Journal Article

Multi-Scale Spatial Analysis of the Tumor Microenvironment Reveals Features of Cabozantinib and Nivolumab Efficacy in Hepatocellular Carcinoma

Popel, Aleksander S., Ho, Won Jin, Mi, Haoyang, Yarchoan, Mark

2022

Overview

Concomitant inhibition of vascular endothelial growth factor (VEGF) and programmed cell death protein 1 (PD-1) or its ligand PD-L1 is a standard of care for patients with advanced hepatocellular carcinoma (HCC), but only a minority of patients respond, and responses are usually transient. Understanding the effects of therapies on the tumor microenvironment (TME) can provide insights into mechanisms of therapeutic resistance. 14 patients with HCC were treated with the combination of cabozantinib and nivolumab through the Johns Hopkins Sidney Kimmel Comprehensive Cancer Center. Among them, 12 patients (5 responders + 7 non-responders) underwent successful margin negative resection and are subjects to tissue microarray (TMA) construction containing 37 representative tumor region cores. Using the TMAs, we performed imaging mass cytometry (IMC) with a panel of 27-cell lineage and functional markers. All multiplexed images were then segmented to generate a single-cell dataset that enables (1) tumor-immune compartment analysis and (2) cell community analysis based on graph-embedding methodology. Results from these hierarchies are merged into response-associated biological process patterns. Image processing on 37 multiplexed-images discriminated 59,453 cells and was then clustered into 17 cell types. Compartment analysis showed that at immune-tumor boundaries from NR, PD-L1 level on tumor cells is significantly higher than remote regions; however, Granzyme B expression shows the opposite pattern. We also identify that the close proximity of CD8 T cells to arginase 1 (Arg1 ) macrophages, rather than CD4 T cells, is a salient feature of the TME in non-responders. Furthermore, cell community analysis extracted 8 types of cell-cell interaction networks termed cellular communities (CCs). We observed that in non-responders, macrophage-enriched CC (MCC) and lymphocyte-enriched CC (LCC) strongly communicate with tumor CC, whereas in responders, such communications were undermined by the engagement between MCC and LCC. These results demonstrate the feasibility of a novel application of multiplexed image analysis that is broadly applicable to quantitative analysis of pathology specimens in immuno-oncology and provides further evidence that CD163 Arg1 macrophages may be a therapeutic target in HCC. The results also provide critical information for the development of mechanistic quantitative systems pharmacology models aimed at predicting outcomes of clinical trials.

• Publisher: Frontiers Media SA,Frontiers Media S.A
• Subject: Anilides / Apoptosis / Arginase / B7-H1 Antigen - metabolism / biomarker / Cancer immunotherapy / Carcinoma, Hepatocellular - pathology / CD163 antigen / CD4 antigen / CD8 antigen / CD8-Positive T-Lymphocytes - metabolism / Cell death / Cell lineage / Cells / Clinical trials / computational biology / Drug dosages / Embedding / Granzyme B / Hepatocellular carcinoma / Humans / Image processing / Immunology / immunotherapy / Kinases / Liver cancer / Liver Neoplasms - pathology / Lymphocytes T / Macrophages / Nivolumab - therapeutic use / Oncology / Patients / PD-1 protein / PD-L1 protein / Pyridines / Spatial Analysis / Standard of care / systems biology / Therapeutic targets / Tumor cells / Tumor Microenvironment / tumor-immune microenvironments (TIME) / Tumors / Vascular endothelial growth factor / Vascular Endothelial Growth Factor A - metabolism