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

项目摘要/摘要： 正常细胞的恶性转化是由关键细胞基因的致癌突变驱动的，其中 正在积极开发针对这些突变的治疗方法，以进行干预，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，有时会受到限制，具有挑战性，容易出现抽样错误 而且可能根本不可能。液体活检是获取肿瘤可操作突变的一个迅速崛起的领域。 基于循环肿瘤DNA(CtDNA)的体液中最小/非侵入性。 此UH2/UH3应用程序响应PAR-15-095以推进翻译和临床开发 EFIRM-液体活组织检查(ELB)是一种成熟的学术实验室分析方法，是对临床实验室的改进 加州大学洛杉矶分校分子诊断中的修正(CLIA)认证的实验室开发测试(ELB-LDT) 实验室(MDL)，CLIA认证/美国病理学家学会(CAP)认可的实验室。ELB- LDT将在临床背景下进行评估，用于询问非小细胞肺癌EGFR基因的可操作突变 退伍军人管理局大洛杉矶医疗系统(VA GLA)的患者。 将癌症检测、诊断和治疗的学术检测方法提高到规范的临床水平具有重要意义。 将为医生提供影响患者健康的新能力的分析。我们的建议是 先进的成熟的学术检测，EFIRM-液体活检(ELB)，提供最佳性能技术 为了检测致癌基因突变，成为CLIA认证的实验室开发检测(LDT)，EFIRM-Liquid 活检实验室开展试验(ELB-LDT)。ELB检测非小细胞肺癌患者中可操作的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。这个 临床验证研究将在加州大学洛杉矶分校医疗中心和退伍军人管理局大洛杉矶医疗中心进行 系统(VA GLA)，其中20%的腺癌患者亚型为非小细胞肺癌 (NSCLC)EGFR基因存在TKI反应突变。ELB-LDT技术如果经过验证，可能会取代 目前用于EGFR基因分型的液体活组织检查具有最好的性能，最小或非侵入性， 快速、廉价和独立的允许检测最常见的EGFR基因突变 可用TKI治疗，如吉非替尼或厄洛替尼，以有效延长肺无进展生存期 癌症患者。 实现这些目标有两个具体目标。目标1/UH2是为了适应学术EFIRM-Liquid 活组织检查(ELB)技术将成为加州大学洛杉矶分校分子病理学CLIA认证的检测方法 实验室，并确定分析和临床表现。目标2/UH3是分析和临床 在临床现场(加州大学洛杉矶分校和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).