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Updated: May 29, 2025
Direct Induction of Hemogenic Endothelium and Blood by Overexpression of Transcription Factors in Human Pluripotent Stem Cells
Published on: December 3, 2015
GATA1-deficient human pluripotent stem cells generate neutrophils with improved antifungal immunity that is mediated
Andrew S Wagner1, Frances M Smith1, David A Bennin2
1Department of Medical Microbiology and Immunology, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin, United States of America.
Abstract:
Neutrophils are critical for host defense against fungi. However, the short life span and lack of genetic tractability of primary human neutrophils has limited in vitro analysis of neutrophil-fungal interactions. Human induced pluripotent stem cell (iPSC)-derived neutrophils (iNeutrophils) provide a genetically tractable system to study host defense responses of human neutrophils. Here, we show that deletion of the transcription factor GATA1 from human iPSCs results in iNeutrophils with improved antifungal activity against Aspergillus fumigatus. GATA1-knockout (KO) iNeutrophils have increased maturation, antifungal pattern recognition receptor expression and have improved neutrophil effector functions compared to wild-type iNeutrophils. iNeutrophils also show a shift in their metabolism following stimulation with fungal β-glucan to the pentose phosphate pathway (PPP), similar to primary human neutrophils. Furthermore, we show that deletion of the integrin CD18 attenuates the ability of GATA1-KO iNeutrophils to kill A. fumigatus but is not necessary for the metabolic shift. Collectively, these findings support iNeutrophils as a robust system to study human neutrophil antifungal immunity and has identified specific roles for CD18 in the defense response.
Insights
Human induced pluripotent stem cell-derived neutrophils (iNeutrophils) lacking GATA1 show enhanced antifungal activity against Aspergillus fumigatus. These modified neutrophils exhibit improved maturation and effector functions, offering a new model for studying fungal immunity.
Area of Science:
- Immunology
- Stem Cell Biology
- Mycology
Background:
- Neutrophils are crucial for combating fungal infections but are challenging to study due to their short lifespan and limited genetic manipulation.
- Human induced pluripotent stem cell-derived neutrophils (iNeutrophils) offer a promising, genetically tractable alternative for studying neutrophil function.
- Understanding neutrophil-fungal interactions is vital for developing effective antifungal therapies.
Purpose of the Study:
- To investigate the role of the transcription factor GATA1 in human neutrophil antifungal immunity using iPSCs.
- To characterize the antifungal activity and metabolic responses of GATA1-knockout (KO) iNeutrophils.
- To elucidate the specific functions of CD18 in GATA1-KO iNeutrophil-mediated defense against Aspergillus fumigatus.
Main Methods:
- Generation of human induced pluripotent stem cells (iPSCs) with GATA1 deletion.
- Differentiation of iPSCs into iNeutrophils and comparison with wild-type iNeutrophils.
- Assessment of iNeutrophil maturation, receptor expression, effector functions, and metabolic pathways (e.g., pentose phosphate pathway) upon fungal stimulation.
- Functional assays evaluating the killing of Aspergillus fumigatus by GATA1-KO iNeutrophils, including CD18 deletion studies.
Main Results:
- GATA1-KO iNeutrophils demonstrated enhanced antifungal activity against Aspergillus fumigatus compared to wild-type iNeutrophils.
- GATA1-KO iNeutrophils exhibited increased maturation, higher expression of antifungal pattern recognition receptors, and improved effector functions.
- iNeutrophils stimulated with fungal β-glucan shifted metabolism to the pentose phosphate pathway (PPP), mirroring primary human neutrophils.
- CD18 deletion impaired the killing of Aspergillus fumigatus by GATA1-KO iNeutrophils but did not affect the metabolic shift to the PPP.
Conclusions:
- GATA1-knockout iNeutrophils represent a valuable and genetically tractable model for studying human neutrophil antifungal immunity.
- GATA1 deficiency enhances neutrophil maturation and effector functions, leading to improved antifungal capabilities.
- CD18 plays a role in the direct killing of Aspergillus fumigatus by iNeutrophils, while the metabolic response to fungal components is independent of CD18.
