| First Author | Marcheva B | Year | 2022 |
| Journal | Elife | Volume | 11 |
| PubMed ID | 35188462 | Mgi Jnum | J:343962 |
| Mgi Id | MGI:6885179 | Doi | 10.7554/eLife.75132 |
| Citation | Marcheva B, et al. (2022) P2Y1 purinergic receptor identified as a diabetes target in a small-molecule screen to reverse circadian beta-cell failure. Elife 11:e75132 |
| abstractText | 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 beta-cell failure. Our approach was to generate beta-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 beta-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. |
