哺乳动物VPS13D在内质网-线粒体接触位点维持中的功能研究

Interfaces between endoplasmic reticulum (ER) and mitochondria are emerging as critical platforms for many biological responses in eukaryotic cells, and changes in the extent of ER-mitochondrial contacts (EMCs) are reported in various models of neurodegenerative diseases including Alzheimer’s disease (AD), Parkinson’s disease (PD) and Amyotrophic lateral sclerosis(ALS). However, it is still unclear how the structure and functions of EMCs are maintained and regulated in mammalian cells. Using cutting-edge techniques including Crispr-cas9 gene editing and high-resolution live cell microscopy along with high-throughput siRNA library screening, we identified a novel gene VPS13D (Vacuolar protein sorting 13 homolog D) as a key player in maintaining the structure and functions of EMCs. The aim of this study is to identify molecules interacting with VPS13D, and further illustrate the molecular mechanisms how VPS13D functions in EMCs.

内质网和线粒体特化膜结构形成的接触位点 (ER-mitochondria contacts，EMCs) 被证实是一系列细胞核心生命过程的调控中枢，与主要的神经退行性疾病的发生发展直接相关。然而，目前对EMCs形态发生、维持和功能的调控机制尚知之甚少。本项目综合利用Crispr-cas9基因编辑、高分辨率活细胞成像等国际细胞生物学领域前沿技术,结合高通量siRNA文库筛选，以人骨肉瘤U2OS细胞系为模型，筛选出参与维系EMCs结构与功能稳定的关键新基因VPS13D（Vascular protein sorting protein 13D），并将进一步解析与VPS13D相互作用的效应分子，阐明VPS13D的作用机制。

内质网和线粒体特化膜结构形成的膜接触位点(ER-mitochondria membrane contacts, EMCs) 被证实是一系列细胞核心生命过程的调控中枢，与主要的神经退行疾病的发生发展直接相关。然而，目前对EMCs形态发生、维持和功能的调控机制尚知之甚少。本项目通过siRNA文库筛选，发现参与EMCs结构和功能调控的关键新分子VPS13D (Vascular protein sorting-associated protein 13D), 并解析VPS13D的关键相互作用分子(TSG101和VCP/P97)，阐明VPS13D调控细胞器相互作用的分子机制。相关研究成果以研究论文的形式发表在Nat. Comm. (第一标注)，J Cell Sci. (第一标注)和Molecular Biology of the Cell (第一标注)。VPS13D功能缺失导致的遗传性痉挛性共济失调(HSP)，因此我们的研究为这种疾病的诊断和治疗提供了理论依据和潜在新靶点。