细胞膜蛋白TRPC1介导HSV-1病毒感染宿主细胞的作用及机制研究

Herpes simplex virus type 1 (HSV-1) causes a variety of diseases in humans, but current treatments cannot eradicate the virus from the body. Infection of HSV-1 is initiated by the recognition and binding of entry receptors on host cell membrane by viral glycoproteins on the capsid, the process will activate cellular calcium signals. However, the type of cell receptors and its molecular mechanism during HSV-1 infection is unclear by now. Based on our preliminary studies, this proposal aims at exploring the role of cell transient receptor potential channel C1 (TRPC1) in the HSV-1 infection. Our objectives includes: (1) to investigate the calcium homeostasis and the trafficking of TRPC1 during HSV-1 infection; (2) to explore the binding site of glycoprotein D (gD) on TRPC1, as well as the mechanism of infection to interfere with the trafficking of TRPC1 (3) to elucidate function of gD-TRPC1 complex during HSV-1 infection and replication; (4) to explore whether TRPC1 mediates HSV-1 infection in vivo. This study will shed light on understanding the mechanism of HSV-1 infection, as well as provide information to anti-HSV-1 methods development.

单纯疱疹病毒I型（HSV-1）引起人类多种疾病，目前无特效疗法根治其感染和复发。HSV-1感染起始于病毒包膜表面糖蛋白与宿主细胞膜受体间识别和连接，细胞内钙离子信号通路参与该过程。但宿主细胞膜表面受体种类、受体介导感染的过程、钙信号通路激活机制尚不明确。在前期研究基础上，项目拟利用免疫共沉淀、全细胞和单通道膜片钳、目标基因改造等方法，探讨宿主细胞膜的瞬时感受器电位阳离子通道亚型C1（TRPC1）介导HSV-1感染的机制。具体包括①探讨HSV-1对宿主细胞钙离子稳态及TRPC1活性的影响；②阐明病毒包膜最重要的糖蛋白gD结合宿主细胞TRPC1的分子原理，及其激活TRPC1的机制；③阐明TRPC1介导感染相关钙信号、调控病毒感染复制的机制；④在TRPC1基因敲除小鼠中，解析TRPC1介导HSV-1感染的机制。研究将利于丰富对HSV-1的感染机理的认识，助于抗HSV-1新疗法的研发。

单纯疱疹病毒I型（HSV-1）引起人类多种疾病，目前无特效疗法根治其感染和复发。HSV-1感染起始于病毒包膜表面糖蛋白与宿主细胞膜受体间识别和连接，细胞内钙离子信号通路参与该过程。但宿主细胞膜表面受体种类、受体介导感染的过程、钙信号通路激活机制尚不明确。本项目在细胞、动物及临床样本水平，利用免疫共沉淀、全细胞和单通道膜片钳、目标基因改造等方法，探讨了宿主细胞膜的瞬时感受器电位阳离子通道亚型C1（TRPC1）介导HSV-1感染的机制。研究结果发现：①HSV-1感染过程中，引起了宿主细胞HEp-2的内钙释放，进而激发了内钙释放介导的钙内流（SOCE），该SOCE由宿主细胞TRPC1介导；②随即，HSV-1包膜糖蛋白gD结合宿主细胞TRPC1的第三个膜外区。TRPC1继而辅助了HSV-1进入细胞；③TRPC1-gD相互作用在动物及临床样本中都得以体现；④基于这些结果，我们开发了两种HSV-1感染抑制剂。我们的研究结果解析了TRPC1介导HSV-1感染的机制。研究将利于丰富对HSV-1的感染机理的认识，助于抗HSV-1新疗法的研发。