range of external radiation exposure procedures covering several orders of magnitude (cardiac fluoroscopy, diagnostic computed tomography (CT)) and treatments (total body and local radiotherapy). Moreover, a method was developed to assess the dose to the blood using physical exposure parameters. FDXR expression was significantly up-regulated 24 hr after radiotherapy in most patients and continuously during the fractionated treatment. Significance was reached even after diagnostic CT 2 hours post-exposure. We further showed that no significant differences in expression were found between ex vivo and in vivo samples from the same patients. Moreover, potential confounding factors such as gender, infection status and anti-oxidants only affect moderately FDXR transcription. Finally, we provided a first in vivo dose-response showing dose-dependency even for very low doses or partial body exposure showing good correlation between physically and biologically assessed doses. In conclusion, we report the remarkable responsiveness of FDXR to ionising radiation at the transcriptional level which, when measured in the right time window, provides accurate in vivo dose estimates.
- Published
- 2018
- Full Text
- View/download PDF
27. Lxr-driven enterocyte lipid droplet formation delays transport of ingested lipids.
- Author
-
Cruz-Garcia L and Schlegel A
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Biological Transport, Caco-2 Cells, Coenzyme A Ligases genetics, Coenzyme A Ligases metabolism, Diet, High-Fat adverse effects, Fatty Acids metabolism, Fatty Liver etiology, Fatty Liver metabolism, Female, Gene Expression, Humans, Hypercholesterolemia etiology, Hypercholesterolemia metabolism, Intestinal Absorption, Lipid Metabolism, Liver X Receptors, Male, Molecular Sequence Data, Zebrafish, Enterocytes metabolism, Lipid Droplets metabolism, Orphan Nuclear Receptors physiology
- Abstract
Liver X receptors (Lxrs) are master regulators of cholesterol catabolism, driving the elimination of cholesterol from the periphery to the lumen of the intestine. Development of pharmacological agents to activate Lxrs has been hindered by synthetic Lxr agonists' induction of hepatic lipogenesis and hypertriglyceridemia. Elucidating the function of Lxrs in regulating enterocyte lipid handling might identify novel aspects of lipid metabolism that are pharmacologically amenable. We took a genetic approach centered on the single Lxr gene nr1h3 in zebrafish to study the role of Lxr in enterocyte lipid metabolism. Loss of nr1h3 function causes anticipated gene regulatory changes and cholesterol intolerance, collectively reflecting high evolutionary conservation of zebrafish Lxra function. Intestinal nr1h3 activation delays transport of absorbed neutral lipids, with accumulation of neutral lipids in enterocyte cytoplasmic droplets. This delay in transport of ingested neutral lipids protects animals from hypercholesterolemia and hepatic steatosis induced by a high-fat diet. On a gene regulatory level, Lxra induces expression of acsl3a, which encodes acyl-CoA synthetase long-chain family member 3a, a lipid droplet-anchored protein that directs fatty acyl chains into lipids. Forced overexpression of acls3a in enterocytes delays, in part, the appearance of neutral lipids in the vasculature of zebrafish larvae. Activation of Lxr in the intestine cell-autonomously regulates the rate of delivery of absorbed lipids by inducting a temporary lipid intestinal droplet storage depot., (Copyright © 2014 by the American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2014
- Full Text
- View/download PDF
28. mRNA expression of fatty acid transporters in rainbow trout: in vivo and in vitro regulation by insulin, fasting and inflammation and infection mediators.
- Author
-
Sánchez-Gurmaches J, Cruz-Garcia L, Gutiérrez J, and Navarro I
- Subjects
- Adipocytes metabolism, Adipose Tissue immunology, Adipose Tissue metabolism, Animals, CD36 Antigens genetics, Cell Differentiation, Cells, Cultured, Fish Proteins genetics, Food Deprivation physiology, Inflammation Mediators pharmacology, Insulin-Like Growth Factor I pharmacology, Insulin-Like Growth Factor I physiology, Lipopolysaccharides pharmacology, Liver immunology, Liver metabolism, Muscle Fibers, Fast-Twitch immunology, Muscle Fibers, Fast-Twitch metabolism, Muscle Fibers, Slow-Twitch immunology, Muscle Fibers, Slow-Twitch metabolism, Myocardium metabolism, Organ Specificity, RNA, Messenger genetics, Satellite Cells, Skeletal Muscle metabolism, Satellite Cells, Skeletal Muscle physiology, Tumor Necrosis Factor-alpha pharmacology, Tumor Necrosis Factor-alpha physiology, CD36 Antigens metabolism, Fish Proteins metabolism, Inflammation Mediators physiology, Insulin physiology, Oncorhynchus mykiss metabolism, RNA, Messenger metabolism
- Abstract
In the present study, we analyzed endocrine and nutritional regulation of fatty acid (FA) transporters mRNA expression fatty acid transport protein (FATP1) and fatty acid translocase (CD36) in rainbow trout in vivo and in adipocytes and myocytes in vitro. The expression of FATP1 increased with adipocyte and that of CD36 with myocyte in vitro differentiation suggesting a different role for each transporter during the two cell differentiation programs. Food deprivation (15, 25 and 35 days) increased FATP1 and CD36 mRNA expression in white muscle, red muscle and adipose tissue while insulin administration decreased the FATP1 expression in adipose tissue in vivo (21.6 pmol/g body mass) and in vitro (1 μM) in adipocytes. In trout myotubes insulin (1 μM) decreased FATP1 and increased CD36 mRNA expression. Thus, regulation of FA transporters expression by insulin is complex and directed to specific tissue needs. Although FATP1 and CD36 mRNA levels are controlled by insulin, it appears that FATP1 respond more clearly to situations of hyper and hypo-insulinemia in trout muscle and adipose tissue than CD36. FATP1 and CD36 transcription was also modulated by growth hormone in cultured myotubes and isolated adipocytes. Lipopolysaccharide administration (E. coli, serotype O26:B6, 6 μg/g body mass) decreased FATP1 mRNA expression in red muscle, adipose tissue and liver after 24h according to changes in lipid metabolism during infection. Tumor necrosis factor (TNFα) (100 ng/ml) reduced FATP1 expression in isolated adipocytes. Further, insulin (1μM) and IGF-I (100 nM) increased the FA uptake in rainbow trout myotubes through the PI3K/Akt signaling pathway. Overall, we demonstrated not only that feeding condition regulates FATP1 and CD36 mRNA expression in a tissue-specific manner, but also that insulin is an important regulator of these genes in vivo and in vitro and also it stimulates FA uptake in trout muscle cells., (Copyright © 2012 Elsevier Inc. All rights reserved.)
- Published
- 2012
- Full Text
- View/download PDF
29. Role of LXR in trout adipocytes: target genes, hormonal regulation, adipocyte differentiation and relation to lipolysis.
- Author
-
Cruz-Garcia L, Sánchez-Gurmaches J, Gutiérrez J, and Navarro I
- Subjects
- Adipocytes cytology, Animals, Cell Differentiation, Liver X Receptors, Oncorhynchus mykiss genetics, Orphan Nuclear Receptors agonists, Orphan Nuclear Receptors genetics, RNA, Messenger agonists, RNA, Messenger genetics, RNA, Messenger metabolism, Adipocytes metabolism, Lipolysis, Oncorhynchus mykiss metabolism, Orphan Nuclear Receptors metabolism
- Abstract
In the present study, we describe an initial approach to investigate the role of LXR in fish adipose tissue. Rainbow trout (Oncorhynchus mykiss) isolated adipocytes were incubated with LXR agonists, unsaturated fatty acids, tumour necrosis factor-α (TNFα), insulin or growth hormone (GH) for 6h and LXR expression was analyzed. Lipolysis was measured after incubation with one of the LXR agonists and LXR expression was compared with levels of lipolysis. LXR expression was also analyzed during the differentiation of adipocytes in culture. The incubations with agonists in isolated adipocytes indicated that ATP-binding cassette transporter A1 (ABCA1) is an LXR target gene, but lipoprotein lipase (LPL), fatty acid synthase (FAS), hormone-sensitive lipase (HSL) and peroxisome proliferator-activated receptor (PPARs) are not. LXR agonists also induced LXR expression and raised lipolysis levels. Besides, LXR expression was upregulated in parallel with basal lipolysis. LXR mRNA expression was regulated by unsaturated fatty acids, insulin, TNFα and GH in isolated adipocytes. Besides, LXR showed an upregulation during adipocyte differentiation. All these data indicate that LXR is involved in orchestrating the transcriptional regulatory network in trout adipocyte lipid metabolism, specifically, in cholesterol transport, adipocyte differentiation and lipolysis., (Copyright © 2012 Elsevier Inc. All rights reserved.)
- Published
- 2012
- Full Text
- View/download PDF
30. Adiponectin effects and gene expression in rainbow trout: an in vivo and in vitro approach.
- Author
-
Sánchez-Gurmaches J, Cruz-Garcia L, Gutiérrez J, and Navarro I
- Subjects
- Adipocytes cytology, Adipocytes drug effects, Adipocytes metabolism, Adiponectin metabolism, Animals, Cell Differentiation drug effects, Cell Differentiation genetics, Fasting, Fatty Acids metabolism, Gene Expression Profiling, Glucose metabolism, Humans, Injections, Insulin administration & dosage, Insulin pharmacology, Lipopolysaccharides administration & dosage, Lipopolysaccharides pharmacology, Muscle Fibers, Skeletal drug effects, Muscle Fibers, Skeletal metabolism, Muscle, Skeletal cytology, RNA, Messenger genetics, RNA, Messenger metabolism, Receptors, Adiponectin genetics, Receptors, Adiponectin metabolism, Signal Transduction drug effects, Signal Transduction genetics, Tissue Distribution drug effects, Tumor Necrosis Factor-alpha administration & dosage, Tumor Necrosis Factor-alpha pharmacology, Adiponectin genetics, Gene Expression Regulation drug effects, Oncorhynchus mykiss genetics
- Abstract
Here we present the presence of adiponectin and adiponectin receptors [type 1 (adipoR1) and type 2 (adipoR2)] in rainbow trout (Oncorhynchus mykiss) tissues and cell cultures together with the response to different scenarios. In response to fasting, adiponectin expression was up-regulated in adipose tissue, while the expression of its receptors increased in white and red muscle. Insulin injection decreased adipoR1 expression in white and red muscles. We deduce that the adipoRs in trout muscle show opposite responses to increasing insulin plasma levels, which may maintain sensitivity to insulin in this tissue. Adiponectin expression was inhibited by the inflammatory effect of lipopolysaccharide (LPS) in adipose tissue and red muscle. Moreover, results indicate that LPS may lead to mobilization of fat reserves, increasing adipoR1 expression in adipose tissue. The effects of LPS could be mediated through tumour necrosis factor α (TNFα), at least in red muscle. Insulin, growth hormone and TNFα all diminished expression of adipoR2 in adipocytes and adipoR1 in myotubes, while insulin increased the expression of adipoR2 in the muscle cells. Adiponectin activates Akt in rainbow trout myotubes, which may lead to an increase in fatty acid uptake and oxidation. Overall, our results show that the adiponectin system responds differently to various physiological challenges and that it is hormonally controlled in vivo and in vitro. To the best of our knowledge, this is the first time this has been demonstrated in teleosts, and it may be a valuable contribution to our understanding of adipokines in fish.
- Published
- 2012
- Full Text
- View/download PDF
31. A monocarboxylate transporter required for hepatocyte secretion of ketone bodies during fasting.
- Author
-
Hugo SE, Cruz-Garcia L, Karanth S, Anderson RM, Stainier DY, and Schlegel A
- Subjects
- Animals, Embryo, Nonmammalian, Fatty Liver genetics, Fatty Liver pathology, Gene Expression Profiling, Gene Expression Regulation, Developmental, Humans, Larva, Monocarboxylic Acid Transporters genetics, Xenopus, Zebrafish, Fasting metabolism, Hepatocytes metabolism, Ketone Bodies metabolism, Monocarboxylic Acid Transporters metabolism
- Abstract
To find new genes that influence liver lipid mass, we performed a genetic screen for zebrafish mutants with hepatic steatosis, a pathological accumulation of fat. The red moon (rmn) mutant develops hepatic steatosis as maternally deposited yolk is depleted. Conversely, hepatic steatosis is suppressed in rmn mutants by adequate nutrition. Adult rmn mutants show increased liver neutral lipids and induction of hepatic lipid biosynthetic genes when fasted. Positional cloning of the rmn locus reveals a loss-of-function mutation in slc16a6a (solute carrier family 16a, member 6a), a gene that we show encodes a transporter of the major ketone body β-hydroxybutyrate. Restoring wild-type zebrafish slc16a6a expression or introducing human SLC16A6 in rmn mutant livers rescues the mutant phenotype. Radiotracer analysis confirms that loss of Slc16a6a function causes diversion of liver-trapped ketogenic precursors into triacylglycerol. Underscoring the importance of Slc16a6a to normal fasting physiology, previously fed rmn mutants are more sensitive to death by starvation than are wild-type larvae. Our unbiased, forward genetic approach has found a heretofore unrecognized critical step in fasting energy metabolism: hepatic ketone body transport. Since β-hydroxybutyrate is both a major fuel and a signaling molecule in fasting, the discovery of this transporter provides a new direction for modulating circulating levels of ketone bodies in metabolic diseases.
- Published
- 2012
- Full Text
- View/download PDF
32. Regulation of LXR by fatty acids, insulin, growth hormone and tumor necrosis factor-α in rainbow trout myocytes.
- Author
-
Cruz-Garcia L, Sánchez-Gurmaches J, Gutiérrez J, and Navarro I
- Subjects
- Animals, Cells, Cultured, Fish Proteins genetics, Fish Proteins metabolism, Gene Expression Profiling, Gene Expression Regulation, Developmental, Hydroxycholesterols pharmacology, Lipid Metabolism genetics, Liver X Receptors, Muscle Development genetics, Oncorhynchus mykiss genetics, Oncorhynchus mykiss growth & development, Orphan Nuclear Receptors agonists, Fatty Acids metabolism, Growth Hormone metabolism, Insulin metabolism, Muscle Fibers, Skeletal metabolism, Oncorhynchus mykiss metabolism, Orphan Nuclear Receptors metabolism, Tumor Necrosis Factor-alpha metabolism
- Abstract
The liver X receptor (LXR) has recently been described in salmonids. In mammals, this receptor is already known as a transcriptional factor that regulates diverse aspects of cholesterol, fatty acid and carbohydrate metabolism in various tissues, including muscle. Here we examined LXR in trout myocytes. For this purpose, we analyzed LXR target gene expression and gene profile during myocyte development. In addition, we studied the transcriptional regulation of LXR by hormones, a pro-inflammatory mediator and unsaturated fatty acids. Trout myocytes were incubated with LXR agonists (T091317, 22(R)-hydroxycholesterol) and unsaturated fatty acids for 24h. Furthermore, differentiated myocytes were incubated with insulin and growth hormone (GH) for 3h, 6h and 18h, and with tumor necrosis factor-α (TNFα) for 24h. Samples were also obtained in various stages of cell differentiation. Our results demonstrate that lipoprotein lipase (LPL), fatty acid synthase (FAS), ATP-binding cassette transporter A1 (ABCA1), peroxisome proliferator-activated receptor-α and β (PPARα, β) are target genes of LXR in trout muscle. LXR agonists increased LXR expression, thereby indicating that this receptor is autoregulated. Unsaturated fatty acids downregulated LXR gene expression. This observation suggests a regulatory mechanism of these molecules on LXR-mediated fatty acid synthesis and uptake. TNFα did not modulate LXR gene transcription. Expression of the LXR gene was activated by insulin and GH. These results suggest that LXR may play a lipogenic role through insulin stimulation and a tendency to promote anabolic effects through GH on trout myocytes., (Copyright © 2011 Elsevier Inc. All rights reserved.)
- Published
- 2011
- Full Text
- View/download PDF
33. Changes in adipocyte cell size, gene expression of lipid metabolism markers, and lipolytic responses induced by dietary fish oil replacement in gilthead sea bream (Sparus aurata L.).
- Author
-
Cruz-Garcia L, Sánchez-Gurmaches J, Bouraoui L, Saera-Vila A, Pérez-Sánchez J, Gutiérrez J, and Navarro I
- Subjects
- Adipocytes cytology, Animals, Biomarkers metabolism, Cell Size, Diet, Fish Oils administration & dosage, Gene Expression, Glucose metabolism, Insulin pharmacology, Lipase genetics, Lipase metabolism, Lipid Metabolism genetics, Lipolysis, Liver X Receptors, Orphan Nuclear Receptors genetics, Orphan Nuclear Receptors metabolism, Plant Oils administration & dosage, Sea Bream genetics, Tumor Necrosis Factor-alpha pharmacology, Adipocytes metabolism, Sea Bream metabolism
- Abstract
The effects of fish oil (FO) substitution by 66% vegetable oils in a diet with already 75% vegetable protein (66VO) on adipose tissue lipid metabolism of gilthead sea bream were analysed after a 14-month feeding trial. In the last 3 months of the experiment, a FO diet was administrated to a 66VO group (group 66VO/FO) as a finishing diet. Hormone-sensitive lipase (HSL) activity was measured in adipose tissue and adipocyte size, and HSL, lipoprotein lipase and liver X receptor gene expression in isolated adipocytes, on which lipolysis and glucose uptake experiments were also performed. Lipolysis was measured after incubation with tumour necrosis factor-α (TNFα), linoleic acid, and two conjugated linoleic acid isomers. Glucose uptake was analysed after TNFα or insulin administration. Our results show that FO replacement increased lipolytic activity and adipocyte cell size. The higher proportion of large cells observed in the 66VO group could be involved in their observed lower response to fatty acid treatments and lower insulin sensitivity. The 66VO/FO group showed a moderate return to the FO conditions. Therefore, FO replacement can affect the morphology and metabolism of gilthead sea bream adipocytes which could potentially affect other organs such as the liver., (Copyright © 2010 Elsevier Inc. All rights reserved.)
- Published
- 2011
- Full Text
- View/download PDF
34. Endocrine control of oleic acid and glucose metabolism in rainbow trout (Oncorhynchus mykiss) muscle cells in culture.
- Author
-
Sánchez-Gurmaches J, Cruz-Garcia L, Gutiérrez J, and Navarro I
- Subjects
- Animals, Carbon Dioxide metabolism, Carnitine O-Palmitoyltransferase antagonists & inhibitors, Carnitine O-Palmitoyltransferase metabolism, Catecholamines metabolism, Cells, Cultured, Epoxy Compounds pharmacology, Glycogen metabolism, Hypoglycemic Agents pharmacology, Intracellular Signaling Peptides and Proteins antagonists & inhibitors, Intracellular Signaling Peptides and Proteins metabolism, Muscle Fibers, Skeletal drug effects, Oxidation-Reduction, PPAR gamma agonists, PPAR gamma metabolism, Protein Kinase Inhibitors pharmacology, Protein Serine-Threonine Kinases antagonists & inhibitors, Protein Serine-Threonine Kinases metabolism, Rosiglitazone, Sirolimus pharmacology, TOR Serine-Threonine Kinases, Thiazolidinediones pharmacology, Triglycerides metabolism, Energy Metabolism drug effects, Glucose metabolism, Insulin metabolism, Insulin-Like Growth Factor I metabolism, Muscle Fibers, Skeletal metabolism, Oleic Acid metabolism, Oncorhynchus mykiss metabolism
- Abstract
The effects of insulin and IGF-I on fatty acid (FA) and glucose metabolism were examined using oleic acid or glucose as tracers in differentiated rainbow trout (Oncorhynchus mykiss) myotubes. Insulin and IGF-I significantly reduced the production of CO(2) from oleic acid with respect to the control values. IGF-I also significantly reduced the production of acid-soluble products (ASP) and the concentration of FA in the medium, while cellular triacylglycerols (TAG) tended to increase. Only insulin produced a significant accumulation of glycogen inside the cells in glucose distribution experiments. Incubation with catecholamines did not affect oleic acid metabolism. Cells treated with rapamycin [a target of rapamycin (TOR) inhibitor] significantly increased the oxidation of oleic acid to CO(2) and ASP, while the accumulation of TAG diminished. Rosiglitazone (a peroxisome proliferator-activated receptor gamma agonist) and etomoxir (a CPT-1 inhibitor) produced a severe and significant reduction in the production of CO(2) and ASP. Rosiglitazone and etomoxir also produced a significant accumulation of FA outside and inside the cells, respectively. No significant effects of these drugs on glucose distribution were observed. These data indicate that insulin and IGF-I act as anabolic hormones in trout myotubes in both oleic acid and glucose metabolism, although glucose oxidation appears to be less sensitive than FA oxidation to insulin and IGF-I. The use of rapamycin, etomoxir, and rosiglitazone may help us to understand the mechanisms of regulation of lipid metabolism in fish.
- Published
- 2010
- Full Text
- View/download PDF
35. Targets for TNFalpha-induced lipolysis in gilthead sea bream (Sparus aurata L.) adipocytes isolated from lean and fat juvenile fish.
- Author
-
Cruz-Garcia L, Saera-Vila A, Navarro I, Calduch-Giner J, and Pérez-Sánchez J
- Subjects
- Animals, Body Composition, Body Size, Fish Proteins genetics, Fish Proteins metabolism, Lipoprotein Lipase genetics, Lipoprotein Lipase metabolism, Sea Bream genetics, Sterol Esterase genetics, Sterol Esterase metabolism, Adipocytes metabolism, Lipolysis, Sea Bream metabolism, Tumor Necrosis Factor-alpha metabolism
- Abstract
The present study aimed to analyze adiposity heterogeneity and the role of liver X receptor (LXRalpha) and peroxisome proliferator-activated receptors (PPARs) as targets of tumour necrosis factor-alpha (TNFalpha) in gilthead sea bream (Sparus aurata L.). The screening of 20 fish at the beginning of the warm season identified two major groups with fat and lean phenotypes. Fat fish showed increased liver and mesenteric fat depots. This increased adiposity was concurrent in the adipose tissue to enhanced expression of lipoprotein lipase (LPL) whereas mRNA levels of the hormone-sensitive lipase (HSL) remained almost unchanged. The resulting LPL/HSL ratio was thereby highest in fat fish, which suggests that this group of fish has not reached its peak fat storage capacity. This is not surprising given the increased expression of PPARgamma in the absence of a counter-regulatory raise of TNFalpha. However, this lipolytic cytokine exerted dual effects in primary adipocyte cultures that differ within and between lean and fat fish. One set of fat fish did not respond to TNFalpha treatment whereas a second set exhibited a lipolytic response (increased glycerol release) that was apparently mediated by the downregulated expression of PPARbeta. In lean fish, TNFalpha exerted a strong and non-transcriptionally mediated lipolytic action. Alternatively, TNFalpha would inhibit lipid deposition via the downregulated expression of adipogenic nuclear factors (PPARgamma and LXRalpha). TNFalpha targets are therefore different in fish with lean and fat phenotypes, which is indicative of the complex network involved in the regulation of fish lipid metabolism.
- Published
- 2009
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.