Intrabodies as novel neurological therapeutics

体内作为新型神经治疗方法

基本信息

批准号：
7674342
负责人：
ANNE MESSER
金额：
$ 7.78万
依托单位：
WADSWORTH CENTER
依托单位国家：
美国
项目类别：
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/7674342
关键词：
Acquired Immunodeficiency Syndrome Affinity Alzheimer&apos s Disease Amino Acids Animals Antibodies Antibody Specificity Behavior Behavioral Binding Biological Assay Brain Brain region Cell Line Cell model Cells Characteristics Class Classification Clinical Clinical Trials[{..}] Cancer Treatment Combined Modality Therapy Complex Corpus striatum structure Cultured Cells DARPP 32 Data Dependovirus Disease Disease model Drosophila genus Engineering Epitopes Equine Infectious Anemia Equine Lentiviruses Exons Functional disorder Gene Delivery Gene Transduction Agent Generations Genes Genetic Goals Huntington Disease Immunoglobulin Fragments In Situ Individual Investigation Knock-in Mouse Length Libraries Long-Term Effects Malignant Neoplasms Mammalian Cell Modeling Molecular Genetics Mus N-terminal Neurodegenerative Disorders Neurologic Neurologic Dysfunctions Neurologic Mutants Mice Neurons Nuclear Parkinson Disease Pathogenesis Primates Prion Diseases Property Proteins Protocols documentation Rabies Reagent Recombinants Research Design Safety Screening procedure Slice Specificity Staging Subfamily lentivirinae Surface System Testing Therapeutic Toxic effect Transgenic Mice Transgenic Organisms Viral Virus Work Yeasts adeno-associated viral vector aged antibody engineering congenic disease phenotype drug discovery experience human Huntingtin protein human disease in vivo innovation mouse model mutant novel novel strategies polyglutamine protein misfolding protein protein interaction research study small molecule tool viral gene delivery

项目摘要

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.

该提案的目的是将工程的细胞内抗体（内形态）优化为新颖用于治疗亨廷顿氏病（HD）的临床试剂和药物发现工具，与错误折叠蛋白引起的其他神经退行性疾病的长期相关性。遗嘱内的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进行癌症和艾滋病的治疗。研究设计始于用单链FV抗狩猎蛋白（HTT）内遗嘱（SCFV C4）进行体内测试，已显示细胞系，器官型切片和果蝇HD中高清表型的大量营救模型;加上一个较新的单域内部遗嘱（VL 12.3），显示出更强的抗HTT 原位聚集属性。递送基因的递送将利用非青春期的慢病毒，马有VSVG或Rabies-G信封，作为一种基因，马传染性贫血病毒（EIAV）治疗载体，进行了一些实验，可以与AAV向量进行比较合作者。异常核HTT积累和聚集的定量测定，darpp- 32个级别，开放式活动行为将用于评估内际的功效传递到外显子1转基因（R6/1）和HDH敲入（Q111）鼠标模型的大脑相同的近交遗传背景。同时筛选和测试一小部分新池内部将使用抗聚集，保护和毒性测定在神经元细胞中进行线。然后，最成功的新体内形态将如上测试。如果校正是与单个内形式不完整，组合疗法将在细胞和体内进行测试。在这些研究的终结，我们将确定内构的最佳特征最终的高清治疗和进一步的药物发现。这些方法也应该通常适用于由异常蛋白质折叠和累积，包括阿尔茨海默氏症，帕金森氏症和病毒疾病。