Filippini Francesca, Nola Sébastien, Zahraoui Ahmed, Roger Kevin, Esmaili Mansoore, Sun Ji, Wojnacki José, Vlieghe Anaïs, Bun Philippe, Blanchon Stéphanie, Rain Jean-Christophe, Taymans Jean-Marc, Chartier-Harlin Marie-Christine, Guerrera Chiara, Galli Thierry
Université Paris Cité, Institute of Psychiatry and Neuroscience of Paris, INSERM U1266, Membrane Traffic in Healthy & Diseased Brain, 75014 Paris, France.
Université Paris Cité, Proteomics Platform Necker, Structure Fédérative de Recherche Necker, INSERM US24/CNRS UMS3633, 75015 Paris, France.
Cell Rep. 2023 Mar 28;42(3):112221. doi: 10.1016/j.celrep.2023.112221. Epub 2023 Mar 10.
The neuropeptide VGF was recently proposed as a neurodegeneration biomarker. The Parkinson's disease-related protein leucine-rich repeat kinase 2 (LRRK2) regulates endolysosomal dynamics, a process that involves SNARE-mediated membrane fusion and could regulate secretion. Here we investigate potential biochemical and functional links between LRRK2 and v-SNAREs. We find that LRRK2 directly interacts with the v-SNAREs VAMP4 and VAMP7. Secretomics reveals VGF secretory defects in VAMP4 and VAMP7 knockout (KO) neuronal cells. In contrast, VAMP2 KO "regulated secretion-null" and ATG5 KO "autophagy-null" cells release more VGF. VGF is partially associated with extracellular vesicles and LAMP1+ endolysosomes. LRRK2 expression increases VGF perinuclear localization and impairs its secretion. Retention using selective hooks (RUSH) assays show that a pool of VGF traffics through VAMP4+ and VAMP7+ compartments, and LRRK2 expression delays its transport to the cell periphery. Overexpression of LRRK2 or VAMP7-longin domain impairs VGF peripheral localization in primary cultured neurons. Altogether, our results suggest that LRRK2 might regulate VGF secretion via interaction with VAMP4 and VAMP7.
神经肽VGF最近被提议作为一种神经退行性变生物标志物。帕金森病相关蛋白富含亮氨酸重复激酶2（LRRK2）调节内溶酶体动力学，这一过程涉及SNARE介导的膜融合，并且可能调节分泌。在此，我们研究LRRK2与v-SNARE之间潜在的生化和功能联系。我们发现LRRK2直接与v-SNAREs VAMP4和VAMP7相互作用。分泌组学揭示了VAMP4和VAMP7基因敲除（KO）神经元细胞中VGF的分泌缺陷。相比之下，VAMP2基因敲除的“调节性分泌缺失”细胞和ATG5基因敲除的“自噬缺失”细胞释放更多的VGF。VGF部分与细胞外囊泡和LAMP1+内溶酶体相关。LRRK2的表达增加VGF在核周的定位并损害其分泌。使用选择性钩子滞留（RUSH）试验表明，一部分VGF通过VAMP4+和VAMP7+区室运输，并且LRRK2的表达延迟其向细胞周边的运输。LRRK2或VAMP7-长同源结构域的过表达损害原代培养神经元中VGF的周边定位。总之，我们的结果表明，LRRK2可能通过与VAMP4和VAMP7相互作用来调节VGF的分泌。
