Protein homeostasis in a frontotemporal dementia iPSC model

额颞叶痴呆 iPSC 模型中的蛋白质稳态

• 批准号: 10525437
• 负责人: Charleen T Chu
• 金额: $ 43.49万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10525437
• 关键词: ATP phosphohydrolase; Affect; Alzheimer' s Disease; Alzheimer' s disease related dementia; Amyotrophic Lateral Sclerosis; Aphasia; Autophagocytosis; Back; Biological; CRISPR/Cas technology; Cell model; Cells; Chromosomal Stability; Cyclic AMP-Dependent Protein Kinases; Destinations; Disease; Exhibits; Fibroblasts; Foundations; Frontotemporal Dementia; Frontotemporal Lobar Degenerations; Functional disorder; Future; Goals; Golgi Apparatus; Grant; Health; Homeostasis; Human; Inclusion Body Myopathy with Early-Onset Paget Disease; Infrastructure; Lead; Link; Mediating; Modeling; Morphology; Motor Neurons; Musculoskeletal Diseases; Mutation; Myoblasts; N Domain; NF1 gene; Nerve Degeneration; Neurites; Neurodegenerative Disorders; Neurons; PINK1 gene; Parkinson Disease; Parkinsonian Disorders; Pathogenicity; Patients; Phenotype; Phosphotransferases; Play; Primary Progressive Aphasia; Process; Protein Biosynthesis; Protein Secretion; Proteins; Reagent; Role; Source; Stress; Synapses; System; Testing; Transfection; Translating; brain cell; cell type; cofactor; design; disease-causing mutation; endoplasmic reticulum stress; induced pluripotent stem cell; link protein; motor disorder; multisystem proteinopathy; novel therapeutic intervention; pluripotency; protein biomarkers; protein function; protein transport; proteostasis; valosin containing protein mutation; valosin-containing protein

Maintaining protein homeostasis is a particular challenge for neurons, in which a high demand for protein synthesis, folding and transport represents a constant source of stress. Evidence of protein mishandling is observed in nearly all neurodegenerative diseases including the AD-related dementias (ADRD). The AAA- ATPase valosin-containing protein (VCP) plays a central role in maintaining multiple aspects of protein homeostasis. Mutations in VCP have been linked to several forms of frontotemporal lobar degeneration with or without concurrent motor dysfunction and musculoskeletal disease. Yet there is limited understanding of how these mutations affect different aspects of VCP function in neurons. The VCP-T262A mutation causes familial frontotemporal dementia with aphasia and parkinsonism. Preliminary studies in primary neurons transfected with VCP-T262A reveal deficits in dendritic arborization, and patient fibroblasts bearing the endogenous mutation exhibit disrupted secretory function. Reagents to create human neurons and other relevant cell types bearing the endogenous VCP-T262A mutation are critically needed to study pathophysiological mechanisms of disease. In this exploratory project, we will create isogenic pairs of iPSC lines expressing T262A vs. wild type VCP. We will differentiate to cortical neurons to study the effects of this mutation upon ER stress, autophagy and Golgi markers compared to iPSC-derived cells with a mutation in a different VCP functional domain. Our long-range goals are to understand how alterations in VCP contribute to synaptic loss so that this mechanistic understanding can be translated into new therapeutic approaches.

维持蛋白质稳态对于神经元来说是一个特殊的挑战，因为神经元对蛋白质的需求很高 合成、折叠和运输是持续的压力源。蛋白质处理不当的证据是 在几乎所有神经退行性疾病中都观察到这一现象，包括 AD 相关痴呆 (ADRD)。 AAA- 含 ATP 酶缬洛辛蛋白 (VCP) 在维持蛋白质的多个方面发挥着核心作用 体内平衡。 VCP 突变与多种形式的额颞叶变性有关 或没有并发运动功能障碍和肌肉骨骼疾病。但人们的了解还有限 这些突变如何影响神经元中 VCP 功能的不同方面。 VCP-T262A突变导致 家族性额颞叶痴呆伴失语症和帕金森症。原代神经元的初步研究 转染 VCP-T262A 的患者显示树突状树枝化缺陷，并且患者成纤维细胞具有 内源性突变表现出分泌功能破坏。创建人类神经元和其他物质的试剂 迫切需要研究携带内源性 VCP-T262A 突变的相关细胞类型 疾病的病理生理机制。在这个探索性项目中，我们将创建 iPSC 的同基因对 表达 T262A 的细胞系与表达野生型 VCP 的细胞系。我们将分化为皮质神经元来研究其影响 与具有以下突变的 iPSC 衍生细胞相比，ER 应激、自噬和高尔基体标记物发生突变 不同的VCP功能域。我们的长期目标是了解 VCP 的变化如何 导致突触损失，因此这种机制理解可以转化为新的治疗方法 接近。