GATA1-deficient human pluripotent stem cells generate neutrophils with improved antifungal immunity that is mediated by the integrin CD18 DOI Creative Commons
Andrew S. Wagner, Frances M. Smith, David A. Bennin

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Окт. 11, 2024

Abstract Neutrophils are critical for host defense against fungi. However, the short life span and lack of genetic tractability primary human neutrophils has limited in vitro analysis neutrophil-fungal interactions. Human induced pluripotent stem cell (iPSC)-derived (iNeutrophils) a genetically tractable alternative to neutrophils. Here, we show that deletion transcription factor GATA1 from iPSCs results iNeutrophils with improved antifungal activity Aspergillus fumigatus . knockout (KO) have increased maturation, pattern recognition receptor expression more readily execute neutrophil effector functions compared wild-type iNeutrophils. also shift their metabolism following stimulation fungal β-glucan, including an upregulation pentose phosphate pathway (PPP), similar Furthermore, integrin CD18 attenuates ability GATA1-KO kill A. but is not necessary PPP. Collectively, these findings support as robust system study immunity identified specific roles response. Author Summary important first responders infections, understanding essential better elucidating disease dynamics. Primary lived do permit manipulation, limiting use interactions analyses. In this report GATA1-deficient generate mature than display pathogen We receptors cells at levels comparable Deletion blocks control growth , demonstrating role iNeutrophil activity. model immunity.

Язык: Английский

GATA1-deficient human pluripotent stem cells generate neutrophils with improved antifungal immunity that is mediated by the integrin CD18 DOI Creative Commons
Andrew S. Wagner, Frances M. Smith, David A. Bennin

и другие.

PLoS Pathogens, Год журнала: 2025, Номер 21(2), С. e1012654 - e1012654

Опубликована: Фев. 3, 2025

Neutrophils are critical for host defense against fungi. However, the short life span and lack of genetic tractability primary human neutrophils has limited in vitro analysis neutrophil-fungal interactions. Human induced pluripotent stem cell (iPSC)-derived (iNeutrophils) provide a genetically tractable system to study responses neutrophils. Here, we show that deletion transcription factor GATA1 from iPSCs results iNeutrophils with improved antifungal activity Aspergillus fumigatus. GATA1-knockout (KO) have increased maturation, pattern recognition receptor expression neutrophil effector functions compared wild-type iNeutrophils. also shift their metabolism following stimulation fungal β-glucan pentose phosphate pathway (PPP), similar Furthermore, integrin CD18 attenuates ability GATA1-KO kill A. fumigatus but is not necessary metabolic shift. Collectively, these findings support as robust immunity identified specific roles response.

Язык: Английский

Процитировано

0

Mitochondrial metabolism is rapidly re-activated in mature neutrophils to support stimulation-induced response DOI Creative Commons
Jorgo Lika, James A. Votava, Rupsa Datta

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2025, Номер unknown

Опубликована: Фев. 8, 2025

Abstract Neutrophils are highly abundant innate immune cells that constantly produced from myeloid progenitors in the bone marrow. Differentiated neutrophils can perform an arsenal of effector functions critical for host defense. This study aims to quantitatively understand neutrophil mitochondrial metabolism throughout differentiation and activation, elucidate impact on functions. To metabolic remodeling differentiation, murine ER-Hoxb8 progenitor-derived human induced pluripotent stem cell-derived were assessed as models. upon differentiated primary activated with various stimuli, including ionomycin, MSU crystals, PMA. Characterization cellular by isotopic tracing, extracellular flux analysis, metabolomics, fluorescence-lifetime imaging microscopy revealed dynamic changes metabolism. As mature, decreases drastically, energy production is fully offloaded oxidative phosphorylation, glucose oxidation through TCA cycle substantially reduced. Nonetheless, mature retain capacity Upon stimulation certain rapidly activated. Mitochondrial pyruvate carrier inhibitors reduce this re-activation inhibit release traps. also impacts redox status, migration, apoptosis without significantly changing overall bioenergetics. Together, these results demonstrate dynamically remodeled plays a significant role function fate. Furthermore, findings point therapeutic potential range conditions where dysregulated response drives inflammation contributes pathology.

Язык: Английский

Процитировано

0

Micro blood analysis technology (μBAT): multiplexed analysis of neutrophil phenotype and function from microliter whole blood samples DOI

Terry D. Juang,

Jeremiah Riendeau,

Peter Geiger

и другие.

Lab on a Chip, Год журнала: 2024, Номер 24(17), С. 4198 - 4210

Опубликована: Янв. 1, 2024

There is an ongoing need to do more with less and provide highly multiplexed analysis from limited sample volumes. Improved "sample sparing" assays would have a broad impact across pediatric other rare type studies in addition enabling sequential sampling. This capability advance both clinical basic research applications. Here we report the micro blood technology (μBAT), microfluidic platform that supports of neutrophils single drop blood. We demonstrate orthogonal capabilities μBAT including functional (phagocytosis, neutrophil extracellular traps, optical metabolic imaging) molecular (gene expression, cytokine secretion). Importantly validate our microscale using macroscale benchmark assay. compatible lancet puncture or microdraw devices, its design facilitates rapid operations without for specialized equipment. offers new method investigating function populations restricted amounts.

Язык: Английский

Процитировано

1

GATA1-deficient human pluripotent stem cells generate neutrophils with improved antifungal immunity that is mediated by the integrin CD18 DOI Creative Commons
Andrew S. Wagner, Frances M. Smith, David A. Bennin

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Окт. 11, 2024

Abstract Neutrophils are critical for host defense against fungi. However, the short life span and lack of genetic tractability primary human neutrophils has limited in vitro analysis neutrophil-fungal interactions. Human induced pluripotent stem cell (iPSC)-derived (iNeutrophils) a genetically tractable alternative to neutrophils. Here, we show that deletion transcription factor GATA1 from iPSCs results iNeutrophils with improved antifungal activity Aspergillus fumigatus . knockout (KO) have increased maturation, pattern recognition receptor expression more readily execute neutrophil effector functions compared wild-type iNeutrophils. also shift their metabolism following stimulation fungal β-glucan, including an upregulation pentose phosphate pathway (PPP), similar Furthermore, integrin CD18 attenuates ability GATA1-KO kill A. but is not necessary PPP. Collectively, these findings support as robust system study immunity identified specific roles response. Author Summary important first responders infections, understanding essential better elucidating disease dynamics. Primary lived do permit manipulation, limiting use interactions analyses. In this report GATA1-deficient generate mature than display pathogen We receptors cells at levels comparable Deletion blocks control growth , demonstrating role iNeutrophil activity. model immunity.

Язык: Английский

Процитировано

1