Epigenetic alterations after DNA damage repair drive treatment resistance in glioblastoma

DNA损伤修复后的表观遗传改变导致胶质母细胞瘤治疗耐药

• 批准号: 10525262
• 负责人: Aram Sandaldjian Modrek
• 金额: $ 14.17万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10525262
• 关键词: 3-Dimensional; Address; Binding; Biological Models; CRISPR/Cas technology; Cell Culture Techniques; Cells; ChIP-seq; Chemotherapy and/or radiation; Chromatin; Comet Assay; DNA; DNA Damage; DNA Methylation; DNA Modification Methylases; DNA Repair; DNA sequencing; DNA-Binding Proteins; Data; Enzymes; Epigenetic Process; Faculty; Fill-It; Gene Expression; Gene Expression Regulation; Genome; Glioblastoma; Goals; Guide RNA; HMGB2 Protein; Human; Individual; K-Series Research Career Programs; Knowledge; Lead; Life; Malignant Neoplasms; Malignant neoplasm of brain; Maps; Measures; Mediating; Mentors; Mission; Modeling; Molecular Analysis; Oncogenes; Patients; Phase; Population; Post-Translational Protein Processing; Principal Investigator; Process; Public Health; Radiation; Radiation Induced DNA Damage; Radiation induced damage; Radiation therapy; Radiation-Induced Change; Radiation-Sensitizing Agents; Radiosensitization; Recurrence; Regulatory Element; Research; Research Personnel; Resolution; Role; Site; Stress; Structure; System; Testing; Therapeutic; Training; United States National Institutes of Health; cancer cell; cancer therapy; career; career development; chemotherapy; chromosome conformation capture; cohesin; endonuclease; epigenome; experimental study; genome-wide; histone modification; in vivo; inhibitor; innovation; insight; meetings; novel therapeutics; recruit; repaired; response; skills; small molecule inhibitor; spatiotemporal; targeted treatment; therapeutic target; therapy resistant; time use; tool; transcription factor; transcriptome sequencing; treatment effect; tumor; tumor growth

PROJECT SUMMARY/ABSTRACT In this K08 career development award, the principal investigator (PI) aims to determine how DNA-methylation is altered after DNA damage and investigate the impact of these changes on 3D chromatin organization, gene regulation and treatment resistance. The PI studies this process in glioblastoma, a difficult to treat malignant brain tumor. Training and mentoring activities will facilitate meeting not only scientific goals, but also the principal investigator's career development by addressing gaps in knowledge and expanding training through coursework, meetings, networking, and an expert mentoring team consisting of a primary mentor, co-mentor, and four faculty advisors. The research proposed will be addressed through this K08 phase of training and serve as the scientific basis for the applicant's career as an independent investigator. The candidate will acquire skills necessary to complete the aims through selected mentors with non-overlapping expertise and coursework. The central hypothesis is that, after DNA damage repair, the local epigenetic state is not restored correctly, leading to epigenetic alterations, gene expression changes and treatment resistance. This hypothesis is tested using human patient-derived glioblastoma cell cultures as a model system. The rationale for this project is the observation that stochastic DNA methylation alterations can be detected with radiation damage models, and endonuclease damage can alter local DNA methylation states. The mechanism underlying this process and the extent to which it occurs in cancer, however, is not known. This hypothesis is challenging to test using stochastic damage, such as radiation, or traditional endonuclease damage models, which are unable to cut methylated DNA, and have a fixed and limited number of sites. To circumvent this issue, the investigator developed a CRISRP-Cas9 tool to reproducibly induce genome-wide double strand breaks to study DNA methylation alterations and genome organization around sites of DNA damage. The central hypothesis will be tested by two specific aims to (i) test how DNA methylation and genome organizational alterations evolve at damaged DNA loci, and (ii) test if genome re-organization factors can be targeted therapeutically during radiation stress. This training proposal is innovative because it (i) develops tools to map DNA methylation and 3D chromatin organization alterations following DNA damage and (ii) implicates this process in treatment resistance. The significance of this proposed research is that it fills knowledge gaps in epigenetics, DNA-damage repair, and the understanding of the effects of treatment on cancer cells. Successful completion of these studies will provide translatable insight into the interplay between DNA damage, DNA methylation and genome re-organization in glioblastoma.

项目总结/摘要 在这个K08职业发展奖，主要研究者（PI）的目的是确定DNA甲基化是如何 DNA损伤后改变，并研究这些变化对3D染色质组织，基因 调节和治疗阻力。PI在胶质母细胞瘤中研究了这一过程，胶质母细胞瘤是一种难以治疗的恶性肿瘤。 脑瘤培训和指导活动不仅有助于实现科学目标， 通过解决知识差距和通过课程扩大培训，促进调查员职业发展， 会议，网络，和一个专家指导团队组成的主要导师，共同导师，和四个教师 顾问。建议的研究将通过K08培训阶段进行，并作为科学的 申请人作为独立调查员的职业基础。候选人将获得必要的技能， 通过选定的具有非重叠专业知识和课程的导师完成目标。中央 假设是，在DNA损伤修复后，局部表观遗传状态没有正确恢复，导致 表观遗传改变、基因表达变化和治疗抗性。这个假设是用 人患者来源的胶质母细胞瘤细胞培养物作为模型系统。该项目的基本原理是 观察到随机DNA甲基化改变可以用辐射损伤模型检测， 核酸内切酶损伤可以改变局部DNA甲基化状态。这一过程背后的机制和 然而，它在癌症中发生的程度尚不清楚。这一假设是具有挑战性的测试使用随机 损伤，如辐射，或传统的核酸内切酶损伤模型，这是不能切割甲基化 DNA，并且具有固定且有限数量的位点。为了解决这个问题，研究人员开发了一个 CRISRP-Cas9工具可重复诱导全基因组双链断裂以研究DNA甲基化 改变和基因组组织周围的DNA损伤的网站。中心假设将由两个 具体目标是（i）测试DNA甲基化和基因组组织改变如何在受损DNA中进化 基因座，和（ii）测试基因组重组因子是否可以在辐射应激期间进行治疗靶向。这 培训提案是创新的，因为它（i）开发了绘制DNA甲基化和3D染色质的工具 组织改变后DNA损伤和（ii）牵连这一过程中的治疗阻力。的 这项研究的重要性在于它填补了表观遗传学，DNA损伤修复和遗传学方面的知识空白。 了解治疗对癌细胞的影响。成功完成这些研究将提供 DNA损伤，DNA甲基化和基因组重组之间的相互作用， 胶质母细胞瘤