Quantification of cisplatin sensitivity and resistance using metabolic imaging and circulating tumor cell (CTC) biomarkers

项目3概要 顺铂（CDDP）仍然是多种实体瘤化疗治疗的金标准，包括 头颈部鳞状细胞癌。治疗失败率高是由于 在这种毒性相对较大的化疗后产生了获得性耐药性。尽管经常使用CDDP， 存在肿瘤反应或获得性抗性发展的预测因子。治疗失败一律是致命的。 鉴于对肿瘤反应和获得性耐药预测因子的关键未满足需求，我们将重点放在 利用微创成像（超极化磁共振成像）评估肿瘤反应的努力 共振成像; HP-MRI）和检测循环肿瘤细胞（CTC）中的生物学变化， 正在接受顺铂治疗我们已经证明，CDDP和其他遗传毒性剂触发 肿瘤细胞代谢的可测量波动，可通过常规生化测定和HP- 用[1 - 13C]-丙酮酸盐进行MRI。几种还原代谢途径中的关键辅酶的再循环， 基因毒性胁迫对丙酮酸通过乳酸脱氢酶（LDH）转化为乳酸的碳通量的影响。我们 先前表明，遗传毒性剂改变细胞内氧化还原状态，改变丙酮酸/乳酸代谢， 并以与抗肿瘤相关的方式抑制丙酮酸转化为乳酸（kPL）的表观速率 有效性基因组和转录组学分析与顺铂获得相关的调控变化 CTC中的耐药性将加强顺铂敏感性和耐药性的基于成像的定量。基于 根据这些初步数据，我们假设顺铂中Nrf-2驱动的代谢重编程- 使用碳通量的非侵入性成像（kPL-通过HP- MRI）和scCTC分析。为了评估这一前提，我们将使用充分表征的HNSCC临床前模型 对顺铂敏感和耐药的细胞将在基线时测量糖酵解代谢的变化， 通过超极化成像和生物化学测定观察顺铂给药后的变化。这些测量 将通过体外和体内生物化学试验进行验证（目标1）。我们还将测量治疗反应 在HNSCC患者中，通过在诱导前后采集HP-MRI数据， 化疗（目的2）。将在CTC中进行生物学确认，以鉴定与以下相关的生物标志物： 通过基因组和转录组学分析确定顺铂耐药性。成功完成本研究将 建立HP-MRI作为表征肿瘤对顺铂相对耐药性的微创成像方法 并提供实时反馈以优化治疗。CTC生物标志物分析将提供关键的生物学 支持成像结果。这项工作有可能改变临床决策的基础 关于顺铂在HNSCC和相关呼吸消化道癌症中的应用。作为值得注意的第一步 为了实现精确的肿瘤学方法，拟议的研究与NIH的使命有关， 它涉及开发和应用有助于减轻人类疾病负担的基础知识。

PROJECT 3 SUMMARY Cisplatin (CDDP) remains the gold-standard for chemotherapeutic treatment for multiple solid tumors, including head and neck squamous cell carcinoma (HNSCC). High rates of treatment failure result from the development of acquired resistance following this relatively toxic chemotherapy. Despite the frequent use of CDDP, no robust predictors of tumor response or development of acquired resistance exist. Treatment failure is uniformly fatal. Given the critical unmet need for predictors of tumor response and acquired resistance, we have focused our efforts on the assessment of tumor response with minimally invasive imaging (hyperpolarized magnetic resonance imaging; HP-MRI) and detection of biological shifts in circulating tumor cells (CTCs) while patients are undergoing cisplatin-based therapy. We have shown that CDDP and other genotoxic agents trigger measurable fluctuations in tumor cell metabolism detectable by both conventional biochemical assays and HP- MRI with [1-13C]-pyruvate. The recycling of key coenzymes in several reductive metabolic pathways links the effects of genotoxic stress to carbon flux from pyruvate into lactate via lactate dehydrogenase (LDH). We previously showed that genotoxic agents alter the intracellular redox state, shift pyruvate/lactate metabolism, and suppress the apparent rate of pyruvate conversion to lactate (kPL) in a manner that correlates with anti-tumor effectiveness. Genomic and transcriptomic analysis of regulatory shifts associated with the acquisition of cisplatin resistance in CTCs will re-enforce the imaging-based quantification of cisplatin sensitivity and resistance. Based on these preliminary data, we hypothesize that metabolic reprogramming driven by Nrf-2 in cisplatin- resistant HNSCC is detectable using a combination of non-invasive imaging of carbon flux (kPL- via HP- MRI) and scCTC analysis. To assess this premise, we will use well-characterized preclinical models of HNSCC that are sensitive and resistant to cisplatin. Alterations in glycolytic metabolism will be measured at baseline and following cisplatin administration through hyperpolarized imaging and biochemical assays. These measurements will be validated with biochemical assays in vitro and in vivo (Aim 1). We will also measure treatment response in HNSCC patients relative to alterations in tumor kPL by acquiring HP-MRI data prior to and following induction chemotherapy (Aim 2). Biological confirmation will be performed in CTCs to identify biomarkers associated with cisplatin resistance through genomic and transcriptomic analysis. Successful completion of this study will establish HP-MRI as a minimally invasive imaging approach to characterize relative tumor resistance to cisplatin and provide real-time feedback to optimize treatment. CTC biomarker analysis will provide critical biological support for the imaging findings. This work has the potential to change the basis for clinical decision-making regarding the use of cisplatin in HNSCC and related aerodigestive tract cancers. As a noteworthy first step towards a precision oncology approach, the proposed research is relevant to the part of the NIH’s mission that pertains to developing and applying fundamental knowledge that will help to reduce the burdens of human illness.