Noncanonical E2F Regulation in the Neuronal DNA Damage Response

神经元 DNA 损伤反应中的非典型 E2F 调节

• 批准号: 10752078
• 负责人: David Leon Rosenthal
• 金额: $ 5.26万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-15 至 2026-05-14
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10752078
• 关键词: Anxiety Disorders; Apoptosis; Apoptotic; Attention; Basic Science; Biology; Biosensor; Bipolar Disorder; Brain Diseases; CDK2 gene; Career Choice; Cell Cycle; Cell Cycle Progression; Cell Cycle Proteins; Cell Cycle Regulation; Cell Proliferation; Cells; Cellular Assay; Cellular Structures; Complex; DNA Damage; DNA Repair; DNA Replication Induction; DNA biosynthesis; Data; Data Analyses; Disease; Doctor of Philosophy; Etiology; Exposure to; Genetic; Genetic Transcription; Genotoxic Stress; Goals; Heterogeneity; Hospitals; Immunofluorescence Immunologic; Individual; Interphase Cell; Knowledge; Leadership; Link; Longevity; Maps; Measures; Mental disorders; Mentorship; Metabolism; Methods; Microscopy; Mood Disorders; Nerve Degeneration; Neurodegenerative Disorders; Neuronal Dysfunction; Neurons; Outcome; Oxidative Stress; Pathway interactions; Phosphorylation; Physicians; Physiological; Proliferating; Proteins; Psychiatrist; Quantitative Microscopy; Recording of previous events; Regulation; Reporter; Research; Research Personnel; Research Project Grants; Resolution; Role; Running; S phase; Sampling; Schizophrenia; Scientist; Severities; Signal Pathway; Signal Transduction; Stress; Systems Biology; Techniques; Testing; Tissues; Training; Work; career; cell fixing; cell preparation; experience; experimental study; functional outcomes; genome integrity; high throughput analysis; human disease; neuropsychiatric disorder; novel; pharmacologic; postmitotic; preservation; programs; protein expression; repaired; response; single cell analysis; upstream kinase

PROJECT SUMMARY/ABSTRACT The E2F transcriptional program, which controls cell cycle commitment and progression in proliferating cells, is upregulated following DNA damage in neurons. Neuronal DNA damage and cell cycle dysregulation are features of neurodegeneration that have also been associated with psychiatric diseases. Although the function of E2F during proliferation has been extensively studied, the role of E2F in nonproliferating cells, such as in neurons, has received less attention. Emerging evidence indicates that E2F plays a role in DNA damage repair, independent of its role in cell cycle entry. This role may be especially important in neurons, which are uniquely vulnerable to DNA damage. Although E2F induction has been linked to resolution of DNA damage in bulk-cell preparations of neurons, such bulk analyses fail to account for potentially confounding heterogeneity within samples. Single cell analysis is necessary to examine the relationship between E2F activity and DNA damage and to link E2F dynamics to functional outcomes in the same cells. The Meyer Lab specializes in single-cell analysis of high-throughput microscopy data, and the lab has recently developed a fluorescent biosensor of E2F activity for this purpose. Quantitative microscopy using live- and fixed-cell readouts of E2F activity and DNA damage will allow signaling history to be mapped to cell fate outcomes at single-cell resolution in thousands of cells. My hypothesis is that in postmitotic neurons, E2F is reversibly activated to drive DNA repair without DNA replication following sublethal DNA damage; however, if DNA damage activates E2F beyond the threshold for S-phase entry, E2F induces DNA replication and apoptosis. By characterizing how the E2F program is regulated in neurons and how it contributes to DNA damage repair, this study will support the identification of potential treatment targets for preserving genetic integrity in brain disorders characterized by genotoxic stress. This research project represents an important component of my training for a career as an independent investigator, employing high-throughput single-cell methods to study the biology of complex diseases. My long- term goal is to become a physician-scientist, practicing as a psychiatrist while also running a basic science lab in an academic hospital. The plan outlined in this proposal along with the mentorship of my sponsor, thesis committee, and the leadership of the Tri-I MD- PhD program, will help me achieve these career aspirations.

项目总结/摘要 E2 F转录程序，控制细胞周期的承诺和进展，在增殖 在神经元中DNA损伤后上调。神经元DNA损伤与细胞周期失调 是神经变性的特征，也与精神疾病有关。虽然 E2 F在增殖过程中的功能已被广泛研究，E2 F在非增殖细胞中的作用，如 就像神经元一样，受到的关注较少。新的证据表明E2 F在DNA损伤中起作用 修复，独立于其在细胞周期进入中的作用。这种作用在神经元中可能特别重要， 特别容易受到DNA损伤虽然E2 F诱导与DNA损伤的解决有关， 大量的神经元细胞制备，这种批量分析不能解释潜在的混杂异质性 在样品中。单细胞分析是必要的，以检查E2 F活性和DNA之间的关系 并将E2 F动力学与相同细胞中的功能结果联系起来。 迈耶实验室专门从事高通量显微镜数据的单细胞分析，该实验室已 最近开发了E2 F活性的荧光生物传感器用于此目的。定量显微镜使用活的- 固定细胞的E2 F活性和DNA损伤的读数将允许信号历史映射到细胞命运 在数千个细胞中实现单细胞分辨率。我的假设是，在有丝分裂后的神经元中，E2 F是 在亚致死性DNA损伤后，可逆地激活以驱动DNA修复而不进行DNA复制;然而，如果 DNA损伤激活E2 F超过S期进入的阈值，E2 F诱导DNA复制， 凋亡通过描述E2 F程序在神经元中的调节方式以及它如何对DNA产生影响， 损伤修复，这项研究将支持确定潜在的治疗靶点，以保持遗传 以遗传毒性应激为特征的脑部疾病的完整性。 这个研究项目是我作为一个独立的职业培训的重要组成部分 研究人员，采用高通量单细胞方法研究复杂疾病的生物学。我的长- 我的长期目标是成为一名医生兼科学家，在经营一个基础科学实验室的同时，也作为一名精神病医生执业 在一家学术医院。计划概述在这一建议沿着与导师我的赞助商，论文 委员会，和三-I MD-博士课程的领导，将帮助我实现这些职业抱负。