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Shaping DNA Damage Response Networks Via Histone H2A Variants

通过组蛋白 H2A 变体塑造 DNA 损伤反应网络

基本信息

批准号：
9254533
负责人：
Kyle M Miller
金额：
$ 38.53万
依托单位：
UNIVERSITY OF TEXAS AT AUSTIN
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-05 至 2021-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9254533
关键词：
Address BRCA1 gene Biochemical CRISPR/Cas technology Cancer cell line Cell Line Cell Survival Cells Chromatin Chromatin Structure DNA Damage DNA Double Strand Break DNA Repair DNA analysis Data Disease Double Strand Break Repair Environment Epigenetic Process Etiology Gene Expression Genetic Genome Genome Stability Goals Histone H2A Histones Homeostasis Human Human Engineering Individual Knock-out Knowledge Location Maintenance Malignant Neoplasms Mediating Mediator of activation protein Modeling Molecular Pathway interactions Play Production Proteins Proteomics Public Health Radiation Radiation therapy Recruitment Activity Role Shapes Signal Transduction Source Structure System Technology Testing Transcriptional Regulation Translating Variant Work base cancer therapy chemotherapeutic agent drug discovery epigenome epigenomics genetic information genome editing genome integrity homologous recombination improved inhibitor/antagonist innovation insight knock-down loss of function novel novel strategies protein E public health relevance radiation response repaired response small hairpin RNA treatment response tumor progression whole genome

项目摘要

DESCRIPTION (provided by applicant): Chromatin-based DNA damage response (DDR) pathways protect cells from genome and epigenome instability, which are hallmarks of cancer and are thought to drive cancer progression. This proposal addresses questions of fundamental importance for epigenetic and genetic mechanisms involved in cancer. Histone H2A variants are critical components of chromatin but how they promote epigenome and genome stability are poorly understood. For this proposal, we aim to use genetic systems using our histone H2A variant knockout/knockdown engineered human cells with complimentary proteomic analyses of histone variants to answer fundamental questions for how histone H2A variants promote the DDR to maintain genome stability. We also aim to provide insights into how histone H2A variants regulate radiation responses, a therapy often used in cancer treatments. These studies provide innovative and new approaches to these questions that can provide transformative insights into chromatin-based DNA damage responses mediated by histone H2A variants. We will first analyze the DDR function of histone H2A variants in response to radiation, by using human cells deleted or depleted for histone H2A variants including H2AX, H2AZ, macroH2A and H2A.Bbd. Our preliminary results indicate that histone H2A variants play vital and unique roles in these pathways. These studies are poised to provide critical insights into how histone H2A variants regulate the DDR and response to IR. We will then exploit our preliminary analyses identifying candidate protein-interacting factors for each histone H2A variant to identify DDR effector proteins for individual histone H2A variants. Finally, we will test our working hypothesis for how macroH2A promotes DNA repair by HR through the interactions with a new DDR factor that we have identified. These studies will provide an unparalleled view of how histone H2A variants shape the DDR to protect the genome integrity across structurally and functionally diverse chromatin landscapes in human cells. DNA damaging agents are a major class of chemotherapeutic agents including radiotherapy and the epigenome represents exciting new leads for drug discovery. Thus, this work exploits a combination of genetic, biochemical and cellular approaches in human cells to identify key DDR pathways that promote genome and epigenome integrity, as well as responses to cancer-relevant therapies including radiation and PARP inhibitors that can translate to improved treatments for human cancers.

描述（由申请人提供）：基于染色质的DNA损伤反应（DDR）途径保护细胞免受基因组和表观基因组不稳定性的影响，这些不稳定性是癌症的标志，并被认为会推动癌症进展。该提案解决了癌症中涉及的表观遗传和遗传机制的根本重要性问题。组蛋白H2 A变体是染色质的重要组成部分，但它们如何促进表观基因组和基因组稳定性却知之甚少。对于这个提议，我们的目标是使用我们的组蛋白H2 A变体敲除/敲低工程化人类细胞的遗传系统，并对组蛋白变体进行补充蛋白质组学分析，以回答组蛋白H2 A变体如何促进DDR以维持基因组稳定性的基本问题。我们还旨在深入了解组蛋白H2 A变体如何调节辐射反应，这是一种常用于癌症治疗的疗法。这些研究为这些问题提供了创新和新的方法，可以为组蛋白H2 A变体介导的基于染色质的DNA损伤反应提供变革性的见解。我们将首先通过使用组蛋白H2 A变体（包括H2 AX、H2 AZ、macroH 2A和H2A.Bbd）缺失或耗尽的人细胞来分析组蛋白H2 A变体响应于辐射的DDR功能。我们的初步结果表明，组蛋白H2 A变体在这些途径中发挥着重要而独特的作用。这些研究准备为组蛋白H2 A变体如何调节DDR和对IR的反应提供重要的见解。然后，我们将利用我们的初步分析，识别每个组蛋白H2 A变体的候选蛋白质相互作用因子，以确定单个组蛋白H2 A变体的DDR效应蛋白。最后，我们将测试我们的工作假设macroH 2A如何通过与我们已经确定的新DDR因子的相互作用促进HR的DNA修复。这些研究将为组蛋白H2 A变体如何塑造DDR以保护人类细胞中结构和功能多样的染色质景观的基因组完整性提供无与伦比的视角。DNA损伤剂是包括放射治疗在内的化疗剂的主要类别，并且表观基因组代表了药物发现的令人兴奋的新线索。因此，这项工作利用人类细胞中遗传、生化和细胞方法的组合来识别促进基因组和表观基因组完整性的关键DDR途径，以及对癌症相关疗法（包括放射和PARP抑制剂）的反应，这些疗法可以转化为改善人类癌症的治疗。