Intracellular tools to study specific misfolded protein variants in human disease

描述（由申请人提供）：蛋白质的细胞内错误折叠或错误加工是许多人类疾病（包括神经退行性疾病、糖尿病和癌症）中的关键问题。特异性靶向和原位操纵单个蛋白质变体的能力将是治疗这些疾病的有力工具。抑制性RNA技术彻底改变了细胞研究，提供了选择性抑制单个基因表达的能力。然而，许多疾病和细胞过程是由异常的蛋白质折叠或加工，或由蛋白质复合物的形成引起的，这些事件不能被RNAi控制。RNAi技术的一个强有力的补充方法是能够选择性地靶向和操纵细胞内错误折叠、错误加工或复合的蛋白质形式。在这项提案中，我们将开发可以使用强大的细胞内蛋白质框架结合特定蛋白质变体的试剂。可以添加不同的标签以靶向特定的细胞位置和途径，包括用于蛋白质降解的蛋白酶体和溶酶体、用于细胞内化的内体靶向和用于蛋白质去除的细胞分泌。可以解决各种非常令人兴奋的应用，包括：1）靶向与包括神经变性疾病、糖尿病和癌症在内的人类健康问题有关的错误折叠和错误加工的蛋白质，2）靶向细胞信号传导途径以控制患病细胞中的凋亡或干细胞中的分化，3）开发可以促进原位正确蛋白质折叠的分子伴侣样试剂，和4）靶向参与复制或装配的病毒蛋白质，以提供控制病毒感染的手段，特别是在脑组织中。该项目的总体目标是开发能够容易地被修饰以在细胞内结合选定的靶蛋白变体并以可预测的方式操纵该靶以实现期望结果的试剂。这种操纵特定细胞内靶点的能力可以对许多健康问题产生深远的影响。 公共卫生相关性：拟议的研究将开发基于重组蛋白的工具，可用于细胞内减轻由靶蛋白的错误折叠或错误加工引起的毒性，控制病变细胞如肿瘤或病毒感染细胞的凋亡，或控制细胞分化和蛋白质折叠。这项研究将有潜在的应用研究，诊断，或治疗疾病引起的异常蛋白质形式，包括阿尔茨海默氏症，帕金森氏症，亨廷顿氏症，糖尿病和癌症。

DESCRIPTION (provided by applicant): Intracellular misfolding or misprocessing of proteins is a critical problem in numerous human diseases including neurodegenerative disorders, diabetes and cancer. The ability to specifically target and manipulate individual protein variants in situ would be a powerful tool in treating these diseases. Inhibitory RNA techniques have revolutionized cell studies, providing the ability to selectively inhibit expression of single genes. Many diseases and cellular processes however result from aberrant protein folding or processing, or from formation of protein complexes, events which cannot be controlled by RNAi. A powerful complementary approach to RNAi techniques would be the ability to selectively target and manipulate a misfolded, misprocessed or complexed protein form intracellularly. In this proposal we will develop reagents that can bind specific protein variants using robust intracellular protein frameworks. Different tags can be added to target specific cell locations and pathways including proteasomes and lysosomes for protein degradation, endosomal targeting for cell internalization, and cell secretion for protein removal. A variety of very exciting applications can be addressed including: 1) targeting misfolded and misprocessed proteins involved in human health problems including neurodegenerative diseases, diabetes and cancer, 2) targeting cell signaling pathways to control apoptosis in diseased cells or differentiation in stem cells, 3) developing chaperone-like reagents that can promote correct protein folding in situ, and 4) targeting viral proteins involved in replication or assembly to provide a means to control viral infection especially in brain tissue. The overall goal of this project is to develop reagents that can be readily modified to intracellularly bind a selected target protein variant and manipulate that target in a predictable manner to achieve a desired result. This ability to manipulate specific intracellular targets can have profound impact on numerous health problems. PUBLIC HEALTH RELEVANCE: The proposed research will develop recombinant protein based tools that can be used to intracellularly alleviate toxicity caused by misfolding or misprocessing of a target protein, to control apoptosis in diseased cells such as tumors or virally infected cells, or to control cell differentiation and protein folding. This research will have potential applications for studying, diagnosing, or treating diseases caused by aberrant protein forms including Alzheimer's, Parkinson's, Huntington's, diabetes, and cancer.