Abstract：
Detection of circulating tumor DNA (ctDNA) mutations, which are molecular biomarkers present in bodily fluids of cancer patients, can be applied for tumor diagnosis and prognosis monitoring. However, current profiling of ctDNA mutations relies primarily on polymerase chain reaction (PCR) and DNA sequencing and these techniques require preanalytical processing of blood samples, which are time-consuming, expensive, and tedious procedures that increase the risk of sample contamination. To overcome these limitations, here the engineering of a DNA/γPNA (gamma peptide nucleic acid) hybrid nanoreporter is disclosed for ctDNA biosensing via in situ profiling and recording of tumor-specific DNA mutations. The low tolerance of γPNA to single mismatch in base pairing with DNA allows highly selective recognition and recording of ctDNA mutations in peripheral blood. Owing to their remarkable biostability, the detached γPNA strands triggered by mutant ctDNA will be enriched in kidneys and cleared into urine for urinalysis. It is demonstrated that the nanoreporter has high specificity for ctDNA mutation in peripheral blood, and urinalysis of cleared γPNA can provide valuable information for tumor progression and prognosis evaluation. This work demonstrates the potential of the nanoreporter for urinary monitoring of tumor and patient prognosis through in situ biosensing of ctDNA mutations.
摘要：
检测循环肿瘤DNA（ctDNA）突变，它们是癌症患者体液中存在的分子生物标志物，可用于肿瘤诊断和预后监测。然而,目前ctDNA突变的分析主要依赖于聚合酶链反应（PCR）和DNA测序，这些技术需要对血液样本进行预分析处理，这很耗时，贵,和繁琐的程序，增加了样品污染的风险。为了克服这些限制,在此,公开了DNA/γPNA(γ肽核酸)杂合纳米报道分子的工程化,用于经由肿瘤特异性DNA突变的原位谱分析和记录的ctDNA生物传感。γPNA对与DNA碱基配对的单个错配的低耐受性允许高度选择性识别和记录外周血中的ctDNA突变。由于其显著的生物稳定性，由突变ctDNA触发的分离的γPNA链将在肾脏中富集并清除到尿液中进行尿液分析。结果表明，纳米报道分子对外周血中的ctDNA突变具有高特异性，尿液分析可以为肿瘤进展和预后评估提供有价值的信息。这项工作证明了纳米报道分子通过ctDNA突变的原位生物传感来监测肿瘤和患者预后的潜力。
