The Neurodevelopmental Consequences of Genomic Stress

基因组压力的神经发育后果

• 批准号: 9911484
• 负责人: Nadine Michel
• 金额: $ 3.35万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-30 至 2021-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9911484
• 关键词: Adult; Affect; Aging; Alzheimer' s Disease; Animal Model; Aphidicolin; Apoptosis; Astrocytes; Autopsy; Biological Assay; Biological Models; Brain; Camptothecin; Cell Cycle; Cell Cycle Arrest; Cell Cycle Checkpoint; Cells; Copy Number Polymorphism; DNA Damage; DNA Double Strand Break; DNA Repair; Data; Development; Disease; Dose; Exhibits; Exposure to; Fibroblasts; Flow Cytometry; Frequencies; Genes; Genetic Transcription; Genomics; Goals; Hour; Human; Image; Immunoblotting; Individual; Inflammatory; Knowledge; Laboratories; Lead; Life; Long-Term Effects; Longevity; Longitudinal Studies; Measures; Mediating; Metabolic; Mutagens; Nerve Degeneration; Neurodegenerative Disorders; Neuronal Differentiation; Neurons; Nucleotides; Parkinson Disease; Pathologic; Patients; Phenotype; Phosphorylation; Play; Population; Process; Production; Proteins; Pseudogenes; Resistance; Role; Running; Signal Pathway; Signal Transduction; Somatic Cell; Somatic Mutation; Stress; Study models; TP53 gene; Testing; Tissues; Transcript; Transgenic Mice; Type I DNA Topoisomerases; Variant; Western Blotting; aged; aging brain; base; cell type; cytokine; early onset; genomic platform; human model; in vitro Model; induced pluripotent stem cell; knock-down; mouse model; nerve stem cell; neurodegenerative phenotype; neurodevelopment; neurogenesis; neuroinflammation; neuron development; neuroregulation; normal aging; prevent; programs; relating to nervous system; replication stress; resistance mechanism; response; single cell analysis; single-cell RNA sequencing; small hairpin RNA; stem cells; stressor; tool

ABSTRACT: The abnormal accumulation of DNA damage in neurons is a shared pathological feature among neurodegenerative diseases (Alzheimer's disease, Parkinson's disease) and it is also correlated with normal aging. Transgenic mouse models further demonstrate a specific requirement for DNA repair during neurogenesis. It is also known that genomic stress, arising from ongoing transcription and metabolic byproducts, plays a role in aging because it contributes to the accumulation of DNA damage. Previously, studying aging in the human brain has been incredibly challenging; post-mortem tissue is difficult to acquire and is not amenable to longitudinal studies. Furthermore, studies of aging have typically looked at model organisms later in life, meaning that few have assessed how stress exposure impacts the development of the brain and how that ultimately affects the aging process. With the development of human-induced pluripotent stem cells (hiPSCs), we now have an in vitro model of human neurodevelopment that provides a tool to study models of normal and pathological neurodevelopment. The goal of this study is to determine how genomic stress and subsequent DNA damage alters the diversity among hiPSC-derived neural progenitor cells (NPCs) and their progeny. Aim one seeks to determine how NPCs are resistant to replicative stress and whether this resistance is unique to NPCs compared to isogenic fibroblasts, astrocytes, and neurons. Multispectral imaging flow cytometry, immunoblotting, and cell cycle analysis will all be used to elucidate the mechanisms of resistance by investigating DNA damage repair proteins. This aim is founded on preliminary data indicating that NPCs are resistant to replication stress and particularly susceptible to transcription associated genomic stress; showing an increase in DNA damage, cell cycle arrest, and DNA damage signaling activation. Aim two seeks to determine if genomic stress promotes neuroinflammatory phenotypes through single cell RNA-seq and the human Luminex assay. hiPSC-based neurogenesis provides a human model system to understand genomic stress related mechanisms in aging and neurodegeneration. This study will define how neurodevelopmental stress may predispose subsets of neurons for later neurodegeneration through neuroinflammatory phenotypes, and ultimately lead to consequent studies focused on protecting NPCs and their progeny from genomic stress to prevent accelerated aging and neurodegenerative disease.

抽象的： 神经元中DNA损伤的异常积累是一种共同的病理特征 神经退行性疾病（阿尔茨海默氏病，帕金森氏病），也与正常 老化。转基因小鼠模型进一步证明了对DNA修复的特定要求 神经发生。还知道，持续转录和代谢副产品引起的基因组应激， 由于它有助于DNA损伤的积累，因此在衰老中起作用。以前，研究衰老 人的大脑一直具有挑战性。验尸组织很难获取，并且不适合 进行纵向研究。此外，对衰老的研究通常会在以后的生活中研究模型生物， 这意味着很少有人评估压力暴露如何影响大脑的发展以及如何影响 最终会影响衰老过程。 随着人类诱导的多能干细胞（HIPSC）的发展，我们现在有一个体外模型 人类神经发育，提供了一种研究正常和病理神经发育模型的工具。 这项研究的目的是确定基因组应力和随后的DNA损伤如何改变多样性 在HIPSC衍生的神经祖细胞（NPC）及其后代中。 AIM一个人试图确定NPC如何 对复制应激具有抗性，并且与等源成纤维细胞相比，这种抗性是否是NPC所独有的， 星形胶质细胞和神经元。多光谱成像流式细胞术，免疫印迹和细胞周期分析将全部 用于通过研究DNA损伤修复蛋白来阐明电阻机制。这个目标是 建立在初步数据上，表明NPC对复制应力有抵抗力，尤其易感性 转录相关的基因组应激；显示DNA损伤，细胞周期停滞和DNA的增加 损坏信号激活。目标两个试图确定基因组应力是否促进神经炎症 通过单细胞RNA-seq和Human Luminex分析的表型。基于HIPSC的神经发生提供了 人类模型系统了解衰老和神经变性中的基因组应力相关机制。这 研究将定义神经发育应激如何使神经元的神经元子集偏爱后来的神经变性 通过神经炎症表型，最终导致研究专注于保护NPC 以及它们来自基因组应激的后代，以防止加速衰老和神经退行性疾病。