Advancing EFIRM-Liquid Biopsy (eLB) to a CLIA-Certified Laboratory Developed Test (eLB-LDT) for Detection of Actionable EGFR Mutations in NSCLC Patients

项目概要/摘要： 正常细胞的恶性转化由关键细胞基因的致癌突变驱动， 目前正在积极开发针对这些突变的治疗方法，以改善发病率 和/或死亡率。目前的临床实践是通过肿瘤活检来询问这些“可操作的突变” 其次是基因分型，这是侵入性的，有时访问限制，具有挑战性，受抽样误差的影响 也许根本不可能液体活检是一个快速新兴的领域，以获得肿瘤中的可操作突变 基于循环肿瘤DNA（ctDNA）在体液中的最小/非侵入性。 该UH 2/UH 3应用响应PAR-15-095，以推进转化和临床开发 从一个成熟的学术实验室分析，EFIRM液体活检（eLB）到一个“临床实验室改进” 修正案”（CLIA）-认证的实验室开发的测试（eLB-LDT），在加州大学洛杉矶分校分子诊断 实验室（MDL），CLIA认证/美国病理学家学院（CAP）认证的实验室。eLB- LDT将在用于询问NSCLC EGFR基因中可操作突变的临床背景下进行评价 VA大洛杉矶医疗保健系统（VA GLA）的患者。 重要的是将用于癌症检测、诊断和治疗的学术测定推进到受监管的临床 这将为医生提供新的能力，以影响他们的患者的健康。我们的建议是 推进成熟的学术分析，EFIRM-液体活检（eLB），提供最佳的性能技术 检测致癌突变，成为CLIA认证的实验室开发测试（LDT），EFIRM液体 活检实验室开发的测试（eLB-LDT）。eLB检测NSCLC患者中可采取措施的EGFR突变， 与基于活检的基因分型100%一致，优于目前的液体活检技术。的 临床验证研究将在UCLA医学中心和VA Greater洛杉矶医疗中心进行 系统（VA GLA），其中20%的患者为腺癌亚型的非小细胞肺癌 （NSCLC）在EGFR基因中具有TKI应答突变。eLB-LDT技术如果得到验证， EGFR基因分型的液体活检的当前实践，因为它具有最佳性能，最小或非侵入性， 快速、廉价和独立，允许检测最常见的EGFR基因突变， 可以用TKI如吉非替尼或厄洛替尼治疗，以有效延长肺肿瘤的无进展生存期。 癌症患者。 为实现这些目标，有两个具体目标。目的1/UH 2是适应学术EFIRM-Liquid 活检（eLB）技术成为加州大学洛杉矶分校分子病理学诊断中CLIA认证的检测方法 实验室和确定分析和临床性能。目的2/UH 3是分析和临床 在临床研究中心（UCLA和VA GLA）验证eLB-LDT。

Project Summary/Abstract: Malignant transformation of normal cells are driven by oncogenic mutations of key cellular genes where therapeutics are actively being developed to target these mutations for interventions that will improve morbidity and/or mortality. The current clinical practice to interrogate these “actionable mutations” is by tumor biopsy followed by genotyping which is invasive and at times access restrictive, challenging, subject to sampling errors and may simply not possible. Liquid biopsy is a rapidly emerging field to access actionable mutations in tumors minimal/non-invasively in bodily fluids based on circulating tumor DNA (ctDNA). This UH2/UH3 application is responsive to the PAR-15-095 to advance the translational and clinical development of a matured academic laboratory assay, the EFIRM-Liquid Biopsy (eLB) to a “Clinical Laboratory Improvement Amendment” (CLIA)-certified laboratory developed test (eLB-LDT), in the UCLA Molecular Diagnostic Laboratories (MDL), a CLIA-certified/ College of American Pathologists (CAP)-accredited Laboratory. The eLB- LDT will be evaluated in a clinical context of use to interrogate actionable mutations in the EGFR gene of NSCLC patients at the VA Greater Los Angeles Healthcare System (VA GLA). It is important to advance academic assays for cancer detection, diagnosis and treatment to regulated clinical assays that will provide novel capabilities to physicians to impact health of their patients. Our proposal is to advance a matured academic assay, EFIRM-Liquid Biopsy (eLB) that delivers the best performance technology to detect oncogenic mutations, to become a CLIA-certified laboratory developed test (LDT), the EFIRM-Liquid Biopsy laboratory developed test (eLB-LDT). eLB detects actionable EGFR mutations in NSCLC patients with 100% concordance with biopsy-based genotyping, outperforms current technologies for liquid biopsy. The clinical validation study will be conducted at the UCLA Medical Center and VA Greater Los Angeles Healthcare System (VA GLA) where 20% of patients with adenocarcinoma subtype of non-small cell lung carcinoma (NSCLC) harbor TKI-responsive mutations in the EGFR gene. The eLB-LDT technology if validated, may replace current practice of liquid biopsy for EGFR genotyping as it has the best performance, minimal or non-invasive, rapid, inexpensive and self-contained permitting the detection of the most common EGFR gene mutations that are treatable with TKI such as Gefitinib or Erlotinib to effectively extend the progression free survival of lung cancer patients. Two Specific Aims are in place to achieve these goals. Aim 1/UH2 is to adapt the academic EFIRM-Liquid Biopsy (eLB) technology to become a CLIA-certified assay in the UCLA Diagnostics Molecular Pathology Laboratory and to determine analytical and clinical performance. Aim 2/UH3 is to analytically and clinically validate eLB-LDT at the clinical sites (UCLA and VA GLA).