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.

项目摘要 对烷基化剂的抗性，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。一个重要的区别 这些疗法是单官能烷基形成几种类型的烷基化核苷酸，而双功能 烷基产物形成剧毒链间交联（ICL）。我们假设CCNU和TMZ杀死神经胶质瘤 通过不同的机制进行细胞，在某些情况下导致不同的电阻模式和 对结果的独立影响可能是为了治疗益处的利用。对两者的抵抗力 药物可能是由甲酯甲基转移酶（MGMT）蛋白的表达产生的。 TMZ抗性 也可能是由于不匹配修复缺陷（MMR-D）引起的。对CCNU的抵抗力尚未系统地 表征但涉及几种DNA损伤修复（DDR）途径的成员。我们将扩展以前 通过系统地探测DDR途径的工作 基因组和功能基因组技术检验以下假设： AIM 1：通过MMR-D通过MMR-D抗性预测对CCNU的敏感性。我们最近的工作表明MMR-D 不会引起CCNU抗性。使用等源性和患者衍生的模型，我们将确定哪种TMZ 耐药机制还导致CCNU的抗性，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测定法来检验 在某些情况下，TMZ和CCNU的组合可以比单一疗法提供治疗益处。 AIM 2：维修蛋白质与CCNU诱导的ICL相关的蛋白质确定对CCNU的敏感性。我们将测试 已知有助于ICL修复CCNU抗性的基因转录激活的影响。然后我们会 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 神经胶质瘤前后CCNU治疗的基因组表征，以表征突变和SV特征 CCNU耐药性以及CCNU治疗对基因组不稳定性的影响。该提议将脱离 可能在临床上可能是对单官能烷基化剂的耐药性差异 可利用神经胶质瘤患者改善预后。 Broad Institute和Dana Farber癌症研究所的培训将为Simona Dalin博士提供技能 教师，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。她还将指导学生在项目的各个方面。