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Expression and functional assessment of candidate type 2 diabetes susceptibility genes identify four new genes contributing to human insulin secretion

OBJECTIVES: Genome-wide association studies (GWAS) have identified >100 loci independently contributing to type 2 diabetes (T2D) risk. However, translational implications for precision medicine and for the development of novel treatments have been disappointing, due to poor knowledge of how these...

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Autores principales: Ndiaye, Fatou K., Ortalli, Ana, Canouil, Mickaël, Huyvaert, Marlène, Salazar-Cardozo, Clara, Lecoeur, Cécile, Verbanck, Marie, Pawlowski, Valérie, Boutry, Raphaël, Durand, Emmanuelle, Rabearivelo, Iandry, Sand, Olivier, Marselli, Lorella, Kerr-Conte, Julie, Chandra, Vikash, Scharfmann, Raphaël, Poulain-Godefroy, Odile, Marchetti, Piero, Pattou, François, Abderrahmani, Amar, Froguel, Philippe, Bonnefond, Amélie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5444093/
https://www.ncbi.nlm.nih.gov/pubmed/28580277
http://dx.doi.org/10.1016/j.molmet.2017.03.011
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author Ndiaye, Fatou K.
Ortalli, Ana
Canouil, Mickaël
Huyvaert, Marlène
Salazar-Cardozo, Clara
Lecoeur, Cécile
Verbanck, Marie
Pawlowski, Valérie
Boutry, Raphaël
Durand, Emmanuelle
Rabearivelo, Iandry
Sand, Olivier
Marselli, Lorella
Kerr-Conte, Julie
Chandra, Vikash
Scharfmann, Raphaël
Poulain-Godefroy, Odile
Marchetti, Piero
Pattou, François
Abderrahmani, Amar
Froguel, Philippe
Bonnefond, Amélie
author_facet Ndiaye, Fatou K.
Ortalli, Ana
Canouil, Mickaël
Huyvaert, Marlène
Salazar-Cardozo, Clara
Lecoeur, Cécile
Verbanck, Marie
Pawlowski, Valérie
Boutry, Raphaël
Durand, Emmanuelle
Rabearivelo, Iandry
Sand, Olivier
Marselli, Lorella
Kerr-Conte, Julie
Chandra, Vikash
Scharfmann, Raphaël
Poulain-Godefroy, Odile
Marchetti, Piero
Pattou, François
Abderrahmani, Amar
Froguel, Philippe
Bonnefond, Amélie
author_sort Ndiaye, Fatou K.
collection PubMed
description OBJECTIVES: Genome-wide association studies (GWAS) have identified >100 loci independently contributing to type 2 diabetes (T2D) risk. However, translational implications for precision medicine and for the development of novel treatments have been disappointing, due to poor knowledge of how these loci impact T2D pathophysiology. Here, we aimed to measure the expression of genes located nearby T2D associated signals and to assess their effect on insulin secretion from pancreatic beta cells. METHODS: The expression of 104 candidate T2D susceptibility genes was measured in a human multi-tissue panel, through PCR-free expression assay. The effects of the knockdown of beta-cell enriched genes were next investigated on insulin secretion from the human EndoC-βH1 beta-cell line. Finally, we performed RNA-sequencing (RNA-seq) so as to assess the pathways affected by the knockdown of the new genes impacting insulin secretion from EndoC-βH1, and we analyzed the expression of the new genes in mouse models with altered pancreatic beta-cell function. RESULTS: We found that the candidate T2D susceptibility genes' expression is significantly enriched in pancreatic beta cells obtained by laser capture microdissection or sorted by flow cytometry and in EndoC-βH1 cells, but not in insulin sensitive tissues. Furthermore, the knockdown of seven T2D-susceptibility genes (CDKN2A, GCK, HNF4A, KCNK16, SLC30A8, TBC1D4, and TCF19) with already known expression and/or function in beta cells changed insulin secretion, supporting our functional approach. We showed first evidence for a role in insulin secretion of four candidate T2D-susceptibility genes (PRC1, SRR, ZFAND3, and ZFAND6) with no previous knowledge of presence and function in beta cells. RNA-seq in EndoC-βH1 cells with decreased expression of PRC1, SRR, ZFAND6, or ZFAND3 identified specific gene networks related to T2D pathophysiology. Finally, a positive correlation between the expression of Ins2 and the expression of Prc1, Srr, Zfand6, and Zfand3 was found in mouse pancreatic islets with altered beta-cell function. CONCLUSIONS: This study showed the ability of post-GWAS functional studies to identify new genes and pathways involved in human pancreatic beta-cell function and in T2D pathophysiology.
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spelling pubmed-54440932017-06-02 Expression and functional assessment of candidate type 2 diabetes susceptibility genes identify four new genes contributing to human insulin secretion Ndiaye, Fatou K. Ortalli, Ana Canouil, Mickaël Huyvaert, Marlène Salazar-Cardozo, Clara Lecoeur, Cécile Verbanck, Marie Pawlowski, Valérie Boutry, Raphaël Durand, Emmanuelle Rabearivelo, Iandry Sand, Olivier Marselli, Lorella Kerr-Conte, Julie Chandra, Vikash Scharfmann, Raphaël Poulain-Godefroy, Odile Marchetti, Piero Pattou, François Abderrahmani, Amar Froguel, Philippe Bonnefond, Amélie Mol Metab Original Article OBJECTIVES: Genome-wide association studies (GWAS) have identified >100 loci independently contributing to type 2 diabetes (T2D) risk. However, translational implications for precision medicine and for the development of novel treatments have been disappointing, due to poor knowledge of how these loci impact T2D pathophysiology. Here, we aimed to measure the expression of genes located nearby T2D associated signals and to assess their effect on insulin secretion from pancreatic beta cells. METHODS: The expression of 104 candidate T2D susceptibility genes was measured in a human multi-tissue panel, through PCR-free expression assay. The effects of the knockdown of beta-cell enriched genes were next investigated on insulin secretion from the human EndoC-βH1 beta-cell line. Finally, we performed RNA-sequencing (RNA-seq) so as to assess the pathways affected by the knockdown of the new genes impacting insulin secretion from EndoC-βH1, and we analyzed the expression of the new genes in mouse models with altered pancreatic beta-cell function. RESULTS: We found that the candidate T2D susceptibility genes' expression is significantly enriched in pancreatic beta cells obtained by laser capture microdissection or sorted by flow cytometry and in EndoC-βH1 cells, but not in insulin sensitive tissues. Furthermore, the knockdown of seven T2D-susceptibility genes (CDKN2A, GCK, HNF4A, KCNK16, SLC30A8, TBC1D4, and TCF19) with already known expression and/or function in beta cells changed insulin secretion, supporting our functional approach. We showed first evidence for a role in insulin secretion of four candidate T2D-susceptibility genes (PRC1, SRR, ZFAND3, and ZFAND6) with no previous knowledge of presence and function in beta cells. RNA-seq in EndoC-βH1 cells with decreased expression of PRC1, SRR, ZFAND6, or ZFAND3 identified specific gene networks related to T2D pathophysiology. Finally, a positive correlation between the expression of Ins2 and the expression of Prc1, Srr, Zfand6, and Zfand3 was found in mouse pancreatic islets with altered beta-cell function. CONCLUSIONS: This study showed the ability of post-GWAS functional studies to identify new genes and pathways involved in human pancreatic beta-cell function and in T2D pathophysiology. Elsevier 2017-04-08 /pmc/articles/PMC5444093/ /pubmed/28580277 http://dx.doi.org/10.1016/j.molmet.2017.03.011 Text en © 2017 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Original Article
Ndiaye, Fatou K.
Ortalli, Ana
Canouil, Mickaël
Huyvaert, Marlène
Salazar-Cardozo, Clara
Lecoeur, Cécile
Verbanck, Marie
Pawlowski, Valérie
Boutry, Raphaël
Durand, Emmanuelle
Rabearivelo, Iandry
Sand, Olivier
Marselli, Lorella
Kerr-Conte, Julie
Chandra, Vikash
Scharfmann, Raphaël
Poulain-Godefroy, Odile
Marchetti, Piero
Pattou, François
Abderrahmani, Amar
Froguel, Philippe
Bonnefond, Amélie
Expression and functional assessment of candidate type 2 diabetes susceptibility genes identify four new genes contributing to human insulin secretion
title Expression and functional assessment of candidate type 2 diabetes susceptibility genes identify four new genes contributing to human insulin secretion
title_full Expression and functional assessment of candidate type 2 diabetes susceptibility genes identify four new genes contributing to human insulin secretion
title_fullStr Expression and functional assessment of candidate type 2 diabetes susceptibility genes identify four new genes contributing to human insulin secretion
title_full_unstemmed Expression and functional assessment of candidate type 2 diabetes susceptibility genes identify four new genes contributing to human insulin secretion
title_short Expression and functional assessment of candidate type 2 diabetes susceptibility genes identify four new genes contributing to human insulin secretion
title_sort expression and functional assessment of candidate type 2 diabetes susceptibility genes identify four new genes contributing to human insulin secretion
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5444093/
https://www.ncbi.nlm.nih.gov/pubmed/28580277
http://dx.doi.org/10.1016/j.molmet.2017.03.011
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