Biochemistry Of Scrapie Pathogenesis

痒病发病机制的生物化学

• 批准号: 7192913
• 负责人: BYRON CAUGHEY
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7192913
• 关键词: SDS polyacrylamide gel electrophoresis; biochemistry; circular dichroism; confocal scanning microscopy; hamsters; laboratory mouse; light scattering; matrix assisted laser desorption ionization; molecular pathology; neuropathology; prions; protein purification; scrapie; tissue /cell culture

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

可传播的海绵脑病（TSE或PRION疾病）是致命的神经退行性疾病，例如Crapie，Creutzfeldt-Jakob疾病（CJD），牛海绵状脑病疾病和慢性浪费疾病（CWD）。我们的项目旨在理解和阻止正常PRP（PRP-SEN）向PRP-RES的转化，PRP-RES是Prion蛋白（PRP）的异常形式，似乎是TSE传播和发病机理的基础。我们采用了多种细胞生物学，生化，生物物理和体内实验方法。 在过去的一年中，我们有1）继续鉴定和隔离最小和最小的TSE疾病单位，使用场流分馏，光散射，电子显微镜和电触发性，2）发现，发现了磷酸胸酯寡核苷酸的退化磷酸硫硫代寡核苷酸，作为有效的PRP抑制剂和抗抗抑制剂和抗抗抗素的效果，3）vivo，3）和神经胶质细胞，引发感染，并沿着神经元过程运输到与其他细胞接触的肢体，使用共焦显微镜，4）发展了第一个被CWD感染的组织培养细胞系，并确定CWD复制的第一个抑制剂，5）继续测试卟啉和其他抑制剂，并鉴定出对预测性的侵蚀性，并鉴定出对抗脑电的影响，并鉴定出侵蚀性，并鉴定出侵蚀性，并鉴定出对症状的侵蚀性，并确定了反应性的侵蚀性。建立的中枢神经系统感染，6）继续适应从鹿和麋鹿到仓鼠和表达仓鼠PRP-SEN的转基因小鼠的慢性浪费疾病，7）使用氨基酸衍生化和质谱法探测了PRP-RES构象，8）分析了PRP突变对PRP转化的影响PRP-rpp-rp-rp-res-res-res spectration。