Screening for adjuvant gliobalstoma therapeutics

胶质母细胞瘤辅助治疗的筛选

基本信息

批准号：
9127592
负责人：
BAKHOS A TANNOUS
金额：
$ 37.41万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
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：
2016-04-01 至 2019-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9127592
关键词：
Accounting Adjuvant Adjuvant Therapy Adult Alkylating Agents Apoptosis Astrocytes Biological Assay CCL4 gene Cell Survival Cells Central Nervous System Neoplasms Cessation of life Chemicals Cherry - dietary DNA DNA Adducts DNA Repair Data Analyses Diagnosis Dose Drug Targeting Drug resistance Enzymes Failure Fibroblasts Futile Cycling Genetic Genotype Glioblastoma Glioma Guanine Lead Lesion Libraries Luciferases Malignant - descriptor Malignant neoplasm of brain Mediating Methylation Molecular Newly Diagnosed Normal Cell Patients Pharmaceutical Chemistry Pharmaceutical Preparations Phase II Clinical Trials Phenotype Positioning Attribute Preclinical Drug Evaluation Primary Brain Neoplasms Progression-Free Survivals Radiation Radiosurgery Randomized Recurrence Recurrent tumor Reporter Resistance Source Specificity Stem cells System Testing Therapeutic Time Transferase Triage Update Validation analog base cell killing conventional therapy cytotoxicity drug candidate high throughput screening killings neoplastic cell novel novel therapeutics phase 3 study promoter public health relevance relating to nervous system research study response screening small molecule standard of care stem cell population stem cell therapy success temozolomide tumor tumor initiation

项目摘要

DESCRIPTION (provided by applicant): More than half of the 18,000 patients diagnosed with malignant primary brain tumors in the U.S. each year have glioblastoma (GBM), the most common and most aggressive primary malignant brain tumor in adults. Over the last two decades, the major breakthrough in the treatment for GBM has been the addition of the DNA alkylating agent temozolomide (TMZ) to the standard of care (surgery and radiation) yielding an increase in the median survival from 12.1 months to 14.6 months. Despite this advancement, 90% of GBM patients die within 5 years, a colossal failure attributed to TMZ resistance. One of the major predictor of GBM response to TMZ is the MGMT promoter methylation status. This enzyme removes the DNA adduct, induced by TMZ, leading to cell survival. Thus, patients whose tumors have transcriptional silencing of the MGMT gene, mediated by promoter methylation, are most likely to benefit from TMZ. Given that all glioblastomas recur to a tumor lesion with acquired resistance to TMZ, novel adjuvant therapies could be highly beneficial to GBM patients. Recently, it was shown that a subpopulation of tumor cells, called glioblastoma stem cells (GSCs or tumor initiating cells), are responsible for tumor recurrence and resistance to conventional therapies. In this proposal, we will use a multiplex secreted reporter system for high-throughput screening to find drugs that either synergizes with TMZ on GBM stem cells from both newly diagnosed and recurrent tumors, irrespective of the MGMT status, or that kills a specific GBM stem cells subtype. We will simultaneously screen in the same well for drugs which act on three different GBM stem cells subpopulations: (1) obtained from newly diagnosed tumors with methylated MGMT promoter; (2) newly diagnosed GBM with unmethylated MGMT promoter; (3) recurrent TMZ-resistant tumors. We will use primary GBM stem cells dissociated from patient tumors sections and grow them as neural spheres which maintain the phenotype/genotype of the original tumor. Dose- and time-dependent experiments of our drug hits will be performed and validated using secondary screening assays in culture. A set of analogues of the most promising hits will be purchased, if available commercially, or synthesized by medicinal chemistry to make these drugs "fit-for-purpose", to facilitate their study in cells, and for pull down assays to find targets of drug hits. Finally, specificity of drug hits o glioblastoma will be tested using a panel of normal cells and stem cells in culture.

描述（由申请人提供）：在美国，每年诊断出的 18,000 名恶性原发性脑肿瘤患者中，超过一半患有胶质母细胞瘤 (GBM)，这是过去二十年中成人中最常见和最具侵袭性的原发性恶性脑肿瘤。 GBM 治疗的重大突破是将 DNA 烷化剂替莫唑胺 (TMZ) 添加到护理标准（手术和放射）中尽管取得了这一进展，但 90% 的 GBM 患者会在 5 年内死亡，这是由于 TMZ 耐药而造成的巨大失败，GBM 对 TMZ 反应的主要预测因素之一是 MGMT 启动子。该酶可去除 TMZ 诱导的 DNA 加合物，从而导致肿瘤患者的 MGMT 基因转录沉默。鉴于所有胶质母细胞瘤都会复发并对 TMZ 产生耐药性，新型辅助疗法可能对 GBM 患者非常有益。胶质母细胞瘤干细胞（GSC 或肿瘤起始细胞）负责肿瘤复发和对常规疗法的耐药性。在本提案中，我们将使用多重分泌报告系统进行高通量筛选。发现无论 MGMT 状态如何，药物都可以与 TMZ 对来自新诊断和复发肿瘤的 GBM 干细胞产生协同作用，或者杀死特定的 GBM 干细胞亚型。我们将在同一孔中同时筛选作用于三种的药物。不同的GBM干细胞亚群：（1）从新诊断的具有甲基化MGMT启动子的肿瘤获得；（2）新诊断的具有非甲基化MGMT启动子的GBM；（3）复发性TMZ耐药的我们将使用从患者肿瘤切片中分离出来的原代 GBM 干细胞，并将其培养为维持原始肿瘤表型/基因型的神经球，我们将使用二次筛选进行和验证药物命中的剂量和时间依赖性实验。如果可以商业购买，将购买一组最有希望的命中的类似物，或通过药物化学合成，以使这些药物“适合目的”，以促进他们的研究。最后，将使用一组正常细胞和培养的干细胞来测试药物对胶质母细胞瘤的特异性。