Integrated single-cell multiomics reveals neutrophil-driven immune-metabolic reprogramming of Kupffer cells via thrombospondin-1/CD36 after surgical stress.
Integrated single-cell multiomics identifies a dominant neutrophil-Kupffer cell communication axis mediated by neutrophil-derived thrombospondin-1 (TSP-1) engaging CD36 on Kupffer cells that drives immune-metabolic reprogramming and sterile liver injury following surgical stress, with pharmacologic inhibition of TSP-1 attenuating liver injury in vivo.
Key Findings
Methods
Single-cell RNA sequencing, spatial transcriptomics, high-dimensional spectral flow cytometry, and metabolomics together resolved the hepatic immune landscape following ischemia/reperfusion (I/R) injury at cellular and spatial resolution.
The study integrated four distinct omics/analytical platforms to characterize the hepatic immune microenvironment after surgical stress.
The model used was liver ischemia/reperfusion (I/R) injury, characterized by robust neutrophil infiltration and immune activation.
The integrated approach enabled identification of intercellular circuits that spatially orchestrate sterile inflammatory responses within the hepatic immune microenvironment.
Extensive immune remodeling was confirmed across the hepatic immune landscape following I/R.
Results
A dominant neutrophil-Kupffer cell communication axis mediated by neutrophil-derived thrombospondin-1 (TSP-1) selectively engaging CD36 on Kupffer cells was identified as a central driver of hepatic immune responses after I/R.
TSP-1 was identified as being derived from neutrophils and acting as the primary ligand in this intercellular signaling axis.
CD36 on Kupffer cells was identified as the selective receptor for neutrophil-derived TSP-1.
This TSP-1/CD36 axis was identified as a conserved and targetable mediator of sterile liver injury.
Cross-species integration with human liver transplant datasets demonstrated conserved upregulation of the TSP-1/CD36 axis following reperfusion.
Results
TSP-1/CD36 interaction drives coordinated immune-metabolic reprogramming in Kupffer cells, characterized by suppression of oxidative phosphorylation, enhanced glycolysis, and remodeling of sphingolipid metabolism.
The metabolic reprogramming in Kupffer cells included suppression of oxidative phosphorylation and enhanced glycolysis.
Sphingolipid metabolism was remodeled, including accumulation of hexosylceramides in Kupffer cells.
Metabolomics was used to characterize these metabolic changes.
This immune-metabolic reprogramming was described as 'coordinated,' suggesting a systematic functional shift in Kupffer cell biology driven by the neutrophil-derived signal.
Results
Spatial analyses revealed preferential neutrophil-Kupffer cell colocalization within necrotic niches of the injured liver.
Spatial transcriptomics was used to determine the anatomical organization of the neutrophil-Kupffer cell interactions.
Neutrophils and Kupffer cells were found to preferentially colocalize within necrotic niches specifically.
This spatial organization supports a model of locally organized, spatially restricted intercellular communication rather than diffuse paracrine signaling.
The findings define a 'spatially organized, neutrophil-driven immune-metabolic circuit' governing Kupffer cell reprogramming during surgical stress.
Results
Pharmacologic inhibition of TSP-1 attenuated liver injury and inflammatory responses in vivo.
TSP-1 inhibition was tested pharmacologically in an in vivo model.
The intervention resulted in attenuation of both liver injury and inflammatory responses.
This finding identifies the TSP-1/CD36 pathway as a targetable mediator of sterile liver injury.
The pharmacologic validation supports the functional importance of the neutrophil-derived TSP-1 signal in driving pathological outcomes.
Results
The TSP-1/CD36 axis was conserved across species, with upregulation confirmed in human liver transplant datasets following reperfusion.
Cross-species integration was performed between mouse I/R injury data and human liver transplant datasets.
Conserved upregulation of the TSP-1/CD36 axis was demonstrated following reperfusion in human tissue.
This cross-species conservation supports the translational relevance of findings from the murine surgical stress model.
Human liver transplantation represents a clinically significant context of hepatic I/R injury where these findings may apply.
What This Means
This research suggests that during surgical procedures involving the liver—such as liver transplantation or other surgeries requiring temporary blood flow restriction—a specific chain of cellular communication events drives dangerous inflammation and organ damage. Using multiple advanced technologies simultaneously, the researchers mapped out exactly which immune cells talk to each other and how, following a period of reduced then restored blood flow to the liver (called ischemia/reperfusion injury). They discovered that neutrophils, a type of immune cell that floods into the liver early after injury, release a protein called thrombospondin-1 (TSP-1) that directly signals to liver-resident immune cells called Kupffer cells through a receptor called CD36. This signal fundamentally changes how Kupffer cells produce energy and manage fats, shifting them toward a more inflammatory and potentially harmful state.
The study also found that these neutrophil-Kupffer cell interactions happen in specific physical locations within the liver—particularly in areas of tissue death—suggesting the damage is spatially organized rather than randomly distributed. Importantly, the same TSP-1/CD36 communication pathway was found to be active in human liver transplant patients, not just in the mouse models used in the laboratory, indicating these findings are likely relevant to human medicine. When the researchers blocked TSP-1 with a drug in mice, liver injury and inflammation were reduced, demonstrating that this pathway could potentially be targeted therapeutically.
This research suggests that the TSP-1/CD36 signaling axis between neutrophils and Kupffer cells represents a previously unappreciated but critical driver of sterile liver inflammation after surgical stress. The findings open potential new avenues for protecting the liver during high-risk surgical procedures by targeting this specific molecular pathway, which may be particularly relevant for patients undergoing liver transplantation or major liver surgery where ischemia/reperfusion injury is unavoidable.