Characterization of Gut Microbiota and Exploration of Potential Predictive Model for Hepatocellular Carcinoma Microvascular Invasion

Ningning Zhang; Zeyu Wang; Jiayu Lv; Shuwen Zhang; Yang Liu; Tian Liu; Wang Li; Lan Gong; Xiaodong Zhang; Emad M El-Omar; Wei Lu

doi:10.3389/fmed.2022.836369

Characterization of Gut Microbiota and Exploration of Potential Predictive Model for Hepatocellular Carcinoma Microvascular Invasion

Front Med (Lausanne). 2022 Mar 17:9:836369. doi: 10.3389/fmed.2022.836369. eCollection 2022.

Authors

Ningning Zhang^{1

2}, Zeyu Wang^{1

2}, Jiayu Lv³, Shuwen Zhang^{1

2}, Yang Liu^{1

2}, Tian Liu^{1

2}, Wang Li^{1

2}, Lan Gong^{4

5}, Xiaodong Zhang⁶, Emad M El-Omar^{4

5}, Wei Lu^{1

2}

Affiliations

¹ Department of Hepatobiliary Oncology, Liver Cancer Center, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin's Clinical Research Center for Cancer, Tianjin Medical University, Tianjin, China.
² Tianjin's Clinical Research Center for Cancer, Tianjin Medical University, Tianjin, China.
³ Department of Hepatology, Tianjin Third Central Hospital, Tianjin, China.
⁴ Department of Medicine, University of New South Wales, Sydney, NSW, Australia.
⁵ St George & Sutherland Clinical School, Microbiome Research Centre, University of New South Wales, Sydney, NSW, Australia.
⁶ Key Laboratory of Cancer Prevention and Therapy, Department of Gastrointestinal Cancer Biology, Liver Cancer Center, National Clinical Research Center for Cancer, Tianjin Medical University Cancer Institute and Hospital, Tianjin Medical University, Tianjin, China.

Abstract

Background: The association between gut microbiota and microvascular invasion (MVI) in patients with hepatocellular carcinoma (HCC) remains unclarified. Hence, the microbiome analysis of patients with HCC might predict MVI development as an accurate, non-invasive, and convenient assessment. The aim of this study was to investigate the characteristics of gut microbiota in patients with HCC-MVI and establish a microbial prediction model of HCC-MVI based on a microbiome study.

Methods: Fecal samples were collected from 59 patients with HCC (24 of the total with MVI disease and 16 healthy controls) and were further analyzed by 16S rRNA amplicon sequencing followed by a comprehensive bioinformatic analysis. The diagnostic performance of microbiome characteristics in predicting MVI was assessed by receiver operating characteristic (ROC) curves. The correlation between gut microbiota and tumor microenvironment (TME) in the HCC-MVI group was further analyzed by using immunohistochemistry and immunofluorescence assay.

Results: A significant differentiation trend of microbiota composition and structure was observed between the HCC-MVI group and those without vascular invasion (HCC-NVI). Compared with HCC-NVI group and healthy controls, gut bacteria Klebsiella, Proteobacteria, Prevotellaceae, and Enterobacteriaceae were significantly enriched, whereas Firmicutes, Ruminococcus, and Monoglobaceae were significantly decreased in patients with HCC-MVI. Klebsiella was considered to be the key microbiome signature for patients with HCC-MVI. The area under the curve (AUC) of the established HCC-MVI microbial prediction model was 94.81% (95% CI: 87.63-100%). The percentage of M2-type tumor-associated macrophages (TAMs) was increased in the HCC-MVI group compared with the HCC-NVI group (p < 0.001). M2-type TAMs in TME were negatively correlated with Shannon and Simpson index of HCC-MVI gut microbiota (all p < 0.01). In addition, predicted KEGG pathways showed that the functional differences in the metabolic pathways of microbiota varied among the groups.

Conclusion: The results indicated that differences existed in the fecal microbiome of patients with HCC-MVI and healthy controls. The prediction model of HCC-MVI established with certain gut bacterial signatures may have the potential to predict HCC-MVI outcome, and the characteristics of the fecal microbiome in patients with HCC may be associated with TME, though future larger-cohort studies are required to validate this supposition.

Keywords: M2-type TAM; gut microbiota; hepatocellular carcinoma; mTOR signaling pathway; microvascular invasion; predictive model; tumor microenvironment.