Screening and Modeling of LUAD-Associated Predictive and Prognostic Genes Based on Multi-Omics Data

LUAD is widespread worldwide with onset mainly in the low-grade stage due to early screen. However, the ambiguity surrounding small-sized nodules often leads to hesitation in deciding between surgery and surveillance. Liquid biopsy which leverages the cell-free RNA (cfRNA) shows excellent classification performance among disease groups in addition to the non-invasive feature. This research utilizes liquid biopsy to construct an effective screening system to aid with the further differentiation of benign diseases from lung cancers. Overall design: Peripheral whole blood samples were collected from individuals before therapy or surgery using EDTA-coated vacutainer tubes. Plasma was separated within 2 hr after collection by centrifuge at 1,900 g for 10 min at 4 ?. All plasma samples were aliquoted and stored at -80°C. CfRNAs were extracted using QIAzol Lysis Reagent (QIAGEN, Beijing, China), and DNA contamination was removed by Recombinant DNase I (RNase-free) (TAKARA, Beijing, China). End repair and polyadenylation were performed using E. coli Poly(A) Polymerase (NEB, Beijing, China), and T4 Polynucleotide Kinase (NEB, Beijing, China). Reverse transcription and libraries were prepared according to DETECTOR-seq protocol with minor modifications by replacing original TSO with uracil-containing TSO and extra digestion with Uracil-DNA Glycosylase (NEB, Beijing, China) after reverse transcription. Library quantification was performed by Qubit dsDNA HS Kit. Library fragment size and quality were checked using Agilent 2100 Bioanalyzer. Libraries were sequenced on MGI Tech DNBSEQ-T7 with PE150.