SYT11 and SYT13, two calcium‐insensitive synaptotagmins, are downregulated in islets from type 2 diabetic donors, but their function in insulin secretion is unknown. To address this, we investigated the physiological role of these two synaptotagmins in insulin‐secreting cells.

Correlations between gene expression levels were performed using previously described RNA‐seq data on islets from 188 human donors. SiRNA knockdown was performed in EndoC‐βH1 and INS‐1 832/13 cells. Insulin secretion was measured with ELISA. Patch‐clamp was used for single‐cell electrophysiology. Confocal microscopy was used to determine intracellular localization.

Human islet expression of the transcription factor PDX1 was positively correlated with SYT11 (p = 2.4e⁻¹⁰) and SYT13 (p < 2.2e⁻¹⁶). Syt11 and Syt13 both co‐localized with insulin, indicating their localization in insulin granules. Downregulation of Syt11 in INS‐1 832/13 cells (siSYT11) resulted in increased basal and glucose‐induced insulin secretion. Downregulation of Syt13 (siSYT13) decreased insulin secretion induced by glucose and K⁺. Interestingly, the cAMP‐raising agent forskolin was unable to enhance insulin secretion in siSYT13 cells. There was no difference in insulin content, exocytosis, or voltage‐gated Ca²⁺ currents in the two models. Double knockdown of Syt11 and Syt13 (DKD) resembled the results in siSYT13 cells.

SYT11 and SYT13 have similar localization and transcriptional regulation, but they regulate insulin secretion differentially. While downregulation of SYT11 might be a compensatory mechanism in type‐2 diabetes, downregulation of SYT13 reduces the insulin secretory response and overrules the compensatory regulation of SYT11 in a way that could aggravate the disease.