Developing a CLIA compliant LDT for detecting plasma piR-Ls/pfeRNAs to distinguish benign and malignant lung nodules

开发符合 CLIA 标准的 LDT，用于检测血浆 piR-Ls/pfeRNA，以区分良性和恶性肺结节

基本信息

批准号：
9908056
负责人：
MALCOLM V BROCK
金额：
$ 33.16万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-05 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9908056
关键词：
Area Benign Biological Biological Assay Biological Markers Biopsy CLIA certified Cancer Etiology Cessation of life Clinical Clinical Markers Clinical Research Country Data Databases Decision Making Diagnosis Diagnostic Diagnostic Procedure Diagnostic tests Disease Early Diagnosis Enrollment Ensure Equipment Excision Future High Prevalence Histologic Hospitals In Vitro Laboratories Lung CAT Scan Lung nodule Malignant - descriptor Malignant Neoplasms Malignant neoplasm of lung Measures Medical Methods Non-Small-Cell Lung Carcinoma Nucleic Acids Patients Performance Physicians Plasma Play Reagent Reporting Research Personnel Role Sampling Scientist Sensitivity and Specificity Smoker Specimen Structure of parenchyma of lung Surgeon Survival Rate Technology Testing Time Tissues Validation X-Ray Computed Tomography base cohort deep sequencing diagnostic assay early detection biomarkers early phase clinical trial improved in vivo laboratory equipment lung cancer screening molecular diagnostics mortality multidisciplinary new technology next generation outcome forecast piRNA protein function response screening program tumorigenesis

项目摘要

Project Summary: Lung cancer is the leading cause of cancer-related death in the U.S., in large part, because the diagnosis tends to be made after the cancer has progressed to its most advanced stages. However, current diagnostic techniques for detecting early-stage non-small cell lung cancer (NSCLC) are either invasive or have poor accuracy. We recently reported that piRNA-like sncRNAs (piR-Ls) and protein function effector sncRNAs (pfeRNAs) play critical roles in the tumorigenesis and differentiation of lung cancer. We have developed a novel technology for accurately and robustly measuring piR-Ls and pfeRNAs in plasma. We also have identified 2 piR-Ls and 6 pfeRNAs as promising non-invasive biomarkers using next generation sncRNAs deep sequencing of 119 biospecimens, including (i) plasma from healthy controls (ii) patient plasma with the matched, corresponding histologically proven NSCLC tissue as well as histologically normal adjacent lung tissue from patients with Stage I/II NSCLC (iii) plasma from patients with both biopsy proven benign and malignant lung nodules. Furthermore, we validated these biomarkers in 352 clinical plasma specimens, including 77 healthy controls, 44 patients with benign disease as well as 231 patients with malignant lung nodules. Moreover, we formulated prediction rules for our assay in detecting early-stage NSCLC. Using only plasma, we were able to: (1) differentiate patients with and without lung nodules, with a sensitivity and specificity of 98.5% and 98.7%, respectively (an important clinical assay needed in remote, impoverished areas of the world with a high prevalence of smokers but no access to CT scanning); (2) differentiate patients who had malignant versus benign lung nodules, with a sensitivity and specificity of 96.5% and 72.7%, respectively (important for economically advanced countries enrolling smokers to CT lung cancer screening programs). These data strongly suggest that our panel of 2 piR-Ls and 6 pfeRNAs are non-invasive putative biomarkers for detecting early-stage lung cancers. Our multi-disciplinary collaborative team consists of a physician-scientist, basic researchers, a physician-scientist in the CLIA-certified laboratory, statisticians, and a commercial developer. This team will develop a CLIA compliant LDT for validating our nucleic- acid-based existing assay which now has been tested as valid in over 460 clinical specimens. The technologies that we will utilize for our testing are currently FDA-cleared for use in clinical laboratories, and the equipment as well as all reagents for our assays already are used in in-vitro-diagnostic testing. Our collaborative team aims to develop a molecular diagnostic assay that can be integrated into future clinical trials for the early detection of NSCLC, and ultimately, improve the dismal survival rates of that disease.

项目概要：肺癌是美国癌症相关死亡的主要原因，很大程度上是因为诊断倾向于在癌症进展到最晚期后进行。然而，目前的诊断技术用于检测早期非小细胞肺癌（NSCLC）的方法要么具有侵袭性，要么准确性较差。我们最近报道，piRNA 样 sncRNA (piR-Ls) 和蛋白质功能效应器 sncRNA (pfeRNA) 发挥作用在肺癌的发生和分化中发挥着重要作用。我们开发了一项新技术准确、稳健地测量血浆中的 piR-L 和 pfeRNA。我们还鉴定了 2 个 piR-L 和 6 个使用下一代 sncRNA 深度测序将 pfeRNA 作为有前途的非侵入性生物标志物 119 生物样本，包括 (i) 来自健康对照的血浆 (ii) 患者血浆以及匹配的相应血浆经组织学证实的 NSCLC 组织以及来自 2 期患者的组织学正常的邻近肺组织 I/II NSCLC (iii) 来自活检证实良性和恶性肺结节的患者的血浆。此外，我们在 352 份临床血浆样本中验证了这些生物标志物，其中包括 77 名健康对照者、44 名患有糖尿病的患者良性疾病以及 231 名恶性肺结节患者。此外，我们还制定了预测规则用于我们检测早期 NSCLC 的检测。仅使用血浆，我们能够：（1）区分患有以下疾病的患者：且无肺结节，敏感性和特异性分别为 98.5% 和 98.7%（一项重要的临床研究）世界上吸烟率高但又无法获得的偏远贫困地区需要进行检测 CT 扫描）； (2) 区分患有恶性和良性肺结节的患者，具有敏感性和特异性分别为 96.5% 和 72.7%（对于招募吸烟者的经济发达国家很重要） CT 肺癌筛查项目）。这些数据强烈表明我们的 2 个 piR-L 和 6 个 pfeRNA 组是用于检测早期肺癌的非侵入性假定生物标志物。我们的多学科协作团队由一名医师科学家、基础研究人员、一名 CLIA 认证实验室的医师科学家组成，统计学家和商业开发人员。该团队将开发符合 CLIA 标准的 LDT，用于验证我们的核酸基于酸的现有检测方法现已在 460 多个临床样本中进行了测试，证明其有效。技术我们将用于测试的设备目前已获得 FDA 批准可用于临床实验室，并且设备我们检测的所有试剂均已用于体外诊断测试。我们的协作团队旨在开发一种分子诊断检测方法，可以整合到未来的临床试验中，以早期检测非小细胞肺癌，最终提高了该疾病的低生存率。