Precision blood biopsy for lung cancer diagnosis using a targeting nanoprobe to detect multiple mRNA biomarkers in circulating malignant cells

Compared to tissue biopsy, blood biopsy offers significant advantages in terms of safety and convenience. However, achieving accurate blood biopsy for cancer pathological diagnosis presents substantial challenges. Herein, we have developed a precision blood biopsy technology utilizing a malignant cell-targeted nanoprobe for lung cancer diagnosis. The nanoprobe functionalized with SYL3C-conjugated hyaluronic acid and the KALA-GE11 peptide can efficiently deliver the loaded molecular beacons into heterogeneous circulating malignant cells (CMCs) in whole blood to detect diverse intracellular mRNA markers, including the proliferation marker Ki67, the oncogenic marker c-Myc, and thyroid transcription factor 1 (TTF-1) typically expressed in specific types of lung cancers. The simultaneous detection of various markers not only reduces false-negative rates caused by the high heterogeneity of CMCs but also provides accurate insight into the specific heterogeneity of CMCs for individual patients. The nanoprobe can differentiate lung adenocarcinoma (LAD) and small cell lung cancer (SCLC) from squamous cell carcinoma (SCC). TTF-1 is abundant in CMCs of LAD and SCLC but is relatively less common in SCC. c-Myc is more frequently overexpressed in CMCs from LAD and SCLC. Notably, Ki67 is upregulated in most CMCs across all lung cancer types. Furthermore, this approach can sensitively identify malignant cells from early stage lesions clinically classified as noninvasive carcinoma in situ. This technique holds significant clinical potentials for the early detection and precise characterization of lung cancer. Importantly, this strategy is adaptable for detecting other biomarkers, thereby extending its applicability to the diagnosis of various cancer types.