Screening for DNA damage response modulators in glioblastoma stem cells

胶质母细胞瘤干细胞中 DNA 损伤反应调节剂的筛选

• 批准号: 10038588
• 负责人: Christian Elias Badr
• 金额: $ 42万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10038588
• 关键词: Acidosis; Adjuvant; Adjuvant Therapy; Alkylating Agents; Alternative Therapies; Biological Assay; Biological Process; Blood; Brain Neoplasms; Cell Death; Cell Survival; Cells; Cellular Stress; Chemotherapy and/or radiation; Conditioned Culture Media; DNA; DNA Damage; DNA Double Strand Break; DNA Repair; DNA Sequence Analysis; Dose; Double Strand Break Repair; Drug Screening; Drug Targeting; Environment; Enzymes; Event; Excision; Experimental Animal Model; Fatty Acid Desaturases; Genetic; Glioblastoma; Glioma; Hypoxia; Impairment; Left; Libraries; Luciferases; Malignant - descriptor; Malignant Neoplasms; Measures; Mediating; Molecular Profiling; Monitor; Nonhomologous DNA End Joining; Normal Cell; Nutrient; Operative Surgical Procedures; Pathway interactions; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacology; Phase; Population; Primary Brain Neoplasms; Proliferating; Proteins; Quality of life; Rad51 recombinase; Radiation therapy; Recurrence; Reporter; Reporting; Resistance; Resistance development; System; Testing; Therapeutic; Time; Treatment Efficacy; analog; base; cancer stem cell; chemoradiation; conventional therapy; design; genotoxicity; high throughput screening; inhibitor/antagonist; neoplastic cell; novel; outcome forecast; oxidative DNA damage; patient variability; radioresistant; repaired; response; screening; small molecule; stem cells; stem-like cell; temozolomide; therapy resistant; tumor; virtual

Abstract There is presently no cure for high grade gliomas which are associated with a bleak prognosis and a poor quality of life. GBM patients typically undergo surgical resection followed by radiation therapy (RT) combined with the alkylating agent temozolomide (TMZ). However, virtually all GBM patients either fail to respond to this treatment, or ultimately develop resistance, underlining the urgency for effective adjuvant therapies. Both RT and TMZ can target tumor cells by causing lethal DNA double-strand breaks, which could be recognized and repaired by the intrinsic DNA damage response (DDR) thus protecting against DNA damage-induced cell death. Further, GBMs are enriched in a subset population of glioma cancer stem cells (GSCs), characterized by an upregulated DDR which contributes to their superior therapeutic resistance. Targeting DNA repair can increase/restore sensitivity of brain tumors to TMZ and RT, thus increasing therapeutic efficacy and minimizing tumor recurrence. We have designed a rapid, versatile and highly sensitive reporter that can detect and quantify minute amounts of DNA double-strand break repairs such as homology-directed repair as well as non-homologous end joining without disrupting cells. Here we propose to use this reporter in order to identify modulators of DNA damage repair in GSCs. In Aim 1 (R61 phase), we will develop and optimize a high-throughput screening assay to monitor DDR in GSCs. In Aim 2 (R33 phase), we will use this assay to screen for compounds which inhibit DNA damage repair. Upon completion of this project, this screen should identify novel compounds which effectively sensitize GSCs to chemoradiotherapy and increase the efficacy of genotoxic therapeutics.

抽象的 目前无法治愈高级神经胶质瘤，与预后相关 和生活质量差。 GBM患者通常接受手术切除，然后进行辐射 疗法（RT）与烷基化剂替莫唑胺（TMZ）结合使用。但是，几乎所有GBM 患者要么无法对此治疗做出反应，要么最终产生抗药性，强调 有效辅助疗法的紧迫性。 RT和TMZ均可通过引起肿瘤细胞 致命的DNA双链断裂，可以通过内在的DNA识别和修复 损伤反应（DDR），从而防止DNA损伤引起的细胞死亡。此外，GBMS 富含胶质瘤癌干细胞（GSC）的子群，其特征是 上调的DDR有助于其出色的治疗性。靶向DNA修复 可以增加/恢复脑肿瘤对TMZ和RT的敏感性，从而增加治疗性 功效和最小化肿瘤复发。我们设计了一个快速，通用和高度的 敏感的记者可以检测和量化微小量的DNA双链断裂 诸如同源指导的维修以及非同源结局之类的维修无需 破坏细胞。在这里，我们建议使用此记者以识别DNA的调节剂 GSC的损坏维修。在AIM 1（R61阶段）中，我们将开发和优化高通量 筛选分析以监视GSC中的DDR。在AIM 2（R33阶段）中，我们将使用此测定法进行筛选 对于抑制DNA损伤修复的化合物。该项目完成后，此屏幕 应鉴定出有效敏感GSC对化学放射疗法的新型化合物和 增加遗传毒性疗法的功效。