Identification of factors essential for age-related neuronal health: insights into common mechanisms of neurodegeneration

识别与年龄相关的神经元健康所必需的因素：深入了解神经退行性变的常见机制

• 批准号: 10057170
• 负责人: Brian A Link
• 金额: $ 42.9万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10057170
• 关键词: Address; Adult; Affect; Age; Age Factors; Aging; Alzheimer' s Disease; Ataxia; Autophagocytosis; Axon; Axotomy; Basic Science; Behavioral; Biology; Brain; Cell Death; Cell physiology; Cellular Stress; Code; Demographic Aging; Disease; Engineering; Gene Expression Regulation; Genes; Genetic; Genetic Screening; Genomics; Health; Health Care Costs; Homeostasis; Homologous Gene; Human; Huntington Disease; Image; Individual; Insertional Mutagenesis; Mammals; Measures; Molecular; Monitor; Motor Neurons; Mutation; Natural regeneration; Nerve Degeneration; Nervous system structure; Neuraxis; Neurodegenerative Disorders; Neuroglia; Neuronal Injury; Neurons; Optic Nerve; Optic Nerve Transections; Oxidation-Reduction; Oxidative Stress; Parkinson Disease; Pathway interactions; Phenotype; Population; Predisposition; Process; Proteins; Regulation; Reporter; Research; Retinal Degeneration; Retinal Ganglion Cells; Rodent; Rodent Model; Role; Signal Transduction; Spinal Cord transection injury; Spinal cord injury; Spinocerebellar Ataxias; Stress; Targeted Research; Testing; Therapeutic Intervention; Transgenic Organisms; Zebrafish; age related; base; biological adaptation to stress; cell type; dopaminergic neuron; effective intervention; endoplasmic reticulum stress; experimental study; ganglion cell; genetic strain; genome editing; insight; interest; loss of function mutation; mutant; mutation screening; neuron loss; neuronal survival; neuroprotection; novel; null mutation; prevent; proteostasis; reinnervation; response; social; stress granule; targeted treatment; trafficking; transcriptome sequencing

In a genetic screen conducted in adult zebrafish to select mutations that disrupt age-related neuronal homeostasis, 36 genes were identified. Many of these loci have previously been shown as associated or causative to human neurodegenerations such as Alzheimer's, Huntington's, Parkinson's and Spinocerebellar Ataxia diseases. This proposal describes experiments to address cellular functions of a subset of the genes identified from this screen with unknown mechanisms of action. First, complete loss of function mutations will be generated in this subset though engineered large genomic deletions within coding sequence. Next, these will be analyzed for their impact on neuronal homeostasis by monitoring several cellular stress responsive pathways known to be activated during neurodegeneration including the ER stress/Unfolded Protein Response, Oxidative Stress Response, and Autophagy/Mitophagy. In anticipation of these studies, our group has established imaging-based reporters for each of these processes. In addition, mutants will also be evaluated in the context of several sensitized genetic strains in which these stress-responsive pathways have been compromised. Finally, the autonomy of mutant impact will be investigated through cell-type specific gene disruption in multiple neuronal and non-neuronal cell types. These studies will provide important base-line information and rationale for carrying out next-step analysis in mammalian, including human, homologues that may represent critical targets for therapeutic intervention to promote neuronal health and prevent neurodegeneration.

在一项对成年斑马鱼进行的基因筛选中， 稳态，鉴定了36个基因。这些基因座中的许多先前已显示为相关的或 导致人类神经退化，如阿尔茨海默氏症、亨廷顿氏症、帕金森氏症和脊髓小脑症 共济失调疾病。该提案描述了解决基因子集的细胞功能的实验 从这个屏幕上识别出未知的作用机制。首先，完全丧失功能的突变将 通过编码序列内的工程化大基因组缺失在该亚组中产生。接下来，这些 将通过监测几种细胞应激反应来分析它们对神经元稳态的影响。 已知在神经变性过程中激活的途径，包括ER应激/未折叠蛋白 反应、氧化应激反应和自噬/线粒体自噬。在这些研究的预期中，我们的团队 已经为这些过程中的每一个建立了基于成像的报告器。此外，突变体还将在 在几种致敏遗传菌株的背景下进行了评估，其中这些应激反应途径具有 已经暴露了最后，将通过细胞类型特异性基因来研究突变影响的自主性 破坏多种神经元和非神经元细胞类型。这些研究将提供重要的基线 在哺乳动物包括人类中进行下一步分析的信息和理由， 可能代表治疗干预的关键靶点，以促进神经元健康和预防 神经变性