Oberlick, Elaine, Smith, Tomas, Park, Seonmi, Bray, Mark, Antczak, Christophe, Corey, Driscoll, Sarah, Mowbray and Duncan, Nunes (2023) GATA1 deletion in human pluripotent stem cells increases differentiation yield and maturity of neutrophils. iScience, 26 (10). p. 107804. ISSN 2589-0042

Abstract

Human pluripotent stem cell (hPSC)-derived tissues can be used to model diseases in cell types that are challenging to harvest and study at-scale, such as neutrophils. Neutrophil dysregulation, specifically neutrophil extracellular trap (NET) formation, plays a critical role in the prognosis and progression of multiple diseases, including COVID-19. While hPSCs can generate limitless neutrophils (iNeutrophils) to study these processes, current differentiation protocols generate heterogeneous cultures of granulocytes and precursors. Here, we describe a method to improve iNeutrophil differentiations through the deletion of GATA1. GATA1 knockout (KO) iNeutrophils are nearly identical to primary neutrophils in form and function. Unlike wild-type iNeutrophils, GATA1 KO iNeutrophils generate NETs in response to the physiologic stimulant lipopolysaccharide, suggesting they are a more accurate model when performing NET inhibitor screens. Furthermore, through deletion of CYBB, we demonstrate that GATA1 KO iNeutrophils are a powerful tool in determining involvement of a given protein in NET formation.

Item Type:	Article
Date Deposited:	12 Dec 2023 00:45
Last Modified:	12 Dec 2023 00:45
URI:	https://oak.novartis.com/id/eprint/49597