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...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2022-02-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/75132 |
| _version_ | 1811180343182491648 |
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| author | Biliana Marcheva Benjamin J Weidemann Akihiko Taguchi Mark Perelis Kathryn Moynihan Ramsey Marsha V Newman Yumiko Kobayashi Chiaki Omura Jocelyn E Manning Fox Haopeng Lin Patrick E Macdonald Joseph Bass |
| author_facet | Biliana Marcheva Benjamin J Weidemann Akihiko Taguchi Mark Perelis Kathryn Moynihan Ramsey Marsha V Newman Yumiko Kobayashi Chiaki Omura Jocelyn E Manning Fox Haopeng Lin Patrick E Macdonald Joseph Bass |
| author_sort | Biliana Marcheva |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-04-11T09:01:38Z |
| format | Article |
| id | doaj.art-4ba877e38361469da7c8d4a16351a710 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-11T09:01:38Z |
| publishDate | 2022-02-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-4ba877e38361469da7c8d4a16351a7102022-12-22T04:32:46ZengeLife Sciences Publications LtdeLife2050-084X2022-02-011110.7554/eLife.75132P2Y1 purinergic receptor identified as a diabetes target in a small-molecule screen to reverse circadian β-cell failureBiliana Marcheva0https://orcid.org/0000-0001-8697-2625Benjamin J Weidemann1https://orcid.org/0000-0002-3747-2744Akihiko Taguchi2Mark Perelis3Kathryn Moynihan Ramsey4https://orcid.org/0000-0002-0691-7835Marsha V Newman5Yumiko Kobayashi6Chiaki Omura7Jocelyn E Manning Fox8Haopeng Lin9Patrick E Macdonald10Joseph Bass11https://orcid.org/0000-0002-1602-8601Department of Medicine, Division of Endocrinology, Metabolism and Molecular Medicine, Northwestern University Feinberg School of Medicine, Chicago, United StatesDepartment of Medicine, Division of Endocrinology, Metabolism and Molecular Medicine, Northwestern University Feinberg School of Medicine, Chicago, United StatesDepartment of Medicine, Division of Endocrinology, Metabolism and Molecular Medicine, Northwestern University Feinberg School of Medicine, Chicago, United States; Division of Endocrinology, Metabolism, Hematological Science and Therapeutics, Department of Bio-Signal Analysis, Yamaguchi University, Graduate School of Medicine, 1-1-1, Yamaguchi, JapanDepartment of Medicine, Division of Endocrinology, Metabolism and Molecular Medicine, Northwestern University Feinberg School of Medicine, Chicago, United States; Ionis Pharmaceuticals, Inc, Carlsbad, United StatesDepartment of Medicine, Division of Endocrinology, Metabolism and Molecular Medicine, Northwestern University Feinberg School of Medicine, Chicago, United StatesDepartment of Medicine, Division of Endocrinology, Metabolism and Molecular Medicine, Northwestern University Feinberg School of Medicine, Chicago, United StatesDepartment of Medicine, Division of Endocrinology, Metabolism and Molecular Medicine, Northwestern University Feinberg School of Medicine, Chicago, United StatesDepartment of Medicine, Division of Endocrinology, Metabolism and Molecular Medicine, Northwestern University Feinberg School of Medicine, Chicago, United StatesDepartment of Pharmacology, Alberta Diabetes Institute, University of Alberta, Edmonton, AB, CanadaDepartment of Pharmacology, Alberta Diabetes Institute, University of Alberta, Edmonton, AB, CanadaDepartment of Pharmacology, Alberta Diabetes Institute, University of Alberta, Edmonton, AB, CanadaDepartment of Medicine, Division of Endocrinology, Metabolism and Molecular Medicine, Northwestern University Feinberg School of Medicine, Chicago, United StatesThe 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.https://elifesciences.org/articles/75132circadian clockhigh-throughput screendiabetesivermectinpurinergic receptorinsulin |
| spellingShingle | Biliana Marcheva Benjamin J Weidemann Akihiko Taguchi Mark Perelis Kathryn Moynihan Ramsey Marsha V Newman Yumiko Kobayashi Chiaki Omura Jocelyn E Manning Fox Haopeng Lin Patrick E Macdonald Joseph Bass P2Y1 purinergic receptor identified as a diabetes target in a small-molecule screen to reverse circadian β-cell failure eLife circadian clock high-throughput screen diabetes ivermectin purinergic receptor insulin |
| 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 | circadian clock high-throughput screen diabetes ivermectin purinergic receptor insulin |
| url | https://elifesciences.org/articles/75132 |
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