trem2突变加剧神经退行性疾病发生风险的作用机制研究微生物学专业论文.docxVIP

摘要TREM2的胞外段从而作为TREM2的一个新配体。我们细致地探讨了apoE与 摘要 TREM2的胞外段从而作为TREM2的一个新配体。我们细致地探讨了apoE与 TREM2结合的特点。通过表面固相结合实验，发现apoE3(10nM)与TREM2 结合的解离常数为150．8nM。同时我们发现，R47H、R62H、Y38C、T66M突变 形式的TREM2与apoE3的亲和性显著降低。通过对胞外不同长度的TREM2分 析，我们证实了apoE主要结合在TREM2 1．71氨基端序列上。通过分析该区段 氨基酸特点构建了TREM2胞外正电荷氨基酸突变成丙氨酸的突变(K42A、R46A、 R47A、K48A、R52A、K57A)，发现正电荷氨基酸突变的sTREM2与apoE3的 亲和力降低， 说明apoE与TREM2的结合主要通过静电相互作用，TREM2的 精氨酸和赖氨酸对于二者的结合起到关键作用。 本研究证实了疾病相关的TREM2突变影响了其在细胞中的转运及sTREM2 的分泌，并且发现这些突变影响了sTREM2在炎症和凋亡中的功能，我们证实 这些突变降低了其与配体apoE的亲和力。总之，通过对TREM2突变增加神经 退行性疾病发生风险的作用机制进行细致探讨，可以了解sTREM2如何参与神 经退行性疾病的发生发展及疾病进程，也可以为靶向sTREM2的神经退行性疾 病药物研发提供理论基础。 关键词：sTREM2；apoE；小胶质细胞；炎症；阿尔茨海默症 万方数据 AbstractAbstract Abstract Abstract Triggering receptor expressed myeliod cells 2(TREM2)is an innate immune receptor mainly expressed on the surface of microglia．TREM2 plays important roles in cellular inflammation and phagocytosis via TREM2／DAP 1 2 complex．TREM2 belongs to type-I transmembrane proteins with an ectodomain that is proteolytically cleaved and released into the extracellular space as soluble form(soluble TREM2， sTREM2)．It is well—known that mutations in TREM2 gene cause Nasu—Hakola Disease(NHD)．Recently,genome—wide association studies have shown that TREM2 mutations markedly increase the risk for Alzheimer’S disease(AD)．The most commonly associated variant，rencoding R47H)has a similar odds ratio to APOE4．Subsequent research shows that TREM2 variants increase the risk for spectrum of neurodegenerative disorders，including frontotempoml dementia(FTD)， Parkinson’S disease(PD)and amyotrophic lateral sclerosis(ALS)．Therefore TREM2 plays important roles in neurodegenerative disorders，thou曲the mechanism remains elusive．APOE is one of the strongest risk factors for AD that exists in three isofroms (￡2，￡3，￡4)．Among them，apoE4 is associated with increased prevalence of AD and lower age of onset． Herein，we firstly constructed plasmids carried the coding sequences for the extracellular domain of TREM2(sTREM2，1-1 7 1)and diseas