Differential resistance mechanisms to monofunctional vs bifunctional alkylating agents in glioma

神经胶质瘤对单功能烷化剂与双功能烷化剂的不同耐药机制

• 批准号: 10374792
• 负责人: Simona Dalin
• 金额: $ 6.76万
• 依托单位: BROAD INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2024-07-10
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10374792
• 关键词: Alkylating Agents; Alkylation; BRCA mutations; Bar Codes; Biological Assay; Biology; Carmustine; Cells; Characteristics; Chemicals; Chemoresistance; Clinical; Clinical Trials; Computer Analysis; DNA Damage; DNA Interstrand Crosslinking; DNA Probes; DNA Repair; Dana-Farber Cancer Institute; Data; Evaluation; Faculty; Gene Activation; Genomic Instability; Genomics; Glioma; Guanine; Human; Institutes; Knowledge; Lesion; Light; Lomustine; Malignant Glioma; Mediating; Methylation; Mismatch Repair; Mismatch Repair Deficiency; Modeling; Modernization; Molecular; Mutation; Nitrosourea Compounds; Nucleotides; Outcome; Pathway interactions; Patient-Focused Outcomes; Patients; Pattern; Pharmaceutical Preparations; Phenotype; Prognosis; Proteins; Radiation; Regimen; Resistance; Techniques; Technology; Testing; Therapeutic; Toxic effect; Training; Transcriptional Activation; Transferase; Ursidae Family; Variant; Work; adduct; algorithm development; biomarker-driven; chemotherapeutic agent; clinical biomarkers; clinical implementation; crosslink; design; detection method; effective therapy; functional genomics; genomic data; genomic signature; improved; improved outcome; inhibitor; member; patient population; phase III trial; prevent; promoter; rational design; repaired; resistance mechanism; response biomarker; skills; student mentoring; targeted treatment; temozolomide; therapy development; treatment optimization; treatment strategy; tumor

Project Summary Resistance to alkylating agents, including monofunctional temozolomide (TMZ) and bifunctional CCNU, remains a major obstacle to improving outcomes for patients with glioma. An important difference between these therapies is that monofunctional alkylators form several types of alkylated nucleotides, while bifunctional alkylators form highly toxic interstrand crosslinks (ICLs). We hypothesize that CCNU and TMZ kill glioma cells through different mechanisms, leading in some instances to different patterns of resistance and independent effects on outcome that may be leveraged for therapeutic benefit. Resistance to both agents can result from expression of the methyl guanine methyl transferase (MGMT) protein. TMZ resistance can also result from mismatch repair deficiency (MMR-d). Resistance to CCNU has not been systematically characterized but involves members of several DNA damage repair (DDR) pathways. We will extend previous work by systematically probing DDR pathways using varied and representative models and state-of-the-art genomic and functional genomic technologies to test the following hypotheses: Aim 1: TMZ resistance via MMR-d predicts sensitivity to CCNU. Our recent work suggests that MMR-d does not cause CCNU resistance. Using isogenic and patient derived models, we will determine which TMZ resistance mechanisms also result in resistance to CCNU and assess changes in mutational signatures after CCNU treatment. We will then use competition assays and barcoding assays to test the hypothesis that the combination of TMZ and CCNU may provide therapeutic benefit over monotherapy in some contexts. Aim 2: Repair proteins that engage CCNU-induced ICLs determine sensitivity to CCNU. We will test the effect of transcriptional activation of genes known to contribute to ICL repair on CCNU resistance. We will then determine if chemical inhibitors of those proteins can improve CCNU efficacy. Finally, we will perform complete genomic characterization of gliomas pre- and post- CCNU treatment to characterize mutational and SV profiles of CCNU resistance as well as the effect of CCNU treatment on genomic instability. This proposal will shed light on differences in resistance to mono- and bi-functional alkylating agents that may be clinically exploitable to improve outcomes for patients with glioma. This training at the Broad Institute and Dana Farber Cancer Institute will provide Dr. Simona Dalin skills of a faculty member, including systematic and genomic techniques, computational analysis of genomic datasets, algorithm development, and DDR biology. She will also mentor students in aspects of the project.

项目概要 对烷化剂的耐药性，包括单功能替莫唑胺 (TMZ) 和双功能 CCNU， 仍然是改善神经胶质瘤患者预后的主要障碍。之间的一个重要区别 这些疗法的特点是单功能烷基化剂形成几种类型的烷基化核苷酸，而双功能烷基化剂形成多种类型的烷基化核苷酸，而双功能烷基化剂形成多种类型的烷基化核苷酸。 烷基化剂形成剧毒的链间交联（ICL）。我们假设 CCNU 和 TMZ 可以杀死神经胶质瘤 细胞通过不同的机制，在某些情况下导致不同的抵抗模式和 对可用于治疗益处的结果的独立影响。对两者都有抵抗力 药物可以由甲基鸟嘌呤甲基转移酶（MGMT）蛋白的表达产生。替莫唑胺耐药 也可能由错配修复缺陷 (MMR-d) 引起。对 CCNU 的抵制尚未系统化 具有特征，但涉及多种 DNA 损伤修复 (DDR) 途径的成员。我们将延续之前的 通过使用各种代表性模型和最先进的技术系统地探索 DDR 路径来开展工作 基因组和功能基因组技术来检验以下假设： 目标 1：通过 MMR-d 的 TMZ 耐药性预测对 CCNU 的敏感性。我们最近的工作表明 MMR-d 不会引起 CCNU 耐药。使用同基因模型和患者衍生模型，我们将确定哪种 TMZ 耐药机制也会导致对 CCNU 的耐药性，并评估突变特征的变化 CCNU 治疗。然后，我们将使用竞争分析和条形码分析来检验以下假设： 在某些情况下，TMZ 和 CCNU 的组合可能比单一疗法提供治疗益处。 目标 2：与 CCNU 诱导的 ICL 结合的修复蛋白决定对 CCNU 的敏感性。我们将测试 已知有助于 ICL 修复的基因转录激活对 CCNU 抗性的影响。我们随后将 确定这些蛋白质的化学抑制剂是否可以提高 CCNU 功效。最后我们将执行完整的 CCNU 治疗前后神经胶质瘤的基因组特征，以表征突变和 SV 谱 CCNU 耐药性以及 CCNU 治疗对基因组不稳定性的影响。该提案将流 阐明对单功能烷化剂和双功能烷化剂的耐药性差异可能在临床上 可用于改善神经胶质瘤患者的预后。 布罗德研究所和达纳法伯癌症研究所的培训将为 Simona Dalin 博士提供以下技能： 教职人员，包括系统和基因组技术、基因组数据集的计算分析、 算法开发和 DDR 生物学。她还将在该项目的各个方面指导学生。