Heterogeneity in intrahepatic macrophage populations and druggable target expression in patients with steatotic liver disease-related fibrosis

Omar A Saldarriaga; Timothy G Wanninger; Esteban Arroyave; Joseph Gosnell; Santhoshi Krishnan; Morgan Oneka; Daniel Bao; Daniel E Millian; Michael L Kueht; Akshata Moghe; Jingjing Jiao; Jessica I Sanchez; Heidi Spratt; Laura Beretta; Arvind Rao; Jared K Burks; Heather L Stevenson

doi:10.1016/j.jhepr.2023.100958

Heterogeneity in intrahepatic macrophage populations and druggable target expression in patients with steatotic liver disease-related fibrosis

JHEP Rep. 2023 Nov 3;6(1):100958. doi: 10.1016/j.jhepr.2023.100958. eCollection 2024 Jan.

Authors

Omar A Saldarriaga¹, Timothy G Wanninger², Esteban Arroyave¹, Joseph Gosnell¹, Santhoshi Krishnan^{3

4}, Morgan Oneka³, Daniel Bao⁵, Daniel E Millian¹, Michael L Kueht⁶, Akshata Moghe⁷, Jingjing Jiao⁸, Jessica I Sanchez⁸, Heidi Spratt⁹, Laura Beretta⁸, Arvind Rao^{3

4

10

11

12}, Jared K Burks¹³, Heather L Stevenson¹

Affiliations

¹ Department of Pathology, University of Texas Medical Branch, Galveston, TX, USA.
² Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA.
³ Department of Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, MI, USA.
⁴ Department of Electrical and Computer Engineering, Rice University, Houston, TX, USA.
⁵ School of Medicine, University of Texas Medical Branch, Galveston, TX, USA.
⁶ Department of Surgery, University of Texas Medical Branch, Galveston, TX, USA.
⁷ Department of Internal Medicine, University of Texas Medical Branch, Galveston, TX, USA.
⁸ Department of Molecular and Cellular Oncology, University of Texas MD Anderson Cancer Center, Houston, TX, USA.
⁹ Department of Biostatistics and Data Science, University of Texas Medical Branch, Galveston, TX, USA.
¹⁰ Departmen of Radiation Oncology, University of Michigan, Ann Arbor, MI, USA.
¹¹ Department of Biostatistics, University of Michigan, Ann Arbor, MI, USA.
¹² Department of Biomedical Engineering, Rice University, Ann Arbor, MI, USA.
¹³ Department of Leukemia, University of Texas MD Anderson Cancer Center, Houston, TX, USA.

Abstract

Background & aims: Clinical trials for reducing fibrosis in steatotic liver disease (SLD) have targeted macrophages with variable results. We evaluated intrahepatic macrophages in patients with SLD to determine if activity scores or fibrosis stages influenced phenotypes and expression of druggable targets, such as CCR2 and galectin-3.

Methods: Liver biopsies from controls or patients with minimal or advanced fibrosis were subject to gene expression analysis using nCounter to determine differences in macrophage-related genes (n = 30). To investigate variability among individual patients, we compared additional biopsies by staining them with multiplex antibody panels (CD68/CD14/CD16/CD163/Mac387 or CD163/CCR2/galectin-3/Mac387) followed by spectral imaging and spatial analysis. Algorithms that utilize deep learning/artificial intelligence were applied to create cell cluster plots, phenotype profile maps, and to determine levels of protein expression (n = 34).

Results: Several genes known to be pro-fibrotic (e.g. CD206, TREM2, CD163, and ARG1) showed either no significant differences or significantly decreased with advanced fibrosis. Although marked variability in gene expression was observed in individual patients with cirrhosis, several druggable targets and their ligands (e.g. CCR2, CCR5, CCL2, CCL5, and LGALS3) were significantly increased when compared to patients with minimal fibrosis. Antibody panels identified populations that were significantly increased (e.g. Mac387+), decreased (e.g. CD14+), or enriched (e.g. interactions of Mac387) in patients that had progression of disease or advanced fibrosis. Despite heterogeneity in patients with SLD, several macrophage phenotypes and druggable targets showed a positive correlation with increasing NAFLD activity scores and fibrosis stages.

Conclusions: Patients with SLD have markedly varied macrophage- and druggable target-related gene and protein expression in their livers. Several patients had relatively high expression, while others were like controls. Overall, patients with more advanced disease had significantly higher expression of CCR2 and galectin-3 at both the gene and protein levels.

Impact and implications: Appreciating individual differences within the hepatic microenvironment of patients with SLD may be paramount to developing effective treatments. These results may explain why such a small percentage of patients have responded to macrophage-targeting therapies and provide additional support for precision medicine-guided treatment of chronic liver diseases.

Keywords: Cardiometabolic risk factors; Chemokines; Infiltrating monocytes; Inflammation; MASLD; Multispectral imaging; NanoString nCounter; Visiopharm.

Grants and funding

R01 DK125730/DK/NIDDK NIH HHS/United States