Detection of rare malignant cells in gastric lavage using Hexokinase 2 and single-cell sequencing for early-stage gastric cancer diagnosis

Abstract

Objective:Gastric cancer is a highly prevalent cancer. Endoscopy is the best way to diagnose gastric cancer at an early stage, but it relies on patient compliance and endoscopy physicians’ experience, which makes it difficult to be used as a screening method for a large population. The aim of this study is to develop a novel method for early gastric cancer diagnosis by detecting exfoliated tumor cells in gastric lavage. Methods: In our experiment, Hexokinase 2 (HK2) was firstly used as a metabolic function-associated marker to detect gastric exfoliated tumor cells engaging increased glycolysis in gastric lavage. And further the malignancy of HK2-derived high glycolytic tumor cells (hgTCs) was examinedby single-cell sequencing (SCS) by surveying genome-wide copy number variation (CNV). Results: In a study of 60 individuals including 10 gastric cancer patients (9 IA and 1 IIA), 26 precancerous lesions patients, 15 patients with benign gastric diseases, and 9 healthy controls, the HK2 test showed diagnostic sensitivity and diagnostic specificity were 80% (8/10 patients with gastric cancer IA and IIA) and 96% (23/24 patients with benign gastric diseases and healthy controls), respectively. One point that is worth paying attention to is that the diagnostic sensitivity in patients with severe dysplasia was 57% (4/7), which showed promising application prospects in gastric cancer early diagnosis and prevention. Conclusions: Thus, our results demonstrated a new approach using a gastric lavage-based HK2 assay combined with SCS validation. It has the great potential to be used for early gastric cancer detection with high accuracy, especially to improve the quality of gastroscopy at the early stage.