Development of Mantle Cell Lymphoma Proliferation Signature Assay

项目概要/摘要 本提案“MCL 35用于套细胞淋巴瘤（MCL）的试验开发”旨在验证 基于使用数字基因表达的增殖标记的定量的MCL的预后测定 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由t（11;14）（q13;q32）易位定义，其导致细胞周期蛋白D1蛋白的过度表达 和细胞周期失调。MCL病例的临床表现有明显的变化，从懒惰到 高度侵袭性疾病，治疗选择范围从观察等待到高剂量化疗 和干细胞移植目前的治疗方法主要是根据患者的年龄诊断 目前尚无治疗标准。此前，我们的研究联盟，淋巴瘤和 白血病分子分析项目（LLMPP）使用基因分析技术分析了速冻MCL组织活检标本。 基因表达谱（GEP），并确定了“增殖签名”作为最强大的生物标志物， 与MCL患者生存率的关系。然而，使用冷冻组织和GEP的原始技术不切实际 进行常规诊断一个潜在的简单解决方案是在组织活检中使用免疫组织化学， 评估Ki 67抗原作为细胞增殖的总体标志物的存在。Ki 67研究已经被 在使用各种阈值的多项研究中预测;然而，再现性存在严重问题 不同的实验室技术和病理学家的解释方法之间的差异。作为我们工作的一部分， 通过美国国家癌症研究所的SPECSII计划，我们开发了一种新型GEP增殖特征检测方法， MCL在福尔马林固定的石蜡包埋组织（FFPET）中工作良好，称为“MCL 35”。MCL35 使用临床级NanoString nCounter系统，该系统经FDA批准为 ProSigna乳腺癌测定。MCL 35准确、重现性好，实验室间重现性为100% 在我们最初的工作中，使其成为临床诊断环境中应用的强有力候选者。MCL35 自年首次在美国临床肿瘤学会口头提出以来， 2016年6月，我们正在与制药公司和临床试验合作小组讨论 用于检测。当前项目的目标是首先彻底分析验证 MCL 35（UH 2期）;然后，使用改良试验回顾性询问临床试验队列，以确定 临床有效性（UH 3期）。长期目标是使用经过改进和验证的MCL 35， 前瞻性地确定那些需要立即治疗的MCL患者， 围绕这一高风险子集进行临床试验，并提高患者生存率。

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.