分析错误: Calcium-dependent transcriptional changes in human pancreatic islet cells reveal functional diversity in islet cell subtypes

Aims/hypothesis

Pancreatic islets depend on cytosolic calcium (Ca²⁺) to trigger the secretion of glucoregulatory hormones and trigger transcriptional regulation of genes important for islet response to stimuli. To date, there has not been an attempt to profile Ca²⁺-regulated gene expression in all islet cell types. Our aim was to construct a large single-cell transcriptomic dataset from human islets exposed to conditions that would acutely induce or inhibit intracellular Ca²⁺ signalling, while preserving biological heterogeneity.

Methods

We exposed intact human islets from three donors to the following conditions: (1) 2.8 mmol/l glucose; (2) 16 mmol/l glucose and 40 mmol/l KCl to maximally stimulate Ca²⁺ signalling; and (3) 16 mmol/l glucose, 40 mmol/l KCl and 5 mmol/l EGTA (Ca²⁺ chelator) to inhibit Ca²⁺signalling, for 1 h. We sequenced 68,650 cells from all islet cell types, and further subsetted the cells to form an endocrine cell-specific dataset of 59,373 cells expressing INS, GCG, SST or PPY. We compared transcriptomes across conditions to determine the differentially expressed Ca²⁺-regulated genes in each endocrine cell type, and in each endocrine cell subcluster of alpha and beta cells.

Results

Based on the number of Ca²⁺-regulated genes, we found that each alpha and beta cell cluster had a different magnitude of Ca²⁺ response. We also showed that polyhormonal clusters expressing both INS and GCG, or both INS and SST, are defined by Ca²⁺-regulated genes specific to each cluster. Finally, we identified the gene PCDH7 from the beta cell clusters that had the highest number of Ca²⁺-regulated genes, and showed that cells expressing cell surface PCDH7 protein have enhanced glucose-stimulated insulin secretory function.

Conclusions/interpretation

Here we use our large-scale, multi-condition, single-cell dataset to show that human islets have cell-type-specific Ca²⁺-regulated gene expression profiles, some of them specific to subpopulations. In our dataset, we identify PCDH7 as a novel marker of beta cells having an increased number of Ca²⁺-regulated genes and enhanced insulin secretory function.