Abstract
<jats:p>Insulin production is a cardinal feature of pancreatic β cells. Studies in rodents show that β cells can switch between low and high insulin gene activity states and that elevated insulin production makes β cells more vulnerable to stresses associated with diabetes. In people, genetically elevated insulin production increases the risk of type 1 diabetes. Via effects on obesity, hyperinsulinemia contributes to the pathogenesis of type 2 diabetes. Here, we characterize β cells in low and high INS gene activity states sorted from primary human islets transduced with INS-GFP adenovirus and differentiated INS-EGFP knock-in embryonic stem cells (SCβ cells). We profile β cell function, protein synthesis, resilience to diabetes associated stress, single β cell transcriptomes and their co-activity networks, and purified β cell proteomes. We show that human β cells transition between distinct states. High INS cells have elevated maturity marker mRNAs and proteins, increased protein translation, are larger, but also more susceptible to cell death when exposed to diabetes-relevant stresses. We also catalogue thousands of differences in proteins in high INS stem cell-derived β cells compared directly with high INS primary β cells. Our study improves our understanding of the delicate balance between insulin production and β cell resilience and guides the engineering of better β cells.</jats:p>