Featured Intestinal Microbiota Associated With Hepatocellular Carcinoma in Various Liver Disease States

Xiu Sun¹Zhewen Zhou¹Xin Chi¹Danying Cheng¹YuanYuan ZHANG²Yifan Xu¹Yanxu Hao¹Ying Duan¹Wei Li¹Yingying Zhao¹Shunai Liu²Ming Han²Xi Wang²Song Yang¹Calvin Q Pan^3*Huichun Xing^1*

• ¹Beijing Ditan Hospital, Capital Medical University, Beijing, China
• ²Beijing Key Laboratory of Emerging Infectious Diseases, Institute of Infectious Disease, Beijing Ditan Hospital, Capital Medical University, Beijing, China
• ³Division of Gastroenterology and Hepatology, NYU Langone Health, New York University School of Medicine, New york, United States

Abstract Objective: This study aimed to identify distinct intestinal microbiota associated with hepatocellular carcinoma (HCC) and to construct a predictive model for HCC. Methods: A case-control study was conducted including patients with chronic hepatitis B (CHB), liver cirrhosis (LC), HCC, and healthy controls (HC). Fecal 16S rDNA sequences were analyzed using bioinformatics approaches. Specific intestinal microbiota were identified through stratified analysis, and a predictive model was subsequently constructed. Results: A total of 152 subjects were enrolled, including CHB (n = 33), LC (n = 59; 25 compensated cirrhosis, CC; 34 decompensated cirrhosis, DC), HCC (n = 30; 5 CHB-HCC, 9 CC-HCC, and 16 DC-HCC), and HC (n = 30). A significant overall difference in alpha diversity was observed across the groups (Chao1: P = 0.010, ε²= 0.056; ACE: P = 0.016, ε²= 0.049). In the CHB-HCC, CC-HCC, and DC-HCC groups, the abundance of Bacteroides, Prevotella, and Faecalibacterium gradually decreased, whereas Klebsiella, Haemophilus, and Streptococcus increased. Comparison of CHB vs. CHB-HCC, CC vs. CC-HCC, and DC vs. DC-HCC revealed consistent microbial shifts across disease stages. In particular, Roseburia, Veillonella, Megasphaera, and Paraprevotella were increased irrespective of liver disease stage. By combining microbiota profiles with clinical indicators, we developed a predictive nomogram that achieved an AUC of 0.865 in the training cohort and 0.848 in the external validation cohort. Conclusion: Intestinal microbiota were associated not only with liver disease stage but also with the occurrence of HCC itself. Characteristic microbiota may serve as effective biomarkers for predicting HCC.