循环肿瘤DNA（ctDNA）突变的检测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，目前的 ctDNA 突变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分析前处理，这是一个耗时、昂贵且繁琐的过程，增加了样本污染的风险。为了克服这些限制，这里公开了 DNA/γPNA（γ 肽核酸）混合纳米报告基因的工程，用于通过原位分析和记录肿瘤特异性 DNA 突变进行 ctDNA 生物传感。 γPNA 对与 DNA 碱基配对中的单个错配的低耐受性使得能够高度选择性地识别和记录外周血中的 ctDNA 突变。由于其卓越的生物稳定性，由突变体 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。证明纳米报告仪对外周血ctDNA突变具有高度特异性，尿液分析清除的γPNA可以为肿瘤进展和预后评估提供有价值的信息。这项工作展示了纳米报告仪通过 ctDNA 突变的原位生物传感来监测尿液肿瘤和患者预后的潜力。© 2024 作者。 《Advanced Science》由 Wiley‐VCH GmbH 出版。

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.© 2024 The Author(s). Advanced Science published by Wiley‐VCH GmbH.