Intrabodies as novel neurological therapeutics

体内作为新型神经治疗方法

• 批准号: 7019240
• 负责人: ANNE MESSER
• 金额: $ 31.46万
• 依托单位: WADSWORTH CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-15 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7019240
• 关键词: Huntington' s disease; Lentivirus; adeno associated virus group; antibody; biotherapeutic agent; drug discovery /isolation; gene delivery system; genetic screening; immunoglobulin structure; intracellular; laboratory mouse; nerve /myelin protein; protein engineering; transfection /expression vector

DESCRIPTION (provided by applicant): The goal of this proposal is to optimize engineered intracellular antibodies (intrabodies) as novel clinical reagents and drug discovery tools for the treatment of Huntington's Disease (HD), with broad, long-term relevance to other neurodegenerative disorders caused by misfolded proteins. Intrabodies use the target specificity of antibodies to form complexes with intracellular proteins, and are already in clinical trials for treatment of cancers and AIDS. The research design starts with in vivo testing with a single-chain Fv anti-huntingtin (htt) intrabody (scFv C4) that has shown significant rescue of HD phenotypes in cell lines, organotypic slice cultures and a Drosophila HD model; plus a newer single domain intrabody (VL 12.3) that shows even stronger anti-htt aggregation properties in situ. Delivery of the intrabody genes will utilize a non-primate lentivirus, Equine Infectious Anemia Virus (EIAV), with either a VSVG or Rabies-g envelope, as one gene therapy vector, with some experiments to compare with delivery using AAV vectors provided by a collaborator. Quantitative assays of abnormal nuclear htt accumulation and aggregation, DARPP- 32 levels, and open field activity behavior will be used to assess the efficacy of the intrabodies delivered to the brains of Exon 1 transgenic (R6/1) and Hdh knock-in (Q111) mouse models on the same inbred genetic background. Simultaneously, screening and testing of a small pool of newer intrabodies will be done using anti-aggregation, protection, and toxicity assays in neuronal cell lines. The most successful of the new intrabodies will then be tested as above. If correction is incomplete with individual intrabodies, combination therapies will be tested in cells and in vivo. At the end of these studies, we will have established the optimal characteristics of intrabodies for eventual HD therapeutics and further drug discovery. These approaches should also be generally applicable for other neurodegenerative diseases that result from abnormal protein folding and accumulation, including Alzheimer's, Parkinson's, and prion diseases.

描述（由申请人提供）：该提案的目的是优化工程的细胞内抗体（内形态）作为治疗亨廷顿氏病（HD）（HD）的新型临床试剂和药物发现工具，与其他与其他神经退行性疾病相关，与其他神经退行性疾病有关。遗产使用抗体的靶特异性与细胞内蛋白质形成复合物，并且已经在临床试验中以治疗癌症和艾滋病。研究设计始于单链FV抗Huntingtin（HTT）内部（SCFV C4）的体内测试，该临床（SCFV C4）显示了细胞系，器官切片培养物和果蝇HD模型中HD表型的大量营救；再加上较新的单域内部机构（VL 12.3），该域在原位显示了更强的抗HTT聚集特性。内部基因的递送将利用非青春期的慢病毒，马传染性贫血病毒（EIAV），带有VSVG或Rabies-g信封作为一种基因治疗载体，并使用协作者提供的AAV矢量进行了一些实验。异常的核HTT积累和聚集，DARPP-32水平以及开放式活动行为的定量测定将用于评估递送给外显子1转基因（R6/1）和HDH敲入（Q111）小鼠模型的外显子1转基因（R6/1）的内体的功效。同时，将使用神经元细胞系中的抗聚集，保护和毒性测定进行筛查和测试。然后，最成功的新体内形态将如上测试。如果校正与单个内遗嘱的校正不完整，则将在细胞和体内测试组合疗法。在这些研究结束时，我们将确定最终HD治疗剂内生内形态的最佳特征和进一步的药物发现。这些方法通常也应适用于其他神经退行性疾病，这些疾病是由异常的蛋白质折叠和积累引起的，包括阿尔茨海默氏症，帕金森氏症和prion疾病。