Chaperone Dysfunction in Myopathy: Connecting Yeast Genetics with Mouse Models

肌病中的伴侣功能障碍：将酵母遗传学与小鼠模型联系起来

• 批准号: 10434651
• 负责人: HEATHER L TRUE-KROB
• 金额: $ 66.7万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10434651
• 关键词: Affect; Amyloid; Animal Model; Biological Models; Cell model; Cells; Client; Data; Defect; Degenerative Disorder; Desmin; Disease; Functional disorder; Gene-Modified; Goals; Grant; Guanine Nucleotide Exchange Factors; Heat shock proteins; Heat-Shock Proteins 70; Human; In Vitro; Inherited; Intermediate Filaments; Knock-in Mouse; Lead; Limb-Girdle Muscular Dystrophies; Mammalian Cell; Molecular; Molecular Chaperones; Molecular Conformation; Mus; Muscle; Muscle Cells; Muscle Fibers; Muscular Dystrophies; Mutation; Myofibrils; Myopathy; Nucleotides; Pathogenesis; Pathogenicity; Pathology; Patients; Phenotype; Prions; Process; Proteins; RNA-Binding Proteins; Reporting; Role; Site; Skeletal Muscle; System; Testing; Therapeutic; Therapeutic Intervention; Toxic effect; Transgenic Mice; Work; Yeasts; base; conformer; disease phenotype; disease-causing mutation; gene correction; heat-shock proteins 40; in vivo; induced pluripotent stem cell; inhibitor; model development; mouse model; mutant; novel therapeutic intervention; novel therapeutics; prion-like; protein TDP-43; protein aggregation; proteostasis; small molecule; success; sup35; therapeutic target; tool; yeast genetics

Heat shock proteins (HSPs) or protein chaperones are essential to maintain cellular protein homeostasis. Mutations of proteins within this network can lead to degenerative diseases including muscular dystrophy. We have now identified mutations in two domains of an HSP40 co-chaperone, DNAJB6, in an autosomal dominantly inherited Limb Girdle Muscular Dystrophy (LGMDD1). Our central hypothesis is that LGMDD1 mutations alter the conformation of DNAJB6 such that the interface between these two domains is disrupted, leading to myofibril disorganization and muscle dysfunction. Based on our results, we further hypothesize that all LGMDD1 mutations in DNAJB6 can be rescued by perturbation of Hsp70 interaction or activity. Thus, our approach is to employ all of the tools we have developed to test these hypotheses. In order to tackle this challenging problem, we will explore DNAJB6 chaperone function and dysfunction utilizing multiple systems and the expertise of two PIs. Aim 1 will explore whether manipulating DNAJB6:HSP70 interactions rescues LGMDD1 phenotypes in mouse models and human cells. Aim 2 will determine the effect of newly identified LGMDD1-associated DNAJB6 J- domain mutations on chaperone activity in cellular models. Aim 3 will determine the effect of LGMDD1 mutations on chaperone and co-chaperone function in vivo and in vitro. Our preliminary data suggest that we have identified additional therapeutic targets that may be better tolerated in patients than global inhibitors of Hsp70. The overarching goal of this work is to leverage our transdisciplinary success to develop therapeutic intervention for patients with LGMDD1.

热休克蛋白（HSP）或蛋白伴侣是维持细胞蛋白质 体内平衡这个网络中的蛋白质突变会导致退行性疾病 包括肌肉萎缩症我们现在已经在HSP 40的两个结构域中发现了突变， 常染色体显性遗传肢带型肌营养不良症中的辅助分子伴侣DNAJB 6 （LGMDD1）。我们的中心假设是LGMDD 1突变改变了 DNAJB 6，使得这两个结构域之间的界面被破坏，导致肌原纤维形成。 组织紊乱和肌肉功能障碍。根据我们的研究结果，我们进一步假设，所有 DNAJB 6中的LGMDD 1突变可以通过干扰Hsp 70相互作用或活性来挽救。 因此，我们的方法是使用我们开发的所有工具来测试这些假设。在 为了解决这个具有挑战性的问题，我们将探索DNAJB 6分子伴侣的功能， 利用多个系统和两个PI的专业知识的功能障碍。目标1将探讨 操纵DNAJB 6：HSP 70相互作用在小鼠模型中拯救LGMDD 1表型， 人类细胞。目的2将确定新鉴定的LGMDD 1相关DNAJB 6 J- 结构域突变对细胞模型中伴侣蛋白活性的影响。目标3将决定 LGMDD 1突变对分子伴侣和共分子伴侣在体内和体外的功能。我们 初步数据表明，我们已经确定了其他治疗靶点， 在患者中的耐受性优于Hsp 70的总体抑制剂。这项工作的首要目标是 利用我们跨学科的成功，为患有 LGMDD1.