Dual-Input Regulation of β-Cell Proliferation by ATF6α and Glucose via E2F1

Finding ways to increase β-cell mass is a key goal of diabetes research. During elevated insulin demand, β-cells turn on endoplasmic reticulum (ER) stress response pathways, and some β-cells enter the cell cycle. ER stress response protein activating transcription factor 6 (ATF6α) induces β-cell proliferation, but only in high glucose. The mechanism by which ATF6α increases proliferation, and the reasons for glucose dependence, remain unknown. Here we show that ATF6α activation in mouse and human islet cells increases expression of E2F1, a key cell cycle driver. E2F1 was required for ATF6α-induced proliferation in high glucose. However, E2F1 remained inactive in normal glucose, possibly because retinoblastoma (Rb), a direct E2F1 inhibitor, was in its dephosphorylated, active state. Indeed, inducing Rb phosphorylation by overexpressing cyclin-dependent kinase 4 (CDK4) allowed ATF6α to increase E2F1 activity and β-cell proliferation in normal glucose. E2F1 expression increased in an ATF6α-dependent manner during generalized ER stress by thapsigargin treatment. Importantly, in human β-cells, ATF6α failed to synergize with high glucose to induce proliferation, but the synergy was rescued by adding back CDK6. Taken together, this study establishes a new dual-input β-cell proliferation regulatory mechanism integrating ER load with current glycemic conditions via CDK4/6, in which Rb phosphorylation serves as a glucose sensor that permits ATF6α-driven proliferation. ARTICLE HIGHLIGHTS: Endoplasmic reticulum stress response mediator activating transcription factor 6 (ATF6α) increases pancreatic β-cell proliferation in a glucose-dependent manner, but the mechanism remains unknown. ATF6α activation upregulated mRNA and protein expression of E2F1, a key G1/S phase transition regulator; however, E2F1 activity only increased in high glucose. Glucose dependence of E2F1 activity was mediated by cyclin-dependent kinase 4/6 phosphorylation of retinoblastoma (Rb) protein, derepressing E2F1 in high glucose. Generalized endoplasmic reticulum stressor thapsigargin increased E2F1 abundance in an ATF6-dependent manner. ATF6α increased E2F1 expression in human β-cells and increased human β-cell proliferation when cyclin-dependent kinase 6 (CDK6) was coexpressed.