Role of macrophage migration inhibitory factor induced netosis in regulating CDI severity

The National Institute of Allergy and Infectious Diseases awarded the University of Cincinnati's Sponsored Research Services Division $100,224 on December 10, 2026, under the Allergy and Infectious Diseases Research program (CFDA 93.855) to investigate the role of macrophage migration inhibitory factor (MIF)-induced neutrophil extracellular trap formation (netosis) in Clostridioides difficile infection (CDI) severity and identify potential host-derived therapeutic targets.

The research addresses the problem that CDI patients treated with antibiotics face high recurrence risk, necessitating novel host-mediated therapies beyond current antibiotic-primary treatment strategies. The project examines whether neutrophil-derived extracellular traps (NETs) cause collateral intestinal epithelial cell damage during CDI and whether MIF drives netosis through an NADPH oxidase-dependent pathway. Aim 1 will determine whether NETs directly promote intestinal epithelial cell apoptosis and disruption of tight junction proteins during CDI. Aim 2 will investigate MIF's role as an upstream driver of netosis via NADPH oxidase-dependent mechanisms. By elucidating the MIF-NET axis in CDI pathogenesis, the research will establish whether this mechanism represents a viable therapeutic target.

The award is classified as a Project Grant. Performance occurs at the University of Cincinnati in Cincinnati, Ohio, with an ultimate completion date of July 31, 2028.

Abstract

Clostridioides difficile infection (CDI) is one of the most common healthcare-associated infections in the united states. The current primary treatment strategy for this infection is antibiotics. However, patients who receive antibiotics are at risk of recurrent CDI, highlighting the need for novel host-mediated therapies. Our lab has shown that neutrophils worsen c. Difficile toxin-induced intestinal epithelial cell (IEC) damage, and transcriptomic data show that genes associated with neutrophil extracellular trap formation (netosis) are upregulated in blood neutrophils after CDI. Furthermore, our novel preliminary observation shows that the presence of c. Difficile bacteria induces net formation in vitro. Additionally, we have previously demonstrated that macrophage migration inhibitory factor (MIF), a pro-inflammatory cytokine, plays a detrimental role in CDI pathogenesis. However, the underlying mechanisms by which MIF mediates CDI severity remain unknown. Published studies have shown that MIF induces netosis. Aberrant nets cause tissue damage in various pathologies. Therefore, in this project, we aim to determine whether netosis leads to collateral tissue damage in CDI and examine MIF's role as an upstream driver of this response. To investigate this objective, in aim 1, we will determine whether nets directly contribute to IEC damage during CDI by promoting apoptosis and disruption of tight junction proteins. Aim 2 will then investigate whether MIF induces netosis via an nadph oxidase- dependent pathway during CDI. By elucidating the role of MIF-mediated netosis in CDI, we aim to determine whether the MIF-nets axis represents a viable host-derived therapeutic target for CDI.