Validation of blood-based predictive biomarkers of therapeutic response to neoadjuvant chemoradiation therapy in patients with locally advanced rectal cancer

项目总结/摘要 手术后的新辅助放化疗（nCRT）是以下患者的标准治疗： 局部晚期直肠癌（LARC），约占新诊断直肠癌病例的60%。 虽然nCRT使许多患者受益，但目前尚不清楚哪些患者会或不会产生有利反应。关于 20%接受nCRT的患者将出现病理学完全缓解（pCR），而约25%的患者将 没有反应。最近，一些临床试验已经评估了新的治疗模式，其结果 提出了新的治疗策略，这些策略以前对LARC患者是不可用的。的可用性 这些新的策略，至关重要的是能够通过预测性生物标志物先验地识别LARC患者， 对nCRT有反应或无反应的可能性更高。 我们的目标是优化和验证一种准确、精密和微创的高效检测方法 评估在LARC背景下预测nCRT缓解的生物标志物。待验证的检测试剂盒 在本提案中是使用xMAP技术的Luminex多重测定。我们已经确定了六个候选人 将在本研究中验证的生物标志物。在这里，使用优化和验证的检测，我们将实现 使用LARC患者在真实世界临床实验室环境中对候选生物标志物进行临床验证 标本在本研究结束时，我们将为患者实现微创、基于血液的检测 治疗分层，这将对LARC患者和医疗保健系统带来巨大益处。 使用xMAP检测试剂盒，我们正在检测DNA损伤反应（DDR）的候选生物标志物 来自LARC患者的冷冻保存的外周血单核细胞（PBMC）的细胞裂解物中的信号转导。在我们 这些生物标志物表达的变化与对 LARC患者的nCRT。这些候选生物标志物是通过我们最近的假设驱动的 由国防部资助的翻译研究。该提案是通过双方的合作制定的 主要调查员汇集了互补的专门知识。由多学科专家组成的团队 在临床和基础科学、检测开发/验证及其临床部署方面的专业知识。 在此，我们将分析验证我们的检测（UH 2阶段），然后通过使用 LARC患者（UH 3期）的生物样本。使用的生物样本来自LARC患者和健康人 Fox Chase生物样本库机构和LARC临床研究/试验的质控品，用于评估 治疗反应。在UH 2阶段，我们将进行预分析（生物样本变异性）和分析 (e.g.,精密度、准确度）验证。在UH 3阶段，我们将通过评价临床疗效来确定临床有效性 效用参数（例如，灵敏度/特异性、最佳截止值），并与nCRT应答状态进行比较。 成功完成将产生经验证的生物标志物/检测试剂，用作临床试验用检测试剂 试验/研究。从长远来看，这项工作可能会改善LARC患者治疗的患者分层。

PROJECT SUMMARY/ABSTRACT Neoadjuvant chemoradiation therapy (nCRT) followed by surgery is the standard of care for patients with locally advanced rectal cancer (LARC), which accounts for ~60% of newly diagnosed rectal cancer cases. Although nCRT benefits many patients, it is currently not known who will or will not respond favorably. About 20% patients that receive nCRT will experience a pathologic complete response (pCR), while ~25% patients will exhibit no response. Recently, several clinical trials have assessed new treatment paradigms, and their results present new treatment strategies which were previously not available for LARC patients. With the availability of these new strategies, it is crucial to be able to a priori identify LARC patients, via a predictive biomarker, who have a higher likelihood of responding or not responding to nCRT. Our goal is to optimize and validate an efficient assay that is accurate, precise, and minimally invasive to assess biomarker(s) that will predict response to nCRT in the setting of LARC. The assay that will be validated in this proposal is a Luminex multiplexed assay that uses the xMAP technology. We have defined six candidate biomarkers that will be validated in this study. Here, using the optimized and validated assay we will then achieve the clinical validation of the candidate biomarker(s) in a real-world clinical laboratory setting using LARC patient specimens. At the end of this study, we will achieve a minimally invasive, blood-based assay for patient treatment stratification which will be of tremendous benefit to LARC patient(s) and the healthcare system. Using the xMAP assay, we are detecting candidate biomarkers of DNA damage response (DDR) signaling in cell lysates from cryopreserved peripheral blood monocytes (PBMCs) of LARC patients. In our preliminary work, the change in the expression of these biomarkers significantly associated with response to nCRT in LARC patients. These candidate biomarkers were identified through our recent hypothesis-driven translational study funded by the DOD. This proposal was developed through a collaboration between the two primary investigators bringing together complementary expertise. The assembled multi-disciplinary team brings together expertise in clinical and basic science, assay development/validation, and their clinical deployment. Here, we will analytically validate our assay (UH2 phase) and then establish the clinical validity by using biospecimens from LARC patients (UH3 phase). The biospecimens used are from LARC patients and healthy controls from the Fox Chase BioSample Repository Facility, and from LARC clinical studies/trials evaluating therapeutic response. In the UH2 phase we will perform pre-analytical (biospecimen variability) and analytical (e.g., precision, accuracy) validation. In the UH3 phase, we will establish clinical validity by evaluating clinical utility parameters (e.g., sensitivity/specificity, optimal cut-offs) and compare with nCRT response status. Successful completion will yield validated biomarkers/assays for use as investigational assays in clinical trials/studies. Over the long-term, this work may improve patient stratification for treatment in LARC patients.