Understanding the mechanistic role of genome stability pathways in regulating cell homeostasis

了解基因组稳定性途径在调节细胞稳态中的机制作用

基本信息

批准号：
10574614
负责人：
Tony Tung Huang
金额：
$ 42.38万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-15 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10574614
关键词：
Address Area Biological Models Cell Cycle Checkpoint Cell Nucleus Cell Surface Receptors Cells Cytoplasm DNA DNA Damage DNA Repair DNA Repair Disorder DNA biosynthesis DNA replication fork Detection Environment Gene Expression Genome Genome Stability Genomic Instability Genomics Genotoxic Stress Goals Growth Growth Factor Homeostasis Interphase Cell Knowledge Ligation Link Maintenance Mediating Metabolic Microscopy Molecular Nuclear Okazaki fragments Organism PIK3CG gene Pathway interactions Process Proteomics Proto-Oncogene Proteins c-akt Receptor Protein-Tyrosine Kinases Recovery Research Role Signal Pathway Signal Transduction Site Source Stress Techniques Testing Viral cancer therapy cell type extracellular genome integrity genome-wide genome-wide analysis genotoxicity human disease innovation malignant neurologic neoplasms nervous system disorder novel preservation programs response single molecule superresolution microscopy

项目摘要

PROJECT SUMMARY/ABSTRACT To preserve genomic integrity and maintain homeostasis, cells in our body must effectively respond to both exogenous and endogenous sources of DNA damage. How cellular DNA damage contribute to human disease, including neurological disorders and cancer, is a fundamental area of research. For the past thirteen years, my lab has been focused on studies related to the mechanistic basis of genomic instability. The goal of this MIRA application is to address critical gaps in our understanding of genome stability pathways and how they are differentially utilized in dividing versus non-dividing cells for the proper maintenance of cellular homeostasis. By relaying external information from the cell periphery to the nucleus, cell surface receptor tyrosine kinases (RTKs) respond to growth factors via PI3-kinase (PI3K)-AKT signaling to regulate gene expression and thereby promote growth and/or survival. Similarly, DNA damage threatens genome integrity and upon detection within the nucleus elicits DNA damage response (DDR) signaling to aid in DNA repair and cell cycle checkpoints. For our research program, we will address unique mechanisms related to how key DDR factors contribute to extracellular growth factor signaling crosstalk in dividing and non-dividing cells. To establish these mechanisms, we will utilize an array of innovative experimental approaches including genome-wide sequencing, proximity ligation proteomics, super-resolution microscopy and single-molecule tracking in live cells. We will test the hypothesis that the DDR should be viewed as a broader, stress-responsive network linking nuclear and cytoplasmic effectors to maintain physiological homeostasis through intersecting with growth factor signaling pathways. How this is achieved mechanistically will be a major focus of this application. The second project involves addressing how genotoxic stress in dividing cells impacts DNA replication dynamics and to elucidate novel molecular players that regulate replication fork recovery. Based on our innovative technique called Okazaki fragment sequencing (OK-seq), we are able to directly quantify the efficiency of replication fork initiation and termination at specific sites throughout the genome. Using this technique, we will expand our analysis to understand how genotoxic insults and DNA repair deficiencies contribute to site-specific replication fork-mediated DNA breaks using genome-wide analysis. Deciphering the mechanisms that contribute to replication-associated genomic instability may provide new avenues for targeted cancer treatment.

项目概要/摘要为了保持基因组完整性并维持体内平衡，我们体内的细胞必须有效地对两者做出反应 DNA 损伤的外源性和内源性来源。细胞 DNA 损伤如何导致人类疾病，包括神经系统疾病和癌症，是一个基础研究领域。在过去的十三年里，我的实验室一直专注于与基因组不稳定性的机制基础相关的研究。 MIRA 的目标应用程序旨在解决我们对基因组稳定性途径及其运作方式的理解中的关键差距在分裂细胞和非分裂细胞中的利用有差异，以正确维持细胞稳态。经过将外部信息从细胞外周传递到细胞核、细胞表面受体酪氨酸激酶 (RTK) 通过 PI3 激酶 (PI3K)-AKT 信号传导响应生长因子，调节基因表达，从而促进生长和/或生存。同样，DNA 损伤威胁着基因组的完整性，并且在细胞核内检测到时引发 DNA 损伤反应 (DDR) 信号，以帮助 DNA 修复和细胞周期检查点。对于我们的研究计划中，我们将解决与关键 DDR 因素如何促进细胞外生长相关的独特机制分裂和非分裂细胞中的因子信号串扰。为了建立这些机制，我们将利用一系列创新的实验方法，包括全基因组测序、邻近连接蛋白质组学、超分辨率显微镜和活细胞中的单分子追踪。我们将检验 DDR 的假设应被视为一个更广泛的压力响应网络，连接核和细胞质效应器以维持通过与生长因子信号通路交叉实现生理稳态。这是如何实现的从机械上讲，这将是该应用程序的主要焦点。第二个项目涉及解决遗传毒性问题分裂细胞中的压力会影响 DNA 复制动态，并阐明调节的新分子参与者复制叉恢复。基于我们称为冈崎片段测序 (OK-seq) 的创新技术，我们能够直接量化整个过程中特定位点的复制叉起始和终止的效率基因组。使用这种技术，我们将扩展我们的分析，以了解基因毒性损伤和 DNA 是如何使用全基因组分析，修复缺陷会导致位点特异性复制叉介导的 DNA 断裂。破译导致复制相关基因组不稳定性的机制可能提供新的思路癌症靶向治疗的途径。