Biochemistry Of Scrapie Pathogenesis

传染性海绵状脑病(TSE或Prion病)是一种致命的神经退行性疾病，如瘙痒病、克雅氏病(CJD)、牛海绵状脑病和慢性衰弱病(CWD)。我们的项目旨在了解和阻止正常PrP(PrP-sen)到PrP-RES的转换，PrP-RES是一种异常形式的PrP蛋白(PrP)，似乎是TSE传播和发病的基础。我们采用了各种各样的细胞生物学、生化、生物物理和活体实验方法。 在过去的一年里，我们利用场流分离、光散射、电子显微镜和电泳法继续鉴定和分离TSE疾病最大和最小的感染单位，2)在体内外发现了作为强有力的PrP-res抑制剂和抗TSE药物的简并硫代寡核苷酸，3)表征了PrP-res侵袭神经元和神经胶质细胞、启动感染并沿着神经元突起运输到与其他细胞接触的肢体的过程，使用共聚焦显微镜，4)建立了第一个感染CWD的组织培养细胞系，并鉴定了第一个CWD复制抑制物，5)继续在动物身上测试卟啉和其他抑制剂对瘙痒病的预防和治疗活性，并确定了一种对已建立的中枢神经系统感染有效的药物；6)继续将鹿和麋鹿的慢性衰减性疾病适应于仓鼠和表达PrP-sen的转基因小鼠；7)利用氨基酸衍生化和质谱分析PrP-res构象；8)分析PrP突变对PrP向PrP-res转化的影响

Transmissible spongiform encephalopathies (TSEs or prion disease) are fatal neurodegenerative diseases such as scrapie, Creutzfeldt-Jakob disease (CJD), bovine spongiform encephalopathy and chronic wasting disease (CWD). Our project is aimed at understanding and blocking the conversion of normal PrP (PrP-sen) to PrP-res, the abnormal form of prion protein (PrP) that appears to underlie TSE transmission and pathogenesis. We have employed a wide variety of cell biological, biochemical, biophysical and in vivo experimental approaches. Over the last year we have 1)continued the identification and isolation of the most and smallest infectious units of TSE diseases using field flow fractionation, light scattering, electron microscopy and electophoresis, 2) discovered degenerate phosphorothioate oligonucleotides as potent PrP-res inhibitors and anti-TSE drugs in vitro and in vivo, 3)characterized the process by which PrP-res invades neurons and glial cells, initiates infection, and is transported along neuronal processes to extremities where contact is made with other cells, using confocal microscopy, 4) developed the first tissue culture cell line infected with CWD and identified the first inhibitors of CWD replication, 5) continued to test porphyrins and other inhibitors for prophylactic and therapeutic activities against scrapie in animals, and identified one with efficacy against an established central nervous system infection, 6) continued to adapt chronic wasting disease from deer and elk to hamsters and transgenic mice expressing hamster PrP-sen, 7) probed PrP-res conformation using amino acid derivitization and mass spectrometry, 8) analyzed the effects of PrP mutations on PrP conversion to PrP-res