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ResearchIn-Press PreviewHepatologyImmunologyVascular biology Open Access | 10.1172/JCI193758

Vessels encapsulating tumor clusters promote non-invasive metastasis of hepatocellular carcinoma by shaping an immunosuppressive microenvironment

Bi-Yu Huang,1 Zheng-Qi Mi,1 Xiao-Yu Zhang,1 Yu-Chen Ji,1 Meng-Zhi Wu,1 Zi-Feng Cheng,1 Chen Xie,1 Shuai He,2 Jing Zhu,1 Jian-Hong Fang,1 Chong Wu,1 Bin-Kui Li,3 Yun-Fei YUAN,3 Limin Zheng,1 and Shi-Mei Zhuang1

1MOE Key Laboratory of Gene Function and Regulation, Guangdong Province Key Laboratory of Pharmaceutical Functional Genes, School of Life Sciences, State Key Laboratory of Oncology in South China, Sun Yat-sen University, Guangzhou, China

2State Key Laboratory of Oncology in South China, Guangdong Provincial Clinical Research Center for Cancer, Sun Yat-sen University Cancer Center, Guangzhou, China

3Department of Hepatobiliary Oncology, Sun Yat-sen University Cancer Center, Guangzhou, China

Find articles by Huang, B. in: PubMed | Google Scholar

1MOE Key Laboratory of Gene Function and Regulation, Guangdong Province Key Laboratory of Pharmaceutical Functional Genes, School of Life Sciences, State Key Laboratory of Oncology in South China, Sun Yat-sen University, Guangzhou, China

2State Key Laboratory of Oncology in South China, Guangdong Provincial Clinical Research Center for Cancer, Sun Yat-sen University Cancer Center, Guangzhou, China

3Department of Hepatobiliary Oncology, Sun Yat-sen University Cancer Center, Guangzhou, China

Find articles by Mi, Z. in: PubMed | Google Scholar

1MOE Key Laboratory of Gene Function and Regulation, Guangdong Province Key Laboratory of Pharmaceutical Functional Genes, School of Life Sciences, State Key Laboratory of Oncology in South China, Sun Yat-sen University, Guangzhou, China

2State Key Laboratory of Oncology in South China, Guangdong Provincial Clinical Research Center for Cancer, Sun Yat-sen University Cancer Center, Guangzhou, China

3Department of Hepatobiliary Oncology, Sun Yat-sen University Cancer Center, Guangzhou, China

Find articles by Zhang, X. in: PubMed | Google Scholar

1MOE Key Laboratory of Gene Function and Regulation, Guangdong Province Key Laboratory of Pharmaceutical Functional Genes, School of Life Sciences, State Key Laboratory of Oncology in South China, Sun Yat-sen University, Guangzhou, China

2State Key Laboratory of Oncology in South China, Guangdong Provincial Clinical Research Center for Cancer, Sun Yat-sen University Cancer Center, Guangzhou, China

3Department of Hepatobiliary Oncology, Sun Yat-sen University Cancer Center, Guangzhou, China

Find articles by Ji, Y. in: PubMed | Google Scholar

1MOE Key Laboratory of Gene Function and Regulation, Guangdong Province Key Laboratory of Pharmaceutical Functional Genes, School of Life Sciences, State Key Laboratory of Oncology in South China, Sun Yat-sen University, Guangzhou, China

2State Key Laboratory of Oncology in South China, Guangdong Provincial Clinical Research Center for Cancer, Sun Yat-sen University Cancer Center, Guangzhou, China

3Department of Hepatobiliary Oncology, Sun Yat-sen University Cancer Center, Guangzhou, China

Find articles by Wu, M. in: PubMed | Google Scholar

1MOE Key Laboratory of Gene Function and Regulation, Guangdong Province Key Laboratory of Pharmaceutical Functional Genes, School of Life Sciences, State Key Laboratory of Oncology in South China, Sun Yat-sen University, Guangzhou, China

2State Key Laboratory of Oncology in South China, Guangdong Provincial Clinical Research Center for Cancer, Sun Yat-sen University Cancer Center, Guangzhou, China

3Department of Hepatobiliary Oncology, Sun Yat-sen University Cancer Center, Guangzhou, China

Find articles by Cheng, Z. in: PubMed | Google Scholar

1MOE Key Laboratory of Gene Function and Regulation, Guangdong Province Key Laboratory of Pharmaceutical Functional Genes, School of Life Sciences, State Key Laboratory of Oncology in South China, Sun Yat-sen University, Guangzhou, China

2State Key Laboratory of Oncology in South China, Guangdong Provincial Clinical Research Center for Cancer, Sun Yat-sen University Cancer Center, Guangzhou, China

3Department of Hepatobiliary Oncology, Sun Yat-sen University Cancer Center, Guangzhou, China

Find articles by Xie, C. in: PubMed | Google Scholar

1MOE Key Laboratory of Gene Function and Regulation, Guangdong Province Key Laboratory of Pharmaceutical Functional Genes, School of Life Sciences, State Key Laboratory of Oncology in South China, Sun Yat-sen University, Guangzhou, China

2State Key Laboratory of Oncology in South China, Guangdong Provincial Clinical Research Center for Cancer, Sun Yat-sen University Cancer Center, Guangzhou, China

3Department of Hepatobiliary Oncology, Sun Yat-sen University Cancer Center, Guangzhou, China

Find articles by He, S. in: PubMed | Google Scholar |

1MOE Key Laboratory of Gene Function and Regulation, Guangdong Province Key Laboratory of Pharmaceutical Functional Genes, School of Life Sciences, State Key Laboratory of Oncology in South China, Sun Yat-sen University, Guangzhou, China

2State Key Laboratory of Oncology in South China, Guangdong Provincial Clinical Research Center for Cancer, Sun Yat-sen University Cancer Center, Guangzhou, China

3Department of Hepatobiliary Oncology, Sun Yat-sen University Cancer Center, Guangzhou, China

Find articles by Zhu, J. in: PubMed | Google Scholar

1MOE Key Laboratory of Gene Function and Regulation, Guangdong Province Key Laboratory of Pharmaceutical Functional Genes, School of Life Sciences, State Key Laboratory of Oncology in South China, Sun Yat-sen University, Guangzhou, China

2State Key Laboratory of Oncology in South China, Guangdong Provincial Clinical Research Center for Cancer, Sun Yat-sen University Cancer Center, Guangzhou, China

3Department of Hepatobiliary Oncology, Sun Yat-sen University Cancer Center, Guangzhou, China

Find articles by Fang, J. in: PubMed | Google Scholar |

1MOE Key Laboratory of Gene Function and Regulation, Guangdong Province Key Laboratory of Pharmaceutical Functional Genes, School of Life Sciences, State Key Laboratory of Oncology in South China, Sun Yat-sen University, Guangzhou, China

2State Key Laboratory of Oncology in South China, Guangdong Provincial Clinical Research Center for Cancer, Sun Yat-sen University Cancer Center, Guangzhou, China

3Department of Hepatobiliary Oncology, Sun Yat-sen University Cancer Center, Guangzhou, China

Find articles by Wu, C. in: PubMed | Google Scholar |

1MOE Key Laboratory of Gene Function and Regulation, Guangdong Province Key Laboratory of Pharmaceutical Functional Genes, School of Life Sciences, State Key Laboratory of Oncology in South China, Sun Yat-sen University, Guangzhou, China

2State Key Laboratory of Oncology in South China, Guangdong Provincial Clinical Research Center for Cancer, Sun Yat-sen University Cancer Center, Guangzhou, China

3Department of Hepatobiliary Oncology, Sun Yat-sen University Cancer Center, Guangzhou, China

Find articles by Li, B. in: PubMed | Google Scholar

1MOE Key Laboratory of Gene Function and Regulation, Guangdong Province Key Laboratory of Pharmaceutical Functional Genes, School of Life Sciences, State Key Laboratory of Oncology in South China, Sun Yat-sen University, Guangzhou, China

2State Key Laboratory of Oncology in South China, Guangdong Provincial Clinical Research Center for Cancer, Sun Yat-sen University Cancer Center, Guangzhou, China

3Department of Hepatobiliary Oncology, Sun Yat-sen University Cancer Center, Guangzhou, China

Find articles by YUAN, Y. in: PubMed | Google Scholar

1MOE Key Laboratory of Gene Function and Regulation, Guangdong Province Key Laboratory of Pharmaceutical Functional Genes, School of Life Sciences, State Key Laboratory of Oncology in South China, Sun Yat-sen University, Guangzhou, China

2State Key Laboratory of Oncology in South China, Guangdong Provincial Clinical Research Center for Cancer, Sun Yat-sen University Cancer Center, Guangzhou, China

3Department of Hepatobiliary Oncology, Sun Yat-sen University Cancer Center, Guangzhou, China

Find articles by Zheng, L. in: PubMed | Google Scholar |

1MOE Key Laboratory of Gene Function and Regulation, Guangdong Province Key Laboratory of Pharmaceutical Functional Genes, School of Life Sciences, State Key Laboratory of Oncology in South China, Sun Yat-sen University, Guangzhou, China

2State Key Laboratory of Oncology in South China, Guangdong Provincial Clinical Research Center for Cancer, Sun Yat-sen University Cancer Center, Guangzhou, China

3Department of Hepatobiliary Oncology, Sun Yat-sen University Cancer Center, Guangzhou, China

Find articles by Zhuang, S. in: PubMed | Google Scholar |

Published January 6, 2026 - More info

J Clin Invest. https://doi.org/10.1172/JCI193758.
Copyright © 2026, Huang et al. This work is licensed under the Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
Published January 6, 2026 - Version history
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Abstract

Vessels encapsulating tumor clusters (VETC), a distinct vascular pattern in hepatocellular carcinoma (HCC), facilitates non-invasive metastasis in whole cluster. The interaction between VETC and tumor microenvironment requires exploration. Here, we found that compared to human Non-VETC-HCCs, VETC-tumors exhibited more PD1+CD8+ T cells and Tregs, especially TNFRSF4+Tregs and Ki67+Tregs which showed increased immunosuppressive and proliferative activity. Such immunosuppressive status was also detected in tumor emboli of VETC-HCCs, and Treg density in emboli was positively associated with metastatic cell proliferation. VETC-HCCs revealed abundance correlation, closer spatial proximity, and stronger immunosuppressive ligand-receptor interactions between TNFRSF4+Tregs/Ki67+Tregs and PD1+CD8+ T cells. Depleting Tregs in mice reduced PD1+CD8+ T cells in primary lesions, tumor emboli and metastatic foci of VETC-allografts, and attenuated allograft metastasis. TGF-β1 levels were upregulated in endothelial cells of VETC-HCCs and associated with TNFRSF4+Tregs/Ki67+Tregs enrichment. Disrupting VETC formation decreased endothelial TGF-β1 expression, and reduced TNFRSF4+Tregs/Ki67+Tregs, PD1+CD8+ T cells, Treg/CD8+ T cells ratio. Collectively, VETC may enhance Tregs’ activity via TGF-β1, while Tregs promote and sustain CD8+ T cell exhaustion through immune inhibitory ligand-receptor interaction, thereby shaping immunosuppressive microenvironment and enabling tumor cluster to carry such niche to disseminate. These findings disclose mechanisms of tumor immune microenvironment formation and provide rationales for precision medicine.

Graphical Abstract
graphical abstract
Supplemental material

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View Supplemental Table 1. The clinical characteristics of 12 HCC cases used for single-cell sequencing. Related to Supplemental Figure 1A-C.

View Supplemental Table 2. Comparing the clinical characteristics of Non-VETC and VETC HCCs used for single-cell sequencing. Related to Supplemental Figure 1A-C.

View Supplemental Table 3. Marker genes of 8 major cell clusters. Related to Figure 1A.

View Supplemental Table 4. Marker genes of T and NK cell clusters. Related to Figure 1B.

View Supplemental Table 5. Tumor-specific Treg genes. Related to Figure 2, A and G.

View Supplemental Table 6. Marker genes of Treg clusters. Related to Figure 2,C and F, Supplemental Figure 6B.

View Supplemental Table 7. T cell proliferation signature gene for Treg_MKI67 cluster. Related to Supplemental Figure 6D.

View Supplemental Table 8. Differentially expressed genes between cytotoxic and exhausted CD8+ T cells used for pseudotime trajectory analysis. Related to Supplemental Figure 7A.

View Supplemental Table 9. Cytotoxic and exhausted CD8+ T cell gene set. Related to Figure 3B, Supplemental Figure 7B and 7D.

View Supplemental Table 10. Immunoregulatory genes analyzed in tumor endothelial cells. Related to Figure 6E.

View Supplemental Table 11. The clinical characteristics in Non-VETC and VETC groups used for immunohistochemical staining. Related to Figure 1-4 and Figure 6.

View Supplemental Table 12. Antibodies used in studies.

Version history
  • Version 1 (January 6, 2026): In-Press Preview
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