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1. The Combined Partial Knockdown of CBS and MPST Genes Induces Inflammation, Impairs Adipocyte Function-Related Gene Expression and Disrupts Protein Persulfidation in Human Adipocytes

Author

Jose Manuel Fernández-Real, Jèssica Latorre Luque, José María Moreno-Navarrete, Luis C. Romero, and Angeles Aroca

Subjects
Physiology, human adipocytes, adipogenesis, inflammation, protein persulfidation, Clinical Biochemistry, Cell Biology, Molecular Biology, Biochemistry
Abstract

Recent studies in mice and humans demonstrated the relevance of H2S synthesising enzymes, such as CTH, CBS, and MPST, in the physiology of adipose tissue and the differentiation of preadipocyte into adipocytes. Here, our objective was to investigate the combined role of CTH, CBS, and MPST in the preservation of adipocyte protein persulfidation and adipogenesis. Combined partial CTH, CBS, and MPST gene knockdown was achieved treating fully human adipocytes with siRNAs against these transcripts (siRNA_MIX). Adipocyte protein persulfidation was analyzed using label-free quantitative mass spectrometry coupled with a dimedone-switch method for protein labeling and purification. Proteomic analysis quantified 216 proteins with statistically different levels of persulfidation in KD cells compared to control adipocytes. In fully differentiated adipocytes, CBS and MPST mRNA and protein levels were abundant, while CTH expression was very low. It is noteworthy that siRNA_MIX administration resulted in a significant decrease in CBS and MPST expression, without impacting on CTH. The combined partial knockdown of the CBS and MPST genes resulted in reduced cellular sulfide levels in parallel to decreased expression of relevant genes for adipocyte biology, including adipogenesis, mitochondrial biogenesis, and lipogenesis, but increased proinflammatory- and senescence-related genes. It should be noted that the combined partial knockdown of CBS and MPST genes also led to a significant disruption in the persulfidation pattern of the adipocyte proteins. Although among the less persulfidated proteins, we identified several relevant proteins for adipocyte adipogenesis and function, among the most persulfidated, key mediators of adipocyte inflammation and dysfunction as well as some proteins that might play a positive role in adipogenesis were found. In conclusion, the current study indicates that the combined partial elimination of CBS and MPST (but not CTH) in adipocytes affects the expression of genes related to the maintenance of adipocyte function and promotes inflammation, possibly by altering the pattern of protein persulfidation in these cells, suggesting that these enzymes were required for the functional maintenance of adipocytes.

Published
2022
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2. Publisher Correction: Molecular phenomics and metagenomics of hepatic steatosis in non-diabetic obese women

Author

Julie Charpentier, Ottavia Porzio, Josep Puig, Matteo Serino, Iris Cardolini, Richard H. Barton, Fabienne Foufelle, Marina Cardellini, Fadila Benhamed, José Manuel Fernández-Real, Massimo Federici, Marc-Emmanuel Dumas, Mark Woodbridge, Jeremy K. Nicholson, Julien Chilloux, Antonis Myridakis, Wifredo Ricart, Vincent Blasco-Baque, Paolo Gentileschi, Christopher Tomlinson, Christophe Heymes, Francesca Davato, Elaine Holmes, José María Moreno-Navarrete, Sarah Butcher, Gemma Xifra, Jèssica Latorre Luque, Rémy Burcelin, Lesley Hoyles, Catherine Postic, Vincent Azalbert, Elodie Anthony, Laura Martinez-Gili, James Abbott, Frédéric Lopez, Imperial College London, University Hospital of Girona, University of Rome 'Tor Vergeta', Università degli Studi di Roma Tor Vergata [Roma], Institut de médecine moléculaire de Rangueil (I2MR), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées- Institut Fédératif de Recherche Bio-médicale Institution (IFR150)-Institut National de la Santé et de la Recherche Médicale (INSERM), Department of Embryology and Histopathology, Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Institut des Maladies Métaboliques et Cardiovasculaires (I2MC), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Cardiff Business School, Cardiff University, Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-IFR150-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Toulouse III - Paul Sabatier (UT3), and Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées

Subjects
0301 basic medicine, business.industry, Published Erratum, [SDV]Life Sciences [q-bio], MEDLINE, General Medicine, Bioinformatics, medicine.disease, General Biochemistry, Genetics and Molecular Biology, 3. Good health, 03 medical and health sciences, 030104 developmental biology, 0302 clinical medicine, Phenomics, Metagenomics, 030220 oncology & carcinogenesis, ComputingMethodologies_DOCUMENTANDTEXTPROCESSING, Medicine, Steatosis, business, GeneralLiterature_REFERENCE(e.g.,dictionaries,encyclopedias,glossaries), ComputingMilieux_MISCELLANEOUS, Non diabetic
Abstract

In the version of this article originally published, the received date was missing. It should have been listed as 2 January 2018. The error has been corrected in the HTML and PDF versions of this article.

Published
2018

3. Molecular phenomics and metagenomics of hepatic steatosis in non-diabetic obese women

Author

Jèssica Latorre Luque, Marina Cardellini, Marc-Emmanuel Dumas, Gemma Xifra, Francesca Davato, Josep Puig, Vincent Blasco-Baque, Iris Cardolini, Elodie Anthony, José Manuel Fernández-Real, Fadila Benhamed, Laura Martinez-Gili, Massimo Federici, Rémy Burcelin, Wifredo Ricart, Lesley Hoyles, Julie Charpentier, Catherine Postic, Richard H. Barton, Frédéric Lopez, José María Moreno-Navarrete, Sarah Butcher, Antonis Myridakis, Matteo Serino, James Abbott, Fabienne Foufelle, Vincent Azalbert, Paolo Gentileschi, Christopher Tomlinson, Julien Chilloux, Christophe Heymes, Mark Woodbridge, Elaine Holmes, Ottavia Porzio, Jeremy K. Nicholson, Commission of the European Communities, Medical Research Council (MRC), Imperial College London, Instituto de Salud Carlos III [Madrid] (ISC), University of Rome 'Tor Vergeta', Institut de médecine moléculaire de Rangueil (I2MR), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-IFR150-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut des Maladies Métaboliques et Cardiovasculaires (I2MC), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Cardiff Business School, Cardiff University, University of Rome TorVergata, Bambino Gesù Children’s Hospital [Rome, Italy], Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université de Paris (UP), Department of Embryology and Histopathology, Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Nutrition et cerveau (U1213, U855), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon, Section of Diabetes, Endocrinology and Nutrition, University Hospital of Girona-Biomedical Research Institute 'Dr Josep Trueta'-CIBERobn Fisiopatología de la Obesidad y Nutrición, Department d'enginyeria quimica agraria i tecnologia agroalimentaria, Universitat de Girona (UdG), Hosp Josep Trueta, Inst Invest Biomed Girona, Dept Diabet Endocrinol & Nutr, Institut Català de la Salut (ICS), Università degli Studi di Roma Tor Vergata [Roma], Centre de Recherches en Cancérologie de Toulouse (CRCT), Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Centre de Recherche des Cordeliers (CRC), Université Pierre et Marie Curie - Paris 6 (UPMC)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institute of Biotechnology, [Institut Cochin] Département Endocrinologie, métabolisme, diabète (EMD) (EMD), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Biomolecular Medicine, Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université Paris Diderot - Paris 7 (UPD7)-École pratique des hautes études (EPHE)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), and Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées- Institut Fédératif de Recherche Bio-médicale Institution (IFR150)-Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
0301 basic medicine, Settore MED/09 - Medicina Interna, IMPACT, [SDV]Life Sciences [q-bio], Physiology, Research & Experimental Medicine, PHENOTYPE, GLUCOSE, Transcriptome, Cohort Studies, Diabetes mellitus genetics, Mice, Non-alcoholic Fatty Liver Disease, Settore MED/49 - Scienze Tecniche Dietetiche Applicate, ComputingMilieux_MISCELLANEOUS, Cells, Cultured, 11 Medical and Health Sciences, INSULIN-RESISTANCE, Settore BIO/12, Microbiota, Fatty liver, Confounding Factors, Epidemiologic, General Medicine, ASSOCIATION, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, Fecal Microbiota Transplantation, 3. Good health, Medicine, Research & Experimental, CARDIOVASCULAR-DISEASE, Metabolome, Female, Life Sciences & Biomedicine, Biochemistry & Molecular Biology, HUMAN GUT MICROBIOME, Immunology, Biology, Phenome, METABOLISM, Article, General Biochemistry, Genetics and Molecular Biology, DIET, 03 medical and health sciences, [CHIM.ANAL]Chemical Sciences/Analytical chemistry, medicine, Diabetes Mellitus, [CHIM]Chemical Sciences, Animals, Humans, Metabolomics, Microbiome, Obesity, FATTY LIVER-DISEASE, Science & Technology, Cell Biology, medicine.disease, Confounding Factors (Epidemiology), Settore MED/18, Fatty Liver, Settore MED/18 - Chirurgia Generale, 030104 developmental biology, Metagenomics, Hepatocytes, [INFO.INFO-BI]Computer Science [cs]/Bioinformatics [q-bio.QM], Steatosis
Abstract

Hepatic steatosis is a multifactorial condition that is often observed in obese patients and is a prelude to non-alcoholic fatty liver disease. Here, we combine shotgun sequencing of fecal metagenomes with molecular phenomics (hepatic transcriptome and plasma and urine metabolomes) in two well-characterized cohorts of morbidly obese women recruited to the FLORINASH study. We reveal molecular networks linking the gut microbiome and the host phenome to hepatic steatosis. Patients with steatosis have low microbial gene richness and increased genetic potential for the processing of dietary lipids and endotoxin biosynthesis (notably from Proteobacteria), hepatic inflammation and dysregulation of aromatic and branched-chain amino acid metabolism. We demonstrated that fecal microbiota transplants and chronic treatment with phenylacetic acid, a microbial product of aromatic amino acid metabolism, successfully trigger steatosis and branched-chain amino acid metabolism. Molecular phenomic signatures were predictive (area under the curve = 87%) and consistent with the gut microbiome having an effect on the steatosis phenome (>75% shared variation) and, therefore, actionable via microbiome-based therapies. Metabolic activity of specific human gut microorganisms contributes to liver steatosis in obese women.

Published
2018

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