Development of Mantle Cell Lymphoma Proliferation Signature Assay

项目摘要/摘要 这项名为“MCL35在套细胞淋巴瘤(MCL)中的检测进展”的提案是为了验证 基于数字基因表达的增殖信号量化的MCL预后分析 分析。MCL是一种B细胞恶性肿瘤，具有广泛的临床、病理和生物学特征。 MCL是由t(11；14)(q13；q32)易位定义的，它导致细胞周期蛋白D1蛋白的过度表达 和细胞周期失调。MCL患者有明显的不同临床行为，从懒惰到 高度侵袭性疾病，治疗选择从警惕等待到大剂量化疗 和干细胞移植。目前的治疗方法很大程度上是基于患者确诊时的年龄 而且目前还没有治疗标准。此前，我们的研究联盟，淋巴瘤和 白血病分子图谱计划(LLMPP)利用基因分析快速冷冻的MCL组织活检 表达谱分析(GEP)，并将“增殖特征”确定为最有效的相关生物标志物 与MCL患者的生存有关。然而，最初使用冷冻组织和GEP的技术是不切实际的。 用于常规诊断。一个潜在的简单解决方案是在组织活检上使用免疫组织化学来 评估Ki67抗原作为细胞增殖的全球标志物的存在。Ki67的研究已经被 在使用不同阈值的多项研究中预测预后；然而，重复性存在严重问题 在不同的实验室技术和病理学家的解释方法之间。因此，作为我们工作的一部分， 在美国国家癌症研究所的SPECSII计划中，我们开发了一种新的GEP增殖签名分析方法，用于 MCL在福尔马林固定的石蜡包埋组织(FFPET)中工作良好，称为“MCL35”。MCL35 采用临床级纳米串计数器系统，该系统已获得FDA批准，作为 ProSigna乳腺癌检测。MCL35精确度高、重复性好，实验室间重复性达到100% 在我们的初步工作中，使其成为临床诊断环境中应用的有力候选者。MCL35 自#年首次在美国临床肿瘤学会上口头介绍以来，该方法得到了关注。 2016年6月，我们正在与制药公司和临床试验合作团体讨论 申请化验。当前项目的目标是首先彻底分析验证 MCL35(UH2期)；然后，使用改进的分析方法对临床试验队列进行回顾性询问，以建立 临床有效(UH3期)。长期目标是使用产生的经过改进和验证的MCL35来 前瞻性地确定哪些MCL患者需要即时治疗意向治疗，以便设计 围绕这一高风险子集进行临床试验，并提高患者存活率。

PROJECT SUMMARY/ABSTRACT This proposal, “Assay Development of the MCL35 for Mantle Cell Lymphoma (MCL),” is written to validate a prognostic assay for MCL based on quantification of a proliferation signature using digital gene expression analysis. MCL is a B cell malignancy with a broad spectrum of clinical, pathological and biological features. MCL is defined by the t(11;14)(q13;q32) translocation, which results in over expression of the cyclin D1 protein and cell cycle dysregulation. MCL cases have markedly variable clinical behavior ranging from indolent to highly aggressive disease, with treatment options ranging from watchful waiting to high dose chemotherapy and stem cell transplant. The current approach to treatment is based largely on the patient's age at diagnosis and currently there is no therapeutic standard. Previously, our research consortium, the Lymphoma and Leukemia Molecular Profiling Project (LLMPP) analyzed snap frozen MCL tissue biopsies using gene expression profiling (GEP) and identified a “Proliferation Signature” as the most powerful biomarker correlating with patient survival in MCL. However, the original techniques using frozen tissues and GEP were impractical for routine diagnostics. A potentially easy solution has been to use immunohistochemistry on tissue biopsies to assess the presence of the Ki67 antigen as a global marker of cell proliferation. Ki67 studies have been prognostic in multiple studies using various thresholds; however, there are serious issues with reproducibility between different lab techniques and Pathologists' approach to interpretation. Therefore, as part of our work in the National Cancer Institute's SPECSII program, we developed a novel GEP proliferation signature assay for MCL that works well in formalin-fixed, paraffin-embedded tissues (FFPET) called the “MCL35”. The MCL35 uses the clinical-grade NanoString nCounter system, which is FDA-approved as the technical platform for the ProSigna Breast Cancer assay. The MCL35 is accurate, reproducible, with an inter-lab reproducibility of 100% in our initial work, making it a strong candidate for application in the clinical diagnostic setting. The MCL35 assay has gained attention since first being orally presented at the American Society of Clinical Oncology in June 2016, and we are in discussions with pharmaceutical companies and clinical trial cooperative groups on applications for the assay. The goals of the current project are first to thoroughly analytically validate the MCL35 (UH2 phase); then, use the refined assay to retrospectively interrogate clinical trial cohorts to establish it's clinical validity (UH3 phase). The long term goal is to use the resulting refined and validated MCL35 to prospectively identify those MCL patients in need of immediate curative intent therapy in order to design clinical trials around this high risk subset and improve patient survival.