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P2Y1 purinergic receptor identified as a diabetes target in a small-molecule screen to reverse circadian β-cell failure

The mammalian circadian clock drives daily oscillations in physiology and behavior through an autoregulatory transcription feedback loop present in central and peripheral cells. Ablation of the core clock within the endocrine pancreas of adult animals impairs the transcription and splicing of genes...

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Autores principales: Marcheva, Biliana, Weidemann, Benjamin J, Taguchi, Akihiko, Perelis, Mark, Ramsey, Kathryn Moynihan, Newman, Marsha V, Kobayashi, Yumiko, Omura, Chiaki, Manning Fox, Jocelyn E, Lin, Haopeng, Macdonald, Patrick E, Bass, Joseph
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8860442/
https://www.ncbi.nlm.nih.gov/pubmed/35188462
http://dx.doi.org/10.7554/eLife.75132
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author Marcheva, Biliana
Weidemann, Benjamin J
Taguchi, Akihiko
Perelis, Mark
Ramsey, Kathryn Moynihan
Newman, Marsha V
Kobayashi, Yumiko
Omura, Chiaki
Manning Fox, Jocelyn E
Lin, Haopeng
Macdonald, Patrick E
Bass, Joseph
author_facet Marcheva, Biliana
Weidemann, Benjamin J
Taguchi, Akihiko
Perelis, Mark
Ramsey, Kathryn Moynihan
Newman, Marsha V
Kobayashi, Yumiko
Omura, Chiaki
Manning Fox, Jocelyn E
Lin, Haopeng
Macdonald, Patrick E
Bass, Joseph
author_sort Marcheva, Biliana
collection PubMed
description The mammalian circadian clock drives daily oscillations in physiology and behavior through an autoregulatory transcription feedback loop present in central and peripheral cells. Ablation of the core clock within the endocrine pancreas of adult animals impairs the transcription and splicing of genes involved in hormone exocytosis and causes hypoinsulinemic diabetes. Here, we developed a genetically sensitized small-molecule screen to identify druggable proteins and mechanistic pathways involved in circadian β-cell failure. Our approach was to generate β-cells expressing a nanoluciferase reporter within the proinsulin polypeptide to screen 2640 pharmacologically active compounds and identify insulinotropic molecules that bypass the secretory defect in CRISPR-Cas9-targeted clock mutant β-cells. We validated hit compounds in primary mouse islets and identified known modulators of ligand-gated ion channels and G-protein-coupled receptors, including the antihelmintic ivermectin. Single-cell electrophysiology in circadian mutant mouse and human cadaveric islets revealed ivermectin as a glucose-dependent secretagogue. Genetic, genomic, and pharmacological analyses established the P2Y1 receptor as a clock-controlled mediator of the insulinotropic activity of ivermectin. These findings identify the P2Y1 purinergic receptor as a diabetes target based upon a genetically sensitized phenotypic screen.
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spelling pubmed-88604422022-02-23 P2Y1 purinergic receptor identified as a diabetes target in a small-molecule screen to reverse circadian β-cell failure Marcheva, Biliana Weidemann, Benjamin J Taguchi, Akihiko Perelis, Mark Ramsey, Kathryn Moynihan Newman, Marsha V Kobayashi, Yumiko Omura, Chiaki Manning Fox, Jocelyn E Lin, Haopeng Macdonald, Patrick E Bass, Joseph eLife Cell Biology The mammalian circadian clock drives daily oscillations in physiology and behavior through an autoregulatory transcription feedback loop present in central and peripheral cells. Ablation of the core clock within the endocrine pancreas of adult animals impairs the transcription and splicing of genes involved in hormone exocytosis and causes hypoinsulinemic diabetes. Here, we developed a genetically sensitized small-molecule screen to identify druggable proteins and mechanistic pathways involved in circadian β-cell failure. Our approach was to generate β-cells expressing a nanoluciferase reporter within the proinsulin polypeptide to screen 2640 pharmacologically active compounds and identify insulinotropic molecules that bypass the secretory defect in CRISPR-Cas9-targeted clock mutant β-cells. We validated hit compounds in primary mouse islets and identified known modulators of ligand-gated ion channels and G-protein-coupled receptors, including the antihelmintic ivermectin. Single-cell electrophysiology in circadian mutant mouse and human cadaveric islets revealed ivermectin as a glucose-dependent secretagogue. Genetic, genomic, and pharmacological analyses established the P2Y1 receptor as a clock-controlled mediator of the insulinotropic activity of ivermectin. These findings identify the P2Y1 purinergic receptor as a diabetes target based upon a genetically sensitized phenotypic screen. eLife Sciences Publications, Ltd 2022-02-21 /pmc/articles/PMC8860442/ /pubmed/35188462 http://dx.doi.org/10.7554/eLife.75132 Text en © 2022, Marcheva et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Cell Biology
Marcheva, Biliana
Weidemann, Benjamin J
Taguchi, Akihiko
Perelis, Mark
Ramsey, Kathryn Moynihan
Newman, Marsha V
Kobayashi, Yumiko
Omura, Chiaki
Manning Fox, Jocelyn E
Lin, Haopeng
Macdonald, Patrick E
Bass, Joseph
P2Y1 purinergic receptor identified as a diabetes target in a small-molecule screen to reverse circadian β-cell failure
title P2Y1 purinergic receptor identified as a diabetes target in a small-molecule screen to reverse circadian β-cell failure
title_full P2Y1 purinergic receptor identified as a diabetes target in a small-molecule screen to reverse circadian β-cell failure
title_fullStr P2Y1 purinergic receptor identified as a diabetes target in a small-molecule screen to reverse circadian β-cell failure
title_full_unstemmed P2Y1 purinergic receptor identified as a diabetes target in a small-molecule screen to reverse circadian β-cell failure
title_short P2Y1 purinergic receptor identified as a diabetes target in a small-molecule screen to reverse circadian β-cell failure
title_sort p2y1 purinergic receptor identified as a diabetes target in a small-molecule screen to reverse circadian β-cell failure
topic Cell Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8860442/
https://www.ncbi.nlm.nih.gov/pubmed/35188462
http://dx.doi.org/10.7554/eLife.75132
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