Regulation of neutrophil heterogeneity and activation state using human induced pluripotent stem cell-derived neutrophils

About This Grant

PROJECT SUMMARY Neutrophils are critical for host defense against infection. Neutrophils can have either pro- or anti-inflammatory effects and exhibit significant heterogeneity and plasticity. There is a gap in understanding neutrophil heterogeneity and function because they are short-lived ex vivo and are not genetically tractable. Induced pluripotent stem cells (iPSC)-derived human neutrophils (iNeutrophils) offer the opportunity to genetically and developmentally program neutrophils for designed properties that may be used both to understand neutrophil biology and provide an avenue for engineering neutrophils for human disease treatment. We have engineered GMP-compatible human iNeutrophils that show improved antimicrobial function. Depleting the transcription factor GATA1 from human iPSCs results in a uniform mature population of iNeutrophils. However, the mechanisms of cell fate determination that result in neutrophil maturation and the specific role of GATA1 remains unknown. This neutrophil-biased differentiation potentially mimics the phenotype of neutrophilia seen in Down syndrome patients with a short isoform of GATA1 (GATA1s), suggesting the importance of full length GATA1 in regulating myeloid cell fates and functions in vivo. Here we propose to use single-cell multi-omics and imaging to understand how GATA1 and its isoforms modulate iNeutrophil development and function, with the long-term goal of increasing understanding of the drivers of neutrophil phenotypes to enable engineering of neutrophils with improved antimicrobial functions. We are uniquely positioned with the tools in place to make rapid progress with the proposed aims because of recent progress: 1. Our single-cell and metabolic imaging data show distinct neutrophil populations, with more maturity and less heterogeneity in GATA1 KO cells. 2. Preliminary data show epigenetic changes in the GATA1 KO that may indicate rewiring of gene networks in the absence of GATA1.