UPTAKE, TRANSPORT, AND SPREAD OF PRIONS

朊病毒的摄取、运输和传播

• 批准号: 8078393
• 负责人: DAVID A HARRIS
• 金额: $ 38万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-15 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8078393
• 关键词: UPTAKE; TRANSPORT; SPREAD; PRIONS

Prion diseases are fatal neurodegenerative disorders of humans and animals. They result from conversion of PrPC, a normal membrane glycoprotein into PrPSc, a conformationally altered isoform that is infectious in the absence of nucleic acid. Most exogenously acquired prion diseases arise by exposure to the infectious agent outside of the central nervous system (CNS). For this reason, a major focus of research in the field has been to understand how prions gain access to the CNS from the periphery, and how they spread within the spinalcord and brain. To facilitate studies of prion trafficking, we propose to utilize PrP molecules fused to enhanced green fluorescent protein (EGFP). Such PrP-EGFP fusion proteins make it possible to analyze the cellular trafficking of PrPSc in living cells in real time, and to avoid the artifacts associated with conventional immunocytochemical detection of PrPSc. We previously created Tg(PrP-EGFP) mice in which EGFP was inserted adjacent to the glycolipid attachment site of PrP. This form of PrP-EGFP serves a highly specific ligand that binds to and labels intracellular and extracellular deposits of PrPSc in prion-infected animals. However, this particular fusion protein is not itself converted into PrPSc-EGFP, and so is subject to several experimental limitations. In this two-year project, we propose to design improved PrP-EFGP constructs that can be converted efficiently into PrPSc-EGFP. We will then use these fluorescent proteins to visualize the transport of PrPSc along the axons of living neurons to determine whether this movement is an intra-axonal, motor-driven process, or occurs via a “domino mechanism” on the axolemma. We expect that the proposed experiments will provide important insights into how prions spread along nerves,and suggest how this process can be manipulated as a therapeutic or prophylactic strategy.

朊病毒病是人类和动物致命的神经退行性疾病。它们是由正常的膜糖蛋白PrPC转化为PrPSc而产生的，PrPSc是一种构象改变的异构体，在病毒中具有传染性