Liu Chang, Kannisto Eric, Yu Guan, Yang Yunchen, Reid Mary E, Patnaik Santosh K, Wu Yun
Department of Biomedical Engineering, University at Buffalo - The State University of New York, Buffalo, NY, United States.
Department of Thoracic Surgery, Roswell Park Comprehensive Cancer Center, Buffalo, NY, United States.
Front Genet. 2020 Mar 20;11:258. doi: 10.3389/fgene.2020.00258. eCollection 2020.
Circulating microRNAs carried by exosomes have emerged as promising diagnostic biomarkers for cancer because of their abundant amount and remarkable stability in body fluids. Exosomal microRNAs in blood are typically quantified using the RNA isolation-qRT-PCR workflow, which cannot distinguish circulating microRNAs secreted by cancer cells from those released by non-tumor cells, making it potentially less sensitive in detecting cancer-specific microRNA biomarkers. We have developed a sensitive and simple tethered cationic lipoplex nanoparticles (tCLN) biochip to detect exosomal microRNAs in human sera. The tCLN biochip allows the discrimination of tumor-derived exosomes from their non-tumor counterparts, and thus achieves higher detection sensitivity and specificity than qRT-PCR. We have demonstrated the clinical utility of the tCLN biochip in lung cancer diagnosis using sera from normal controls, therapy-naive early stage and late stage non-small cell lung cancer (NSCLC) patients. Total five microRNAs (miR-21, miR-25, miR-155, miR-210, and miR-486) were selected as the biomarkers. Each microRNA biomarker measured by tCLN assay showed higher sensitivity and specificity in lung cancer detection than that measured by qRT-PCR. When all five microRNAs were combined, the tCLN assay distinguished normal controls from all NSCLC patients with sensitivity of 0.969, specificity of 0.933 and AUC of 0.970, and provided much better diagnostic accuracy than qRT-PCR (sensitivity = 0.469, specificity = 1.000, AUC = 0.791). Remarkably, the tCLN assay achieved absolute sensitivity and specificity in discriminating early stage NSCLC patients from normal controls, demonstrating its great potential as a liquid biopsy assay for lung cancer early detection.
由于外泌体携带的循环微小RNA在体液中含量丰富且稳定性高，已成为很有前景的癌症诊断生物标志物。血液中的外泌体微小RNA通常采用RNA分离 - qRT - PCR流程进行定量，该方法无法区分癌细胞分泌的循环微小RNA和非肿瘤细胞释放的微小RNA，这使得其在检测癌症特异性微小RNA生物标志物时可能不太敏感。我们开发了一种灵敏且简单的 tethered阳离子脂质体纳米颗粒（tCLN）生物芯片来检测人血清中的外泌体微小RNA。tCLN生物芯片能够区分肿瘤来源的外泌体和非肿瘤来源的外泌体，因此比qRT - PCR具有更高的检测灵敏度和特异性。我们使用来自正常对照、未经治疗的早期和晚期非小细胞肺癌（NSCLC）患者的血清，证明了tCLN生物芯片在肺癌诊断中的临床应用价值。总共选择了5种微小RNA（miR - 21、miR - 25、miR - 155、miR - 210和miR - 486）作为生物标志物。通过tCLN检测法测量的每种微小RNA生物标志物在肺癌检测中都比通过qRT - PCR测量的显示出更高的灵敏度和特异性。当将所有5种微小RNA结合起来时，tCLN检测法能够区分正常对照和所有NSCLC患者，灵敏度为0.969，特异性为0.933，曲线下面积（AUC）为0.970，并且提供了比qRT - PCR更好的诊断准确性（灵敏度 = 0.469，特异性 = 1.000，AUC = 0.791）。值得注意的是，tCLN检测法在区分早期NSCLC患者和正常对照方面实现了绝对的灵敏度和特异性，证明了其作为肺癌早期检测液体活检检测方法的巨大潜力。
