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Updated: May 10, 2025
Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry
Published on: June 26, 2018
Single-cell landscape of peripheral immune cells in MASLD/MASH
Agnes Anna Steixner-Kumar1, Diana Santacruz1, Tobias Geiger2
1Department of Global Computational Biology and Digital Sciences, Boehringer Ingelheim Pharma GmbH & Co.KG, Biberach, Germany.
Background:
Metabolic dysfunction-associated steatotic liver disease (MASLD) progresses to metabolic dysfunction-associated steatohepatitis (MASH) and is a major cause of liver cirrhosis. Although liver inflammation is the hallmark feature of MASH versus MASLD, the involvement of the peripheral immune cell compartments in disease progression is poorly understood, and single-cell profiles of peripheral immune cells in MASLD/MASH are not known.
Methods:
Patients with MASLD/MASH and healthy volunteers have been prospectively enrolled in a cross-sectional study. Patients have been histologically stratified and further characterized by liver bulk RNA sequencing (RNA-Seq). Peripheral immune cells from patients and control blood samples have been comprehensively profiled using bulk and single RNA-Seq.
Results:
Twenty-two patients with fibrosis stage less than F3 have been histologically stratified into patients with low, medium, and high disease activity scores (NAFLD activity score [NAS]). In contrast to fibrosis, the NAS group correlated with noninvasive imaging readouts and blood biomarkers of liver damage and inflammation (ALT, AST). The prevalence of type 2 diabetes and obesity increased with the NAS stage. Bulk RNA-seq profiling of patient liver biopsies revealed gene signatures that were positively and negatively associated with NAS. Known marker genes for liver fibrosis where upregulated on RNA level. Blood bulk RNA-seq showed only moderate differences in patients versus healthy controls. In contrast, single-cell analysis of white blood cells revealed multiple alterations of immune (sub-)populations, including an increased abundance of immature B cells and myeloid suppressor cells in patients with MASLD/MASH as compared to healthy controls.
Conclusions:
The study gives new insights into the pathophysiology of MASLD/MASH already manifesting relatively early in peripheral immune cell compartments. This opens new avenues for the development of new biomarker diagnostics and disease therapies.
Insights
Peripheral immune cells show early changes in metabolic dysfunction-associated steatohepatitis (MASH). This study reveals altered immune cell populations in MASH patients, offering new diagnostic and therapeutic targets for liver disease.
Area of Science:
- Immunology
- Hepatology
- Genomics
Background:
- Metabolic dysfunction-associated steatotic liver disease (MASLD) can progress to metabolic dysfunction-associated steatohepatitis (MASH), a leading cause of liver cirrhosis.
- While liver inflammation is key in MASH, peripheral immune cell roles and single-cell profiles in MASLD/MASH are not well understood.
Purpose of the Study:
- To investigate the involvement of peripheral immune cell compartments in the progression of MASLD/MASH.
- To characterize single-cell profiles of peripheral immune cells in patients with MASLD/MASH.
Main Methods:
- Prospective cross-sectional study of patients with MASLD/MASH and healthy volunteers.
- Histological stratification and liver bulk RNA sequencing (RNA-Seq) of patient biopsies.
- Comprehensive bulk and single-cell RNA-Seq profiling of peripheral immune cells from blood samples.
Main Results:
- Disease activity scores (NAS) correlated with liver damage biomarkers and increased prevalence of diabetes and obesity.
- Liver bulk RNA-seq identified gene signatures associated with NAS, including upregulated fibrosis markers.
- Single-cell analysis revealed altered immune cell populations in MASH patients, notably increased immature B cells and myeloid suppressor cells compared to controls.
Conclusions:
- MASLD/MASH pathophysiology involves peripheral immune cell alterations early in the disease process.
- These findings suggest potential new avenues for developing diagnostic biomarkers and therapeutic strategies for MASH.
