Peiyu Qian,1,2,* Jie Sun,3,* Zhenya Zhao,4 Peihua Lu5 

1Institutes of Biomedical Sciences, Fudan University, Shanghai, 200032, People’s Republic of China; 2Minhang Fudan Medical Education Research Center, Minhang District Central Hospital, Fudan University, Shanghai, 201100, People’s Republic of China; 3Center of Clinical Research, the Affiliated Wuxi People’s Hospital of Nanjing Medical University, Wuxi Medical Center, Nanjing Medical University, Wuxi, 214023, People’s Republic of China; 4International Medical Care Center, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 201620, People’s Republic of China; 5Department of Oncology, the Affiliated Wuxi People’s Hospital of Nanjing Medical University, Wuxi Medical Center, Nanjing Medical University, Wuxi, 214023, People’s Republic of China

*These authors contributed equally to this work

Correspondence: Peihua Lu, Department of Oncology, The Affiliated Wuxi People’s Hospital of Nanjing Medical University, Wuxi Medical Center, Nanjing Medical University, Wuxi, 214023, People’s Republic of China, Tel +8613918164127, Email lphty1_1@163.com Zhenya Zhao, International medical care Center, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 201620, People’s Republic of China, Tel +8615221091968, Email 15221091968@163.com

Objective: Gastric cancer represents a significant global health challenge due to its high prevalence and mortality rates, largely attributed to the limitations of current screening methods, such as endoscopy, which impede early diagnosis. This study presents an innovative method for early detection by identifying exfoliated tumor cells in gastric lavage, aiming to overcome challenges related to patient compliance and the variability in endoscopist expertise.
Methods: Hexokinase 2 (HK2), a metabolic marker, was utilized to identify exfoliated tumor cells with heightened glycolytic activity in gastric lavage fluid. The malignancy of these HK2-positive, high-glycolytic tumor cells was further validated using single-cell sequencing (SCS), specifically through genome-wide copy number variation analysis.
Results: A total of 60 individuals were assessed, including 10 patients with gastric cancer (9 at stage IA and 1 at stage IIA), 26 patients with precancerous lesions, 15 patients with benign gastric conditions, and 9 healthy controls. The HK2 assay demonstrated an 80% diagnostic sensitivity for stages IA and IIA of gastric cancer and a 96% diagnostic specificity in distinguishing benign conditions from healthy controls. Importantly, the assay exhibited 57% sensitivity for cases of severe dysplasia, underscoring its potential for early gastric cancer detection and preventive diagnostics.
Conclusion: The study highlights the feasibility of a novel gastric lavage-based HK2 assay, complemented by SCS for malignancy confirmation, as a highly accurate method for the early detection of gastric cancer. This approach offers a promising alternative to traditional gastroscopy, particularly for early-stage disease, potentially enhancing detection rates and improving patient outcomes.

Keywords: copy number variation, early diagnosis, gastric cancer, single cell sequencing, Warburg effect