Project Details

IMmuno-coagulopathic PRofiling in TAMOF (IMPRINT)

Steven Ham, MD, MPH

Summary

BACKGROUND: Multiple organ dysfunction syndrome (MODS) is a leading cause of death in critically ill children, particularly those with sepsis. Thrombocytopenia is a marker of a high risk MODS phenotype, characterized by microvascular thrombosis and endothelial injury, yet clinicians lack a validated method of identifying when low platelet counts reflect a treatable thromboinflammatory process. GAP: Using high dimensional plasma proteomics, our group identified distinct immuno coagulopathic signatures in children with thrombocytopenia associated multiple organ failure (TAMOF), suggesting a biologically meaningful and potentially targetable disease mechanism. However, these findings require functional validation to determine whether they represent a reversible pathway that could guide future therapies. HYPOTHESIS: We hypothesize that endothelialized microfluidic channels exposed to TAMOF plasma will demonstrate increased thrombosis, neutrophil extracellular trap (NET) formation, and complement activation, and that these effects can be mitigated through complement inhibition. METHODS: We will culture human umbilical vein endothelial cells in BioFlux microfluidic plates and introduce healthy donor platelets and neutrophils, followed by infusion of plasma from healthy controls and children with TAMOF. High resolution confocal imaging and immunofluorescence will quantify thrombus formation, NETosis, and endothelial complement deposition, with and without eculizumab. RESULTS: Our preliminary proteomic work, using supervised clustering and machine learning, identified convergence of IL-6, TNF-α, and IFN-γ signaling with elevated complement activity in a TAMOF predominant cluster. IMPACT: This project will provide foundational mechanistic evidence linking NET mediated complement activation to thrombocytopenia and microvascular thrombosis in pediatric MODS. By validating whether a clinically recognizable phenotype corresponds to a reversible immuno-thrombotic mechanism, this work aims to inform future diagnostic and therapeutic strategies for children with TAMOF. Website Link: https://lindelllab.org/Optional/Additional Comments: Steven Ham is a T32 research fellow at the Children’s Hospital of Philadelphia.