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Single Cell Analysis Reveals the Presence of Novel Intermediate Cells in Both Mice and Patients With Severe MASLD
Marica Meroni1, Paride Pelucchi2, Erika Paolini1
1Medicine and Metabolic Diseases, Fondazione IRCCS Cà Granda, Ospedale Maggiore Policlinico, Milan, Italy.
Abstract:
The landscape of cellular abnormalities occurring during metabolic dysfunction-associated steatotic liver disease (MASLD) is not completely clarified. We investigated cell heterogeneity involved in progressive MASLD at single cell resolution in mice fed AMLN. Single cell RNA sequencing (Sc-RNAseq) and spatial proteomics were applied to decipher cell populations and genes/pathways guiding MASLD progression. We identified 32 clusters, including hepatocytes (HEPs), hepatic stellate cells (HSCs), endothelial cells (ENDOs), Kupffer cells (KCs), and immune cells. HEPs clusters changed across disease severity, acquiring a periportal localization in MASH-fibrosis. The latter conditions were featured by ENDO clusters with higher expression of extracellular matrix (ECM) molecules, resident and recruited KCs/immune clusters with M1 polarization and HSCs with a myofibroblast phenotype. Finally, we highlight 5 hybrid populations named HSCs/ENDOs, HEPs/ENDOs, KCs/ENDOs, and HEPs/KCs which were confirmed by spatial proteomics across disease stages in mice and MASLD patients. In sum, we observed an evolution of cellular heterogeneity during MASLD and identified novel intermediate populations that feature advanced disease.
Insights
Investigating metabolic dysfunction-associated steatotic liver disease (MASLD) at single-cell resolution revealed evolving cellular heterogeneity. Novel intermediate cell populations, including hybrid types, were identified, offering new insights into advanced MASLD progression.
Area of Science:
- Hepatology
- Cell Biology
- Molecular Biology
Background:
- The cellular landscape of metabolic dysfunction-associated steatotic liver disease (MASLD) remains incompletely understood.
- Understanding cellular abnormalities is crucial for deciphering disease progression.
Purpose of the Study:
- To investigate cellular heterogeneity in progressive MASLD using single-cell resolution.
- To identify cell populations and molecular pathways driving MASLD progression.
Main Methods:
- Employed single-cell RNA sequencing (Sc-RNAseq) and spatial proteomics in mice fed AMLN.
- Analyzed cell heterogeneity across different stages of MASLD.
Main Results:
- Identified 32 distinct cell clusters, including hepatocytes (HEPs), hepatic stellate cells (HSCs), endothelial cells (ENDOs), and Kupffer cells (KCs).
- Observed dynamic changes in HEPs and identified specific phenotypes in ENDOs, KCs, and HSCs during advanced disease stages (MASH-fibrosis).
- Discovered 5 novel hybrid cell populations (e.g., HSCs/ENDOs, HEPs/ENDOs) confirmed by spatial proteomics in mice and MASLD patients.
Conclusions:
- MASLD progression is characterized by evolving cellular heterogeneity.
- Novel intermediate and hybrid cell populations are associated with advanced MASLD and may serve as biomarkers or therapeutic targets.
