High-throughput screen reveals purinergic receptor as a therapeutic target in circadian ß-cell failure

We report a screen of 2,640 pharmacologically-active compounds in clock-deficient pancreatic ß cells and reveal a mechanism by which ivermectin induces insulin secretion and regulates transcription in wild type and P2ry1 receptor knockout BetaTC6 ß-cell lines by mRNA-seq. Overall design: Examination of mRNA expression changes in wild type and P2ry1 receptor knockout BetaTC6 ß-cell lines treated with 20 mM glucose in the presence or absence of ivermectin. The records have been updated with third-party re-analysis data as described below, on Jan 11, 2022: Sequencing data from the study under SRA accession ERP017126 (Bioproject: PRJEB15401; ENA-SUBMISSION: ERA705448) (PMID: 27667667) was downloaded and converted to fastq files using the commands 'prefetch' followed by 'fastq-dump' via sra-toolkit (v2.10.5). Each individual cell/file was aligned and transcript abundance quantified using RSEM (v1.3.3) with Hg38 (GRCh38.p12) as a reference. Raw single-cell expression count values were imported into Rstudio and count files were aggregated into count matrices for each condition (ctrl.data.txt, t2d.data.txt) for analysis using Seurat. Following low-quality cell removal using the author-provided metadata (https://www.ebi.ac.uk/arrayexpress/files/E-MTAB-5061/E-MTAB-5061.sdrf.txt), normalized expression values were used in uniform manifold approximation and projection (UMAP) dimensional reduction analyses to cluster distinct cell types (As outlined in E-MTAB-5061_analysis.R). E-MTAB-5061.sdrf.txt: Direct downloaded file from original authors including metadata on single-cell RNA-sequencing data. Available through https://www.ebi.ac.uk/arrayexpress/files/E-MTAB-5061/E-MTAB-5061.sdrf.txt ctrl.data.txt: Single-cell RNA count matrix for cells from non-type 2 diabetic donors t2d.data.txt: Single-cell RNA count matrix for cells from type 2 diabetic donors E-MTAB-5061_analysis.R: Custom script for Seurat analysis of single-cell RNA count data from the study E-MTAB-5061