Blood-based monitoring of bladder-sparing trimodality therapy for muscle-invasive bladder cancer

项目总结/摘要 虽然根治性膀胱癌通常被推荐用于肌肉浸润性膀胱癌（MIBC），但耐受性良好的 另一种称为膀胱保留三联疗法（TMT），包括放化疗（CRT）， 经尿道膀胱肿瘤切除术（TURBT）与膀胱癌切除术具有相同的长期结局， 保留病人的膀胱然而，20-30%接受TMT治疗的患者需要补救性膀胱癌。 由于复发性疾病而切除，并且对分子生物标志物的需求未得到满足，以精确地识别适当的肿瘤。 并监测治疗后的复发情况。我们最近开发了微流感病毒- 从血液中分离循环肿瘤细胞（CTC）并对其进行分子表征， 作为非侵入性的、连续的“液体活组织检查”来预测和监测治疗反应。我们的长期目标是 开发预测性分子生物标志物，可以精确指导膀胱癌患者的个性化护理 癌本申请的总体目标是（i）优化用于肿瘤细胞的CTC和肿瘤分子特征， 预测TMT治疗患者的治疗反应和纵向监测，以及（ii）阐明 基于CTC和肿瘤分子谱的演变的治疗抗性的分子机制， CRT。我们的中心假设是，CTC分子特征与肿瘤分子特征相结合， 预测和监测MIBC患者膀胱保留治疗后的治疗反应。的理由 该项目的关键是，分子生物标志物的发展将使治疗的定制选择成为可能， 准确监测MIBC患者的治疗反应。中心假设将通过以下方式进行检验： 三个具体目标：1）鉴定与肿瘤分子标记物结合的治疗前CTC分子标记物， 与TMT后的预后相关; 2）鉴定CTC分子特征以监测早期检测 评价TMT治疗前后CTC和肿瘤分子谱的动态变化 TMT旨在阐明治疗耐药的机制。在第一个目标中，我们将优化预处理CTC， 接受TMT的MIBC患者的肿瘤预测特征，随后在入组的MIBC患者中进行验证 在SWOG/NRG 1806临床试验中。在第二个目标中，我们将优化用于早期检测的CTC RNA签名 在MIBC患者队列中TMT后复发率，随后在入组SWOG/NRG的患者中进行验证 1806.在第三个目标中，我们将分析从TMT纵向收集的CTC和肿瘤转录谱， 复发性疾病患者，以确定与治疗耐药性相关的分子特征。 在这项申请中提出的研究是创新的，因为它将开发基于 膀胱癌患者中CTC的微流体分离和RNA表达谱。拟议的重新- 搜索是重要的，因为它将产生临床上有用的生物标志物，用于精确选择患者， 保留膀胱的TMT和这些患者的非侵入性监测，同时也推进了我们的非侵入性监测。 了解膀胱癌对CRT反应和抵抗的分子决定因素。

PROJECT SUMMARY/ABSTRACT Although radical cystectomy is often recommended for muscle-invasive bladder cancer (MIBC), a well-tolerated alternative called bladder-sparing trimodality therapy (TMT), consisting of chemoradiation therapy (CRT) follow- ing transurethral resection of bladder tumor (TURBT), has equivalent long-term outcomes to cystectomy while preserving the patient’s native bladder. However, 20-30% of patients treated with TMT require a salvage cystec- tomy due to recurrent disease, and there is an unmet need for molecular biomarkers to precisely identify appro- priate candidates and to monitor for recurrences after therapy. We recently developed methods for the microflu- idic isolation and molecular characterization of circulating tumor cells (CTCs) from blood, which have potential as non-invasive, serial “liquid biopsies” to predict and monitor therapeutic responses. Our long-term goal is to develop predictive molecular biomarkers that can precisely guide the individualized care of patients with bladder cancer. The overall objectives of this application are to (i) optimize CTC and tumor molecular signatures for the prediction of therapeutic responses and longitudinal monitoring of patients treated with TMT, and (ii) to elucidate molecular mechanisms of treatment resistance based on the evolution of CTC and tumor molecular profiles after CRT. Our central hypothesis is that CTC molecular signatures, in combination with tumor molecular profiles, can predict and monitor therapeutic responses after bladder preservation therapy in patients with MIBC. The rationale for this project is that the development of molecular biomarkers will enable the tailored selection of therapy and accurate monitoring of treatment response for MIBC patients. The central hypothesis will be tested by pursuing three specific aims: 1) Identify pretreatment CTC molecular signatures that, combined with tumor molecular profiles, correlate with prognosis after TMT; 2) Identify CTC molecular signatures to monitor for early detection of recurrence after TMT; and 3) Evaluate dynamic changes in CTC and tumor molecular profiles before and after TMT to elucidate mechanisms of therapeutic resistance. In the first aim, we will optimize pretreatment CTC and tumor predictive signatures in MIBC patients undergoing TMT, followed by validation in MIBC patients enrolled in the SWOG/NRG 1806 clinical trial. In the second aim, we will optimize a CTC RNA signature for early detection of recurrence after TMT in a cohort of MIBC patients, followed by validation in patients enrolled in SWOG/NRG 1806. In the third aim, we will analyze CTC and tumor transcriptional profiles collected longitudinally from TMT patients who develop recurrent disease to identify molecular signatures associated with therapeutic resistance. The research proposed in this application is innovative because it will develop novel molecular assays based on the microfluidic isolation and RNA expression profiling of CTCs in bladder cancer patients. The proposed re- search is significant because it will yield clinically useful biomarkers for the precise selection of patients appro- priate for bladder-sparing TMT and the non-invasive monitoring of these patients, while also advancing our un- derstanding of molecular determinants of response and resistance to CRT in bladder cancer.