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

基本信息

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
项目类别：
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

项目摘要

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.

抽象的目前尚无治愈与预后黯淡相关的高级别胶质瘤的方法和生活质量差。 GBM 患者通常接受手术切除，然后进行放射治疗疗法（RT）与烷化剂替莫唑胺（TMZ）相结合。然而，几乎所有 GBM 患者要么对这种治疗没有反应，要么最终产生耐药性，这强调了迫切需要有效的辅助治疗。 RT 和 TMZ 都可以通过引起肿瘤细胞致命的DNA双链断裂，可以被内在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 富含神经胶质瘤干细胞 (GSC) 的子集，其特征是 DDR 上调有助于其优越的治疗耐药性。靶向DNA修复可以增加/恢复脑肿瘤对 TMZ 和 RT 的敏感性，从而增加治疗效果疗效并最大限度地减少肿瘤复发。我们设计了一种快速、多功能且高度灵敏的报告基因，可检测和定量微量 DNA 双链断裂修复，例如同源定向修复以及非同源末端连接，无需破坏细胞。在此，我们建议使用该报告基因来识别 DNA 调节剂 GSC 中的损伤修复。在目标 1（R61 阶段）中，我们将开发和优化高通量监测 GSC 中 DDR 的筛选试验。在目标 2（R33 阶段）中，我们将使用此测定来筛选用于抑制 DNA 损伤修复的化合物。该项目完成后，此屏幕应鉴定出能够有效提高 GSC 对放化疗敏感性的新型化合物提高基因毒性治疗的功效。