Intrabody Therapy in Drosophila for Huntington's Disease

描述(申请人提供)：我们工作的总体目标是治愈亨廷顿病(HD)。这项建议试图使用已建立的HD果蝇模型来评估针对Huntingtin(HTT)蛋白的新的细胞内抗体(体内)。在组织培养细胞和脑切片制备方面的工作已经证明，体内可以改善与表达突变形式的HTT相关的病理。我们最近已经证明，我们的原型抗HTT体内C4scFv，部分抑制了果蝇功能神经系统中的HD病理。抑制的程度是可以量化的，允许快速评估新的体内，以改善我们现有的体内的疗效。此外，通过果蝇中可用的无偏见的功能遗传筛选，可以阐明体内/靶点相互作用触发的细胞过程，从而导致疾病纠正。识别参与体内依赖的病理纠正的细胞通路将使我们能够设计策略来加强体内治疗以及预测潜在的有害副作用。最后，我们将开发用于测试苍蝇体内的新模型。目前，体内的HTT转基因和致病的HTT转基因从胚胎早期开始共表达。在人类中，体内治疗很可能在疾病发作后在成年人身上开始。我们的新模型将允许在疾病发作后的任何时间测试体内疗效。如果成功，这将提供体内作为疾病治疗方法的验证，并推动在脊椎动物模型中进行临床前试验。

DESCRIPTION (provided by applicant): The overall goal of our work is to cure Huntington's Disease (HD). This proposal seeks to evaluate new intracellular antibodies (intrabodies) directed against the Huntingtin (htt) protein using an established Drosophila model for HD. Work in tissue culture cells and brain slice preparations has demonstrated that intrabodies can ameliorate pathology associated with expression of mutant forms of the htt. We have recently demonstrated that our prototypical anti-htt intrabody, C4 scFv, partially suppresses HD pathology in the functioning nervous system of Drosophila. The degree of suppression is quantifiable, allowing the rapid evaluation of new intrabodies for improved efficacy over our existing intrabody. Additionally, through unbiased functional genetic screens available in Drosophila, the cellular processes triggered by the intrabody/target interaction that result in disease correction can be elucidated. Identification of cellular pathways involved in intrabody-dependent correction of pathology will allow us to devise strategies to augment intrabody therapy as well as predict potential deleterious side effects. Finally, we will develop new models for testing intrabodies in flies. Currently, the intrabody and disease-causing htt transgene are co-expressed from early embryonic stages onward. In humans, therapy with intrabodies would likely commence in adults after the onset of diseases. Our new model will allow testing of intrabody efficacy at any time subsequent to the onset of disease. If successful this will provide validation of intrabodies as disease therapeutics and impetus to proceed to preclinical testing in vertebrate models of HD.