CARD9 mediates glucose-stimulated insulin secretion in pancreatic beta cells

Abstract

Caspase recruitment domain containing protein 9 (CARD9) plays key regulatory role(s) in innate and adaptive immune responses. Recent evidence implicates CARD9 in the onset of metabolic diseases including insulin resistance. However, potential contributory roles of CARD9 in glucose-stimulated insulin secretion (GSIS) remain unknown. Herein, we report that CARD9 is expressed in human islets, rat islets, mouse islets and clonal INS-1 832/13 cells. Subcellularly, CARD9 is predominantly cytosolic (~75%) in INS-1 832/13 cells. siRNA-mediated depletion of CARD9 expression significantly (~50%) suppressed GSIS in INS-1 832/13 cells. Interestingly, glucose-induced activation of Rac1, a small G-protein, which is a requisite for GSIS to occur, is unaffected in CARD9-si transfected cells, suggesting that CARD9-mediates GSIS in a Rac1-independent fashion. Furthermore, insulin secretion promoted by KCl or mastoparan (a global G protein activator), remained resistant to CARD9 depletion in INS-1 832/13 cells. In addition, pharmacological inhibition (BRD5529) of interaction between CARD9 and TRIM62, its ubiquitin ligase, exerted no significant effects on GSIS. Lastly, depletion of CARD9 prevented glucose-induced p38, not ERK1/2 phosphorylation in beta cells. Based on these observations, we propose that CARD9 might regulate GSIS via a Rac1-independent and p38-dependent signaling module.