How HSP90 shapes genotype-phenotype relationships to alter treatment outcome in cancer

项目总结 癌症通过顺序获得突变而进化，这些突变共同扰乱了细胞的动态平衡。 从而造成了严重的破坏。这些错乱的细胞是如何存活的，形成肿瘤，积累额外的有害物质 突变并对治疗产生抵抗力？我建议用一本小说来解决这些重要的问题 基于蛋白质折叠原理的方法。我的工作证明了蛋白质折叠 伴侣热休克蛋白90(HSP90)减轻或“缓冲”人类的有害生物效应 突变。在这样做的过程中，HSP90改变了细胞对药物的敏感性，并影响了不同类型的 孟德尔病症。在这里，我假设通过缓冲突变，HSP90使肿瘤细胞能够阻止 癌症疗法。这项拟议的工作将确定HSP90如何以表观遗传学的方式改变 癌症中特定DNA修复突变的后果，以及HSP90缓冲如何影响肿瘤 对基因毒性疗法的反应。我将通过测量恶性疾病的影响来直接回答这些问题 转化自身对热休克蛋白90 S缓冲DNA修复蛋白突变的能力。我将决定是否有能力 HSP90用于缓冲Fanconi贫血(FA)DNA修复途径中一组特征良好的突变体，使用 通过转导已定义的致癌元件并使用遗传学方法在体外转化的细胞系 设计了FA患者来源的癌细胞系。作为这些研究在治疗上的补充，我 我还将研究HSP90在缓冲突变DNA修复蛋白中的作用以及HSP90如何帮助形成 肿瘤对基因毒性药物的反应。我将使用高通量的功能遗传学方法来解析 基于HSP90缓冲突变和非缓冲突变的临床癌症(由我最近定义 已发表和正在进行的工作)。结果将与后续的临床结果相关联 使用遗传毒性药物的标准护理治疗。这项K22奖将帮助我获得资金，以便 在领先机构的独立职位，建立独立研究的记录，并产生 我职业生涯R01阶段的初步数据。它还将为我提供癌症生物学和 以患者为导向的研究技能将帮助我竞争更广泛的资助机会。 我建议的工作将确定HSP90对突变积累的影响如何演变为 癌症的进展及其演变的作用如何使耐药性的出现。结果将提供一个 允许为患者识别和使用HSP90缓冲突变的理论框架 分层。这将有助于设计可能提供最大益处的个性化治疗， 包括以非常规方式使用现有的HSP90抑制剂。因此，这项工作将提供重要的 对肿瘤进化的基本机制的洞察以及开创一种改善 与多种癌症治疗相关的治疗干预措施的精确度和有效性。

PROJECT SUMMARY Cancers evolve through the sequential acquisition of mutations that collectively perturb cellular homeostasis thereby unleashing havoc. How do these deranged cells survive, form tumors, amass additional deleterious mutations and acquire resistance to therapies? I propose to tackle these important questions using a novel approach founded on principles of protein folding. My work has demonstrated that the protein-folding chaperone heat-shock protein 90 (HSP90) alleviates or “buffers” the deleterious biological effects of human mutations. In doing so, HSP90 alters cellular drug sensitivities and influences the clinical course of diverse Mendelian disorders. Here, I hypothesize that by buffering mutations, HSP90 enables tumor cells to thwart cancer therapies. The proposed work will determine how HSP90, acting epigenetically, alters the consequences of specific DNA repair mutations in cancers, and how HSP90 buffering influences tumor responses to genotoxic therapies. I will directly address these questions by measuring the effects of malignant transformation itself on HSP90's ability to buffer mutations in DNA repair proteins. I will determine the ability of HSP90 to buffer a panel of well-characterized mutants in the Fanconi Anemia (FA) DNA repair pathway, using cell lines transformed in vitro by transduction with defined oncogenic elements, and using genetically engineered FA patient-derived cancer cell lines. As a therapeutically relevant complement to these studies, I will also investigate the role of HSP90 in buffering mutant DNA repair proteins and how HSP90 helps shape tumor responses to genotoxic drugs. I will employ a high-throughput functional genetics approach to parse clinical cancers based on the presence of HSP90-buffered vs. non-buffered mutations (defined by my recently published and ongoing work) in DNA repair proteins. Results will be correlated with clinical outcome following standard-of-care treatment with genotoxic drugs. This K22 award will help me secure the funds for an independent position at a leading institution, establish a record of independent research, and generate preliminary data for the R01 phase of my career. It will also provide me with training in cancer biology and patient-oriented research skillsets that will help me compete for a broader spectrum of funding opportunities. The work I propose will determine how the influence of HSP90 on the accumulation of mutations evolves as cancers progress and how its evolving role enables the emergence of drug resistance. Results will provide a theoretical framework permitting the identification and use of HSP90-buffered mutations for patient stratification. This will contribute to the design of individualized treatments likely to provide the most benefit, including the use of existing HSP90 inhibitors in non-conventional ways. Thus, this work will provide important insights into fundamental mechanisms of tumor evolution as well as pioneer a strategy for improving the precision and efficacy of therapeutic interventions relevant to the treatment of many cancers.