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

项目总结： 在美国，肺癌是癌症相关死亡的主要原因，很大程度上是因为诊断往往 在癌症进展到最晚期后进行。然而，目前的诊断技术 对于早期非小细胞肺癌(NSCLC)的检测，要么是侵袭性的，要么是准确性差的。我们 最近报道了piRNA样单链RNA(Pir-L)和蛋白功能效应子单链单链RNA(PfeRNAs)的作用 在肺癌发生和分化中的关键作用。我们已经开发了一种新的技术来 准确可靠地检测血浆中的Pir-L和pFeRNAs。我们还鉴定了2个Pir-L和6个Pir-L 利用新一代SncRNAs深度测序119作为有前途的非侵入性生物标志物 生物标本，包括(I)来自健康对照的血浆(Ii)患者血浆与相匹配的、相应的 经组织学证实的非小细胞肺癌组织及正常肺组织 经活检证实为良性和恶性肺结节患者的I/II NSCLC(III)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，44名慢性阻塞性肺病患者 良性疾病以及231例恶性肺结节患者。此外，我们还制定了预测规则 用于我们检测早期非小细胞肺癌的方法。仅使用血浆，我们就能够：(1)区分患有 无肺结节，其敏感性和特异性分别为98.5%和98.7%(临床意义重大 世界上吸烟者居高不下但无法获得 CT扫描)；(2)鉴别肺结节的良恶性。 分别为96.5%和72.7%的特异性(对于经济发达国家招募吸烟者来说很重要 到CT肺癌筛查计划)。这些数据强烈表明，我们的2个Pir-L和6个pfeRNA组成的小组 是检测早期肺癌的非侵入性生物标志物。我们的多学科协作 团队由一名内科科学家、基础研究人员、CLIA认证实验室的一名内科科学家、 统计学家和一个商业开发商。这个团队将开发一种符合CLIA的LDT，用于验证我们的核- 现有的以酸为基础的检测方法现已在460多个临床样本中被测试为有效。这些技术 我们将用于我们的测试的设备目前已获得FDA批准，可以在临床实验室使用，并且设备 我们检测的所有试剂都已用于体外诊断测试。我们的协作团队旨在 开发一种分子诊断方法，可整合到未来的临床试验中，用于早期检测 NSCLC，并最终提高这种疾病令人沮丧的存活率。

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.