Cargando…

High-Throughput Quantitative Screening of Glucose-Stimulated Insulin Secretion and Insulin Content Using Automated MALDI-TOF Mass Spectrometry

Type 2 diabetes (T2D) is a metabolic disorder characterized by loss of pancreatic β-cell function, decreased insulin secretion and increased insulin resistance, that affects more than 537 million people worldwide. Although several treatments are proposed to patients suffering from T2D, long-term con...

Descripción completa

Detalles Bibliográficos
Autores principales: Delannoy, Clément Philippe, Heuson, Egon, Herledan, Adrien, Oger, Frederik, Thiroux, Bryan, Chevalier, Mickaël, Gromada, Xavier, Rolland, Laure, Froguel, Philippe, Deprez, Benoit, Paul, Sébastien, Annicotte, Jean-Sébastien
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10047017/
https://www.ncbi.nlm.nih.gov/pubmed/36980190
http://dx.doi.org/10.3390/cells12060849
_version_ 1785013816528994304
author Delannoy, Clément Philippe
Heuson, Egon
Herledan, Adrien
Oger, Frederik
Thiroux, Bryan
Chevalier, Mickaël
Gromada, Xavier
Rolland, Laure
Froguel, Philippe
Deprez, Benoit
Paul, Sébastien
Annicotte, Jean-Sébastien
author_facet Delannoy, Clément Philippe
Heuson, Egon
Herledan, Adrien
Oger, Frederik
Thiroux, Bryan
Chevalier, Mickaël
Gromada, Xavier
Rolland, Laure
Froguel, Philippe
Deprez, Benoit
Paul, Sébastien
Annicotte, Jean-Sébastien
author_sort Delannoy, Clément Philippe
collection PubMed
description Type 2 diabetes (T2D) is a metabolic disorder characterized by loss of pancreatic β-cell function, decreased insulin secretion and increased insulin resistance, that affects more than 537 million people worldwide. Although several treatments are proposed to patients suffering from T2D, long-term control of glycemia remains a challenge. Therefore, identifying new potential drugs and targets that positively affect β-cell function and insulin secretion remains crucial. Here, we developed an automated approach to allow the identification of new compounds or genes potentially involved in β-cell function in a 384-well plate format, using the murine β-cell model Min6. By using MALDI-TOF mass spectrometry, we implemented a high-throughput screening (HTS) strategy based on the automation of a cellular assay allowing the detection of insulin secretion in response to glucose, i.e., the quantitative detection of insulin, in a miniaturized system. As a proof of concept, we screened siRNA targeting well-know β-cell genes and 1600 chemical compounds and identified several molecules as potential regulators of insulin secretion and/or synthesis, demonstrating that our approach allows HTS of insulin secretion in vitro.
format Online
Article
Text
id pubmed-10047017
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-100470172023-03-29 High-Throughput Quantitative Screening of Glucose-Stimulated Insulin Secretion and Insulin Content Using Automated MALDI-TOF Mass Spectrometry Delannoy, Clément Philippe Heuson, Egon Herledan, Adrien Oger, Frederik Thiroux, Bryan Chevalier, Mickaël Gromada, Xavier Rolland, Laure Froguel, Philippe Deprez, Benoit Paul, Sébastien Annicotte, Jean-Sébastien Cells Article Type 2 diabetes (T2D) is a metabolic disorder characterized by loss of pancreatic β-cell function, decreased insulin secretion and increased insulin resistance, that affects more than 537 million people worldwide. Although several treatments are proposed to patients suffering from T2D, long-term control of glycemia remains a challenge. Therefore, identifying new potential drugs and targets that positively affect β-cell function and insulin secretion remains crucial. Here, we developed an automated approach to allow the identification of new compounds or genes potentially involved in β-cell function in a 384-well plate format, using the murine β-cell model Min6. By using MALDI-TOF mass spectrometry, we implemented a high-throughput screening (HTS) strategy based on the automation of a cellular assay allowing the detection of insulin secretion in response to glucose, i.e., the quantitative detection of insulin, in a miniaturized system. As a proof of concept, we screened siRNA targeting well-know β-cell genes and 1600 chemical compounds and identified several molecules as potential regulators of insulin secretion and/or synthesis, demonstrating that our approach allows HTS of insulin secretion in vitro. MDPI 2023-03-09 /pmc/articles/PMC10047017/ /pubmed/36980190 http://dx.doi.org/10.3390/cells12060849 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Delannoy, Clément Philippe
Heuson, Egon
Herledan, Adrien
Oger, Frederik
Thiroux, Bryan
Chevalier, Mickaël
Gromada, Xavier
Rolland, Laure
Froguel, Philippe
Deprez, Benoit
Paul, Sébastien
Annicotte, Jean-Sébastien
High-Throughput Quantitative Screening of Glucose-Stimulated Insulin Secretion and Insulin Content Using Automated MALDI-TOF Mass Spectrometry
title High-Throughput Quantitative Screening of Glucose-Stimulated Insulin Secretion and Insulin Content Using Automated MALDI-TOF Mass Spectrometry
title_full High-Throughput Quantitative Screening of Glucose-Stimulated Insulin Secretion and Insulin Content Using Automated MALDI-TOF Mass Spectrometry
title_fullStr High-Throughput Quantitative Screening of Glucose-Stimulated Insulin Secretion and Insulin Content Using Automated MALDI-TOF Mass Spectrometry
title_full_unstemmed High-Throughput Quantitative Screening of Glucose-Stimulated Insulin Secretion and Insulin Content Using Automated MALDI-TOF Mass Spectrometry
title_short High-Throughput Quantitative Screening of Glucose-Stimulated Insulin Secretion and Insulin Content Using Automated MALDI-TOF Mass Spectrometry
title_sort high-throughput quantitative screening of glucose-stimulated insulin secretion and insulin content using automated maldi-tof mass spectrometry
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10047017/
https://www.ncbi.nlm.nih.gov/pubmed/36980190
http://dx.doi.org/10.3390/cells12060849
work_keys_str_mv AT delannoyclementphilippe highthroughputquantitativescreeningofglucosestimulatedinsulinsecretionandinsulincontentusingautomatedmalditofmassspectrometry
AT heusonegon highthroughputquantitativescreeningofglucosestimulatedinsulinsecretionandinsulincontentusingautomatedmalditofmassspectrometry
AT herledanadrien highthroughputquantitativescreeningofglucosestimulatedinsulinsecretionandinsulincontentusingautomatedmalditofmassspectrometry
AT ogerfrederik highthroughputquantitativescreeningofglucosestimulatedinsulinsecretionandinsulincontentusingautomatedmalditofmassspectrometry
AT thirouxbryan highthroughputquantitativescreeningofglucosestimulatedinsulinsecretionandinsulincontentusingautomatedmalditofmassspectrometry
AT chevaliermickael highthroughputquantitativescreeningofglucosestimulatedinsulinsecretionandinsulincontentusingautomatedmalditofmassspectrometry
AT gromadaxavier highthroughputquantitativescreeningofglucosestimulatedinsulinsecretionandinsulincontentusingautomatedmalditofmassspectrometry
AT rollandlaure highthroughputquantitativescreeningofglucosestimulatedinsulinsecretionandinsulincontentusingautomatedmalditofmassspectrometry
AT froguelphilippe highthroughputquantitativescreeningofglucosestimulatedinsulinsecretionandinsulincontentusingautomatedmalditofmassspectrometry
AT deprezbenoit highthroughputquantitativescreeningofglucosestimulatedinsulinsecretionandinsulincontentusingautomatedmalditofmassspectrometry
AT paulsebastien highthroughputquantitativescreeningofglucosestimulatedinsulinsecretionandinsulincontentusingautomatedmalditofmassspectrometry
AT annicottejeansebastien highthroughputquantitativescreeningofglucosestimulatedinsulinsecretionandinsulincontentusingautomatedmalditofmassspectrometry