CRISPR/Cas9-mediated loss of FERMT3 reveals kindlin-3-independent neutrophil activity in LAD-III.
7 affiliations
- 1Department of Research, Sanquin Blood Supply Foundation, Amsterdam, Netherlands.
- 2Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands.
- 3Department of Molecular and Integrative Biosciences, Faculty of Biological and Environmental Sciences, University of Helsinki, Helsinki, Finland.
- 4Institute of Molecular Oncology and Functional Genomics, School of Medicine, Technical University of Munich , Munich, Germany.
- 5Institute of Experimental Hematology School of Medicine and Health, Technical University of Munich , Munich, Germany.
- 6Center for Translational Cancer Research (TranslaTUM), School of Medicine and Health, Technical University of Munich , Munich, Germany.
- 7Department of Pediatric Immunology, Rheumatology & Infectious Diseases, Emma Children's Hospital, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands.
Abstract
Mutations in human FERMT3 (encoding kindlin-3 protein) impair β2 integrin activity in neutrophils from patients with leukocyte adhesion deficiency III (LAD-III). Yet kindlin-3's contribution to neutrophil integrin affinity regulation and function remains unclear. Using CD34+ human hematopoietic stem and progenitor cells (HSPCs) differentiated into neutrophils, we investigated how CRISPR/Cas9-mediated kindlin-3 deletion affects neutrophil characteristics. CRISPR/Cas9 targeting FERMT3 resulted in an ∼80-bp deletion within exon 2, impairing protein production. FERMT3 KO neutrophils retained normal in vitro development and phenotype with similar surface marker expression, nuclear morphology, and expansion potential compared with untreated cultured neutrophils. Loss of kindlin-3 abolished zymosan-induced NADPH oxidase activity, recapitulating the ROS production defect of LAD-III neutrophils. However, although basal adhesion was reduced, stimulus-induced integrin activity remained largely intact. FERMT3 KO neutrophils retained fMLP-driven chemotaxis while showing impaired migration in response to C5a. Together, these findings show that disruption of kindlin-3 largely recapitulates the LAD-III phenotype while exerting some effector functions upon stimulation without intact kindlin-3.
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BibTeX
@article{webbers2026crisprcas9mediated,
title = {{CRISPR/Cas9-mediated loss of FERMT3 reveals kindlin-3-independent neutrophil activity in LAD-III.}},
author = {Steven D S Webbers and Felipe Rojas-Rodriguez and Angela A F Gankema and Imran Khan and Judy Geissler and Anton T J Tool and Rupert Öllinger and Robin van Bruggen and Teunis J P van Dam and Suzanna C Fagerholm and Markus Moser and Taco W Kuijpers},
journal = {Journal of human immunity},
year = {2026},
month = oct,
doi = {10.70962/jhi.20260086},
url = {https://europepmc.org/article/MED/42905029},
note = {CC BY},
}
RIS
TY - JOUR
TI - CRISPR/Cas9-mediated loss of FERMT3 reveals kindlin-3-independent neutrophil activity in LAD-III.
AU - Steven D S Webbers
AU - Felipe Rojas-Rodriguez
AU - Angela A F Gankema
AU - Imran Khan
AU - Judy Geissler
AU - Anton T J Tool
AU - Rupert Öllinger
AU - Robin van Bruggen
AU - Teunis J P van Dam
AU - Suzanna C Fagerholm
AU - Markus Moser
AU - Taco W Kuijpers
PY - 2026
DA - 2026/10/21
JO - Journal of human immunity
DO - 10.70962/jhi.20260086
UR - https://europepmc.org/article/MED/42905029
AB - Mutations in human FERMT3 (encoding kindlin-3 protein) impair β2 integrin activity in neutrophils from patients with leukocyte adhesion deficiency III (LAD-III). Yet kindlin-3's contribution to neutrophil integrin affinity regulation and function remains unclear. Using CD34+ human hematopoietic stem and progenitor cells (HSPCs) differentiated into neutrophils, we investigated how CRISPR/Cas9-mediated kindlin-3 deletion affects neutrophil characteristics. CRISPR/Cas9 targeting FERMT3 resulted in an ∼80-bp deletion within exon 2, impairing protein production. FERMT3 KO neutrophils retained normal in vitro development and phenotype with similar surface marker expression, nuclear morphology, and expansion potential compared with untreated cultured neutrophils. Loss of kindlin-3 abolished zymosan-induced NADPH oxidase activity, recapitulating the ROS production defect of LAD-III neutrophils. However, although basal adhesion was reduced, stimulus-induced integrin activity remained largely intact. FERMT3 KO neutrophils retained fMLP-driven chemotaxis while showing impaired migration in response to C5a. Together, these findings show that disruption of kindlin-3 largely recapitulates the LAD-III phenotype while exerting some effector functions upon stimulation without intact kindlin-3.
SN - 3065-8993
ER -