Molecular impact of endolysosomal dysfunction on neuron-glia communication pathways

内溶酶体功能障碍对神经元-胶质细胞通讯途径的分子影响

• 批准号: 10538113
• 负责人: Nader Francis Morshed
• 金额: $ 6.72万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-29 至 2025-04-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10538113
• 关键词: Address; Affect; Aging; Alzheimer' s Disease; Alzheimer' s disease pathology; Alzheimer' s disease patient; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Astrocytes; Binding; Binding Proteins; Brain; Brain region; Cell physiology; Cell surface; Cells; Communication; Complement; Complement component C4; Complex; Dementia; Disease Progression; Endosomes; Environment; Event; Functional disorder; Genes; Genetic; Genetic Engineering; Genetic Risk; Genetic Transcription; Genetic study; Genetically Engineered Mouse; Gliosis; Goals; Healthcare Systems; Hippocampus (Brain); Human; Impairment; Inflammation; Lead; Learning; Link; Mediating; Microglia; Molecular; Mus; Mutate; Mutation; Nerve Degeneration; Neurobiology; Neurodegenerative Disorders; Neuroglia; Neuroimmune; Neurons; Organelles; Pathogenesis; Pathogenicity; Pathology; Pathway interactions; Patients; Production; Proteins; Proteome; Proteomics; Recycling; Research; Risk; Role; Signal Pathway; Signal Transduction; Synapses; System; Technology; Testing; Time; amyloid precursor protein processing; brain cell; cell type; conditional knockout; cytokine; dementia risk; experimental study; glial activation; human model; human pluripotent stem cell; in vivo; insight; lens; macrophage; molecular pathology; novel; prevent; protein aggregation; protein transport; receptor; receptor binding; risk variant; scaffold; secretion process; stem cell model; tau Proteins; tool; trafficking; trans-Golgi Network

Project Abstract Alzheimer’s disease (AD) is a neurodegenerative disease and the most common form of dementia worldwide. Despite decades of research, there are few therapies that can delay or prevent AD progression. Retrograde trafficking through retromer-dependent cargo recognition has emerged as a critical cellular process that is mutated or disrupted in patients with AD and other forms of dementia. Conditional knockout of retromer genes in neurons leads to increased secretion of Tau and Amyloid β (Aβ), hallmark protein pathologies linked to AD. This milieu of neuronal-secreted factors leads to inflammation in microglia and astrocytes, two glial cell types thought to influence the progression of neurodegeneration. In this proposal, I aim to study the cascade of events linking neuronal retromer disruption to glial inflammation, characterizing the specific cell state changes involved, and identify the key factors that mediate this effect. I will address this aim using genetically engineered stem cell- derived models of human neurons, microglia, and astrocytes. Microglia also express retromer components and upregulate these factors in AD, yet there are few studies of retromer function specifically in microglia. In Aim 2 I will therefore explore the effects of retromer-related mutations specifically on microglia in the context of early aging in mice, a comparable time point to when dementia manifests in patients with these mutations. I will additionally utilize stem cell models to dissect the functional and signaling changes that are induced in microglia with retromer disruption. Finally, although there have been several studies looking at the effects of retromer on specific receptors, little is known about the systems-level effects of retromer mutations on protein trafficking to the endosomes. To identify retromer-dependent signaling pathways that may be pathogenic, I have developed novel proteomics tools to quantify endosomal proteome changes and will use these tools to compare the effects of different retromer mutations on neuronal and microglial endosomes. The ultimate goal is to understand how retromer disruption affects brain cell states and leads to pathogenic signaling changes.

项目摘要 阿尔茨海默病 (AD) 是一种神经退行性疾病，也是全世界最常见的痴呆症。 尽管进行了数十年的研究，但很少有疗法可以延缓或预防 AD 进展。逆行 通过依赖逆转录酶的货物识别进行贩运已成为一个关键的细胞过程， AD 和其他形式的痴呆症患者体内的突变或破坏。条件性敲除retromer基因 神经元中的 Tau 蛋白和 β 淀粉样蛋白 (Aβ) 分泌增加，这是与 AD 相关的标志性蛋白质病理。 这种神经元分泌因子的环境会导致小胶质细胞和星形胶质细胞（两种胶质细胞类型）的炎症 被认为影响神经退行性变的进展。在这个提案中，我的目标是研究一系列事件 将神经元逆转录酶破坏与神经胶质炎症联系起来，描述所涉及的特定细胞状态变化， 并确定介导这种效应的关键因素。我将使用基因工程干细胞来实现这个目标- 人类神经元、小胶质细胞和星形胶质细胞的衍生模型。小胶质细胞还表达逆转录酶成分 在 AD 中上调这些因子，但很少有针对小胶质细胞中逆转录酶功能的研究。目标 2 I 因此，我们将在早期的背景下探讨逆转录酶相关突变对小胶质细胞的影响。 小鼠的衰老，与携带这些突变的患者出现痴呆症的时间点相当。我会 另外利用干细胞模型来剖析小胶质细胞诱导的功能和信号传导变化 与逆转录酶破坏。最后，尽管已经有几项研究探讨了逆转录酶对 特定受体，关于逆转录酶突变对蛋白质运输的系统水平影响知之甚少。 内体。为了识别可能致病的逆转录酶依赖性信号通路，我开发了 新颖的蛋白质组学工具可量化内体蛋白质组变化，并将使用这些工具来比较效果 神经元和小胶质细胞内体上不同逆转录体突变的研究。最终目标是了解如何 逆转录酶破坏会影响脑细胞状态并导致致病信号变化。