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

基本信息

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
项目类别：
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

项目摘要

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.

项目摘要/摘要 E2F转录程序，控制细胞周期的承诺和进展在神经元中DNA损伤后，细胞被上调。神经元DNA损伤和细胞周期失调是神经退行性的特征，也与精神疾病有关。虽然 E2F在增殖过程中的功能已经进行了广泛的研究，E2F在非增殖细胞中的作用，这样与神经元一样，受到关注的关注。新兴证据表明E2F在DNA损伤中起作用维修，独立于其在细胞周期进入中的作用。这种角色在神经元中可能尤其重要，独特的容易受到DNA损伤的影响。尽管E2F诱导与分辨率的DNA损伤有关神经元的散装细胞制剂，这种批量分析无法解释潜在混淆异质性在样品中。单细胞分析对于检查E2F活性与DNA之间的关系是必要的损坏并将E2F动力学与同一细胞中的功能结果联系起来。 Meyer Lab专门研究高通量显微镜数据的单细胞分析，实验室具有为此，最近开发了E2F活性的荧光生物传感器。使用活体显微镜进行定量显微镜 E2F活性和DNA损伤的固定细胞读数将允许映射到细胞命运在数千个细胞中单细胞分辨率的结果。我的假设是，在有丝分裂后神经元中，E2F为可逆地激活以驱动DNA修复，而无需DNA复制后，无dna损伤；但是，如果 DNA损伤激活E2F超出S相进入阈值，E2F诱导DNA复制和凋亡。通过表征如何在神经元中调节E2F程序及其对DNA的贡献损害维修，这项研究将支持鉴定保留遗传的潜在治疗目标以遗传毒性应激为特征的脑疾病的完整性。该研究项目是我作为独立职业培训的重要组成部分研究者采用高通量单细胞方法研究复杂疾病的生物学。我的长期术语目标是成为一名医师科学家，作为精神科医生练习，同时也经营基础科学实验室在学术医院。该提案中概述的计划以及我的赞助商论文的指导委员会以及TRI-I MD-PHD计划的领导，将帮助我实现这些职业愿望。