Engineering iPSC-derived neutrophils to fight infection

Abstract

Abstract Neutrophils are the initial responders to infection and injury and are critical for host survival. Immunocompromised patients are prone to recurrent bacterial and fungal infections which may require granulocyte transfusion. However, these transfusions have minimal efficacy as donor material is limited and highly heterogeneous. Induced pluripotent stem cells (iPSCs) are a promising option for generating a defined therapeutic that can improve patient outcomes. Therefore, we aimed to engineer iPSC-derived neutrophils to improve recruitment and antimicrobial function. Prior research in the field has shown that iPSC-derived neutrophils have limited function compared to primary human neutrophils, likely due to inhibitory signaling pathways. To generate iPSC-derived neutrophils with enhanced capacity, we utilized CRISPR/Cas9 to delete the protein tyrosine phosphatase 1B (PTP1B), a negative regulator of neutrophil function. PTP1B−/− neutrophils displayed increased cellular migration and inflammatory cytokine signaling in response to bacterial stimuli versus WT cells. Additionally, upon selection of cells with mature neutrophil markers, these PTP1B−/− neutrophils show similar fungal killing ability and improved phagocytosis compared to WT. Thus, these data indicate that deletion of PTP1B can improve the antimicrobial function of iPSC neutrophils and highlights the value of our system for genetically engineering iPSC-derived neutrophils. Supported by grants from the NIH (5R01AI134749-03 and Microbes in Health and Disease T32 NIAID: T32AI055397)