Deciphering intratumoral heterogeneity of hepatocellular carcinoma with microvascular invasion with radiogenomic analysis

Abstract

Background and aims: The recurrence and metastasis of hepatocellular carcinoma (HCC) are mainly caused by microvascular invasion (MVI). Our study aimed to uncover the cellular atlas of MVI⁺ HCC and investigate the underlying immune infiltration patterns with radiomics features.

Methods: Three MVI positive HCC and three MVI negative HCC samples were collected for single-cell RNA-seq analysis. 26 MVI positive HCC and 30 MVI negative HCC tissues were underwent bulk RNA-seq analysis. For radiomics analysis, radiomics features score (Radscore) were built using preoperative contrast MRI for MVI prediction and overall survival prediction. We deciphered the metabolism profiles of MVI⁺ HCC using scMetabolism and scFEA. The correlation of Radscore with the level of APOE⁺ macrophages and iCAFs was identified. Whole Exome Sequencing (WES) was applied to distinguish intrahepatic metastasis (IM) and multicentric occurrence (MO). Transcriptome profiles were compared between IM and MO.

Results: Elevated levels of APOE+ macrophages and iCAFs were detected in MVI⁺ HCC. There was a strong correlation between the infiltration of APOE⁺ macrophages and iCAFs, as confirmed by immunofluorescent staining. MVI positive tumors exhibited increased lipid metabolism, which was attributed to the increased presence of APOE+ macrophages. APOE⁺ macrophages and iCAFs were also found in high levels in IM, as opposed to MO. The difference of infiltration level and Radscore between two nodules in IM was relatively small. Furthermore, we developed Radscore for predicting MVI and HCC prognostication that were also able to predict the level of infiltration of APOE⁺ macrophages and iCAFs.

Conclusion: This study demonstrated the interactions of cell subpopulations and distinct metabolism profiles in MVI⁺ HCC. Besides, MVI prediction Radscore and MVI prognostic Radscore were highly correlated with the infiltration of APOE⁺ macrophages and iCAFs, which helped to understand the biological significance of radiomics and optimize treatment strategy for MVI⁺ HCC.

Keywords: Hepatocellular carcinoma; Microvascular invasion; Radiomics; Single-cell RNA-seq.