Extracellular Vesicles in Small Cell Lung Cancer Early Detection

项目摘要 肺癌是全球癌症死亡的主要原因。在实施肺癌 非小细胞肺癌（NSCLC）亚型的筛查为这种疾病带来了重大希望， 小细胞肺癌（SCLC）的早期检测选择有限，SCLC的5年生存率 只有7%，尽管开发了新的靶向治疗和NSCLC的早期检测， SCLC已取得进展。我们目前肺癌检测和治疗方法的差距 用于检测和监测肺癌的信息量大且可靠的生物标志物， 仍然难以捉摸微RNA（miRNAs）已经成为体液中可行的生物标志物，因此，提供了一种新的生物标志物。 这是实现早期癌症检测的非侵入性测定的极好手段。此外，miRNA表达在 循环似乎是隔室特异性的。虽然大多数miRNAs是细胞内的，但一个显著的 已经在细胞外观察到许多miRNA，包括在各种体液中。起源， RNA在循环中的应用和潜在功能是有趣问题的来源。获得 血浆中详细的RNA谱将有助于阐明这一问题。我们进行了多学科的 研究循环RNA转录本的方法，整合了miRNA生物学的专业知识， 纳米工程、肺癌和生物信息学。我们开发了一种简单的拴系阳离子脂质复合物 在Lipoplex纳米颗粒中预加载分子信标（MB）作为探针的纳米颗粒（tCLN）生物芯片 捕获和检测人血浆中的靶向miRNA和mRNA，而不需要任何预采样或后采样 治疗我们已经成功地证明了使用基因芯片评估外泌体miRNAs和mRNAs的能力。 下一代测序和我们的tCLN生物芯片在对照吸烟者和早期吸烟患者的队列中， 晚期NSCLC。我们的主要目标是通过（1）测试和验证 在一项回顾性和前瞻性研究中，测量ASCL 1和DLL 3在SCLC早期检测中的作用， 网络样本（2）使用巢代技术开发一组综合EV RNA候选物 测序和q-RT-PCR（3）建立优化的基于EV纳米芯片的早期小细胞肺癌RNA分类器 （4）利用多重TLN阵列生物芯片对优化后的EV RNA分类器进行检测， OSU James癌症医院和SCLC联盟的独立、盲法病例对照研究。

PROJECT SUMMARY Lung cancer is the leading cause of cancer deaths worldwide. While the implementation of lung cancer screening for non-small cell lung cancer (NSCLC) subtypes has brought significant hope to this disease, very limited options exist for the early detection of small cell lung cancer (SCLC) SCLC carries a 5-year survival rate of only 7% and despite the development of novel targeted therapies and early detection for NSCLC, no such advances have been achieved in SCLC. A gap in our current approach to lung cancer detection and treatment has been that informative and reliable biomarkers for the detection and surveillance of lung cancer have remained elusive. MicroRNAs (miRNAs) have emerged as viable biomarkers in body fluids thus, providing an excellent means to achieve non-invasive assays for early cancer detection. Furthermore, miRNA expression in circulation appears to be compartment specific. While the majority of miRNAs are intracellular, a significant number of miRNAs have been observed outside of cells, including in various bodily fluids. The origin, applications and potential functionality of RNAs in circulation are the sources of intriguing questions. Obtaining a detailed RNA spectrum in plasma would shed some light on this matter. We have taken a multidisciplinary approach to the investigation of circulating RNA transcripts that integrates expertise in miRNA biology, nanoengineering, lung cancer and bioinformatics. We have developed a simple tethered Cationic Lipoplex Nanoparticle (tCLN) biochip with pre-loaded molecular beacons (MBs) in the lipoplex nanoparticles as probes to capture and detect targeted miRNAs and mRNAs in human plasma without any need of pre- or post-sample treatment. We have successfully demonstrated the ability to assess both exosomal miRNAs and mRNAs using both Next Generation Sequencing and our tCLN biochip in cohorts of control smokers and patients with early stage NSCLC. Our primary objectives are to extend these novel findings by (1) Test and validate the utility of measurement of ASCL1 and DLL3 in the early detection of SCLC in a retrospective and prospective study with network samples (2) Develop A Panel of Comprehensive EV RNA Candidates using nest generation sequencing and q-RT-PCR (3) Develop an optimized EV nanochip based RNA Classifier for early SCLC detection and (4) Validate the optimized EV RNA Classifier by using the multiplex TLN array biochips in independent, blinded case control studies at the OSU James Cancer Hospital and from the SCLC consortium.