Clinical and Experimental Endocrinology, KU Leuven, Leuven, Belgium.
Department of Molecular Cell Biology and School for Nutrition and Translational Research in Metabolism NUTRIM, Maastricht University, Maastricht, The Netherlands.
Cell Death Dis. 2019 Apr 5;10(4):309. doi: 10.1038/s41419-019-1518-0.
The 78-kDa glucose-regulated protein (GRP78) is an ubiquitously expressed endoplasmic reticulum chaperone, with a central role in maintaining protein homeostasis. Recently, an alternative role for GRP78 under stress conditions has been proposed, with stress-induced extracellular secretion and translocation of GRP78 to the cell surface where it acts as a multifunctional signaling receptor. Here we demonstrate translocation of GRP78 to the surface of human EndoC-βH1 cells and primary human islets upon cytokine exposure, in analogy to observations in rodent INS-1E and MIN6 beta cell lines. We show that GRP78 is shuttled via the anterograde secretory pathway, through the Golgi complex and secretory granules, and identify the DNAJ homolog subfamily C member 3 (DNAJC3) as a GRP78-interacting protein that facilitates its membrane translocation. Evaluation of downstream signaling pathways, using N- and C-terminal anti-GRP78 blocking antibodies, demonstrates that both GRP78 signaling domains initiate pro-apoptotic signaling cascades in beta cells. Extracellular GRP78 itself is identified as a ligand for cell surface GRP78 (sGRP78), increasing caspase 3/7 activity and cell death upon binding, which is accompanied by enhanced Chop and Bax mRNA expression. These results suggest that inflammatory cytokines induce a self-destructive pro-apoptotic feedback loop through the secretion and membrane translocation of GRP78. This proapoptotic function distinguishes the role of sGRP78 in beta cells from its reported anti-apoptotic and proliferative role in cancer cells, opening the road for the use of compounds that block sGRP78 as potential beta cell-preserving therapies in type 1 diabetes.
78kDa 葡萄糖调节蛋白(GRP78)是一种广泛表达的内质网伴侣,在维持蛋白质内稳态方面起着核心作用。最近,有人提出了 GRP78 在应激条件下的另一种作用,即在应激诱导下,GRP78 会被分泌到细胞外,并转移到细胞表面,在细胞表面,它作为一种多功能信号受体发挥作用。在这里,我们证明了细胞因子暴露后,人 EndoC-βH1 细胞和原代人胰岛中 GRP78 会转移到细胞表面,这与在啮齿动物 INS-1E 和 MIN6 β细胞系中的观察结果类似。我们表明,GRP78 通过顺行分泌途径穿梭,穿过高尔基体复合体和分泌颗粒,并确定 DNAJ 同源物亚家族 C 成员 3(DNAJC3)是一种与 GRP78 相互作用的蛋白,可促进其膜转位。使用 N 端和 C 端抗 GRP78 阻断抗体评估下游信号通路表明,GRP78 的两个信号结构域都在 β细胞中启动促凋亡信号级联反应。细胞外的 GRP78 本身被鉴定为细胞表面 GRP78(sGRP78)的配体,在结合后增加 caspase 3/7 活性和细胞死亡,同时伴随着 Chop 和 Bax mRNA 表达的增强。这些结果表明,炎症细胞因子通过 GRP78 的分泌和膜转位诱导一种自毁性的促凋亡反馈环。这种促凋亡功能区分了 sGRP78 在 β细胞中的作用与其在癌细胞中报道的抗凋亡和增殖作用,为使用阻断 sGRP78 的化合物作为 1 型糖尿病中潜在的β细胞保护疗法开辟了道路。