Intrabody Therapy of Parkinson's Disease

描述(由申请人提供) 这项提议寻求开发工程细胞内抗体。 (体内)作为潜在的新型临床试剂和药物发现工具 帕金森病(PD)的治疗。内部机构利用 抗体与细胞内蛋白形成复合体的特异性，以及 已经在治疗癌症和艾滋病的临床试验中。最近，我们 发布了这项功能强大的技术的第一个应用程序 神经退行性疾病，表明人类单链抗体在体内， 选自噬菌体展示文库，可中和原位亨廷顿 亨廷顿病(HD)细胞模型中的聚集。通过选择 HD异常折叠膨胀聚谷氨酰胺的邻近表位 蛋白质，我们可以改变异常的蛋白质-蛋白质相互作用 确定突变蛋白的特征。帕金森病患者大脑表现为 丝状细胞内包涵体(路易小体)是它们最引人注目的 病理学。它们似乎是由异常聚集的α-突触核蛋白组成的 这种疾病既有散发性的，也有遗传性的。α-突触核蛋白 到目前为止的蛋白质数据表明，几个候选的靶序列可能 防止突触核蛋白的错误折叠，或用于清除 帕金森氏细胞。我们建议选择抗突触核蛋白单链抗体 体内针对独特的、定义的结构域和形式的a-syn，使用人类 噬菌体展示文库。然后将测试它们的能力 反向报道模型系统中的生理缺陷。型号将包括 几种过度表达野生型α-突触核蛋白以模拟散发的P.D： 在细胞系和脑片培养中的瞬时转基因，继续 转基因果蝇和最终转基因小鼠。所有这些研究都将 善用房屋署研究所得的技术和专业知识 这些研究正在实验室中积极而成功地进行。长期目标包括 通过基因治疗载体给予这些抗体试剂，或作为稳定的， 多功能蛋白质。

DESCRIPTION (provided by applicant) This proposal seeks to develop engineered intracellular antibodies (intrabodies) as novel potential clinical reagents and drug discovery tools for the treatment of Parkinson's disease (PD). Intrabodies make use of the specificity of antibodies to form complexes with intracellular proteins, and are already in clinical trials for treatment of cancers and AIDS. Recently, we published the first application of this powerful technology to a neurodegenerative disease, showing that human single-chain Fv intrabodies, selected from a phage display library, can counteract in situ Huntington aggregation in cellular models of Huntington's Disease (HD). By choosing an epitope adjacent to the abnormally folding expanded polyglutamine of the HD protein, we can alter the abnormal protein-protein interactions that characterize the mutant protein. Parkinson's Disease brains show formation of filamentous intracellular inclusions (Lewy bodies) as their most striking pathology. These appear to be composed of abnormally aggregated alpha-synuclein in both sporadic and hereditary forms of the disease. The alpha-synuclein protein data to date suggest several candidate target sequences that might prevent misfolding of synuclein, or be used to clear abnormal material in Parkinson's cells. We propose to select anti-synuclein single-chain Fv intrabodies against unique, defined domains and forms of a-syn, using human phage display libraries. These will then be tested for their capacity to reverse reported physiological defects in model systems. Models will include several that over-express wild-type alpha-synuclein to mimic sporadic P.D: transient transfections in cell lines and brain slice cultures, moving on to transgenic Drosophila and eventually transgenic mice. All of these studies will take advantage of the technologies and expertise coming out of the HD studies, which are actively and successfully ongoing in the lab. Long-term goals include administering these antibody reagents via gene therapy vectors, or as stable, multi-functional proteins.