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: 10683338
• 负责人: Christian Elias Badr
• 金额: $ 32.26万
• 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 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10683338
• 关键词: Acidosis; Adjuvant; Adjuvant Therapy; Alkylating Agents; Alternative Therapies; Biological Assay; Biological Process; Blood; Brain Neoplasms; Cell Death Induction; Cell Survival; Cells; Cellular Stress; Cessation of life; Chemotherapy and/or radiation; Combined Modality Therapy; Conditioned Culture Media; 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; Operative Surgical Procedures; Pathway interactions; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phase; Population; Primary Brain Neoplasms; Prognosis; Proliferating; Proteins; Quality of life; Rad51 recombinase; Radiation therapy; Recurrence; Recurrent tumor; Reporter; Reporting; Resistance; Resistance development; System; Testing; Therapeutic; Time; Treatment Efficacy; analog; cancer stem cell; chemoradiation; conventional therapy; counterscreen; design; effective therapy; genotoxicity; high throughput screening; inhibitor; multiplex assay; neoplastic cell; novel; nutrient deprivation; oxidative DNA damage; patient variability; pharmacologic; 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.

摘要 目前还没有治愈高级别胶质瘤的方法，高级别胶质瘤的预后很差 生活质量很差GBM患者通常接受手术切除，然后进行放射治疗 治疗（RT）与烷化剂替莫唑胺（TMZ）的组合。然而，几乎所有的GBM 患者要么对这种治疗没有反应，要么最终产生耐药性， 迫切需要有效的辅助治疗。RT和TMZ都可以靶向肿瘤细胞， 致死性DNA双链断裂，可被内源性DNA识别和修复 损伤反应（DDR），从而防止DNA损伤诱导的细胞死亡。此外，GBM 在神经胶质瘤癌干细胞（GSC）的亚群中富集，其特征在于， 上调的DDR，这有助于它们的上级治疗抗性。靶向DNA修复 可以增加/恢复脑肿瘤对TMZ和RT的敏感性，从而增加治疗效果。 有效性和最小化肿瘤复发。我们设计了一种快速、多功能、 一种灵敏的报告基因，可以检测和定量微量的DNA双链断裂 修复如同源定向修复以及非同源末端连接， 破坏细胞。在这里，我们建议使用这种报告，以确定DNA的调节剂 GSC中的损伤修复。在目标1（R61阶段），我们将开发和优化高通量 筛选测定以监测GSC中的DDR。在Aim 2（R33阶段），我们将使用该试验筛选 寻找抑制DNA损伤修复的化合物在这个项目完成后，这个屏幕 应该鉴定有效地使GSC对放化疗敏感的新化合物， 增加遗传毒性治疗的功效。