NOX1 Regulates Collective and Planktonic Cell Migration: Insights From Patients With Pediatric-Onset IBD and NOX1 Deficiency
| Autor: | Razieh Khoshnevisan, Roya Sherkat, Katrin Schröder, Franziska Moll, Michael W. Anderson, Christoph Walz, Scott B. Snapper, Sebastian Hollizeck, Alireza Andalib, Meino Rohlfs, Daniel Kotlarz, Sierra Anderson, Christoph Klein, Abbas Rezaei, David Illig, Peyman Adibi, S. Koletzko, Stephen Babcock, Benjamin Marquardt, Jay R. Thiagarajah, Aleixo M. Muise |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Adult Male Adolescent Cell Biology medicine.disease_cause Cell Line Focal adhesion 03 medical and health sciences Young Adult 0302 clinical medicine Epidermal growth factor Cell Movement medicine Immunology and Allergy Humans Intestinal Mucosa Child Mutation Wound Healing Gastroenterology Cell migration Chemotaxis Inflammatory Bowel Diseases Cytoskeletal Proteins 030104 developmental biology medicine.anatomical_structure Cell culture 030220 oncology & carcinogenesis Cancer research cardiovascular system NADPH Oxidase 1 Female Wound healing Basic Science Research |
| Zdroj: | Inflamm Bowel Dis |
| Popis: | Background Genetic defects of pediatric-onset inflammatory bowel disease (IBD) provide critical insights into molecular factors controlling intestinal homeostasis. NOX1 has been recently recognized as a major source of reactive oxygen species (ROS) in human colonic epithelial cells. Here we assessed the functional consequences of human NOX1 deficiency with respect to wound healing and epithelial migration by studying pediatric IBD patients presenting with a stop-gain mutation in NOX1. Methods Functional characterization of the NOX1 variant included ROS generation, wound healing, 2-dimensional collective chemotactic migration, single-cell planktonic migration in heterologous cell lines, and RNA scope and immunohistochemistry of paraffin-embedded patient tissue samples. Results Using exome sequencing, we identified a stop-gain mutation in NOX1 (c.160C>T, p.54R>*) in patients with pediatric-onset IBD. Our studies confirmed that loss-of-function of NOX1 causes abrogated ROS activity, but they also provided novel mechanistic insights into human NOX1 deficiency. Cells that were NOX1-mutant showed impaired wound healing and attenuated 2-dimensional collective chemotactic migration. High-resolution microscopy of the migrating cell edge revealed a reduced density of filopodial protrusions with altered focal adhesions in NOX1-deficient cells, accompanied by reduced phosphorylation of p190A. Assessment of single-cell planktonic migration toward an epidermal growth factor gradient showed that NOX1 deficiency is associated with altered migration dynamics with loss of directionality and altered cell-cell interactions. Conclusions Our studies on pediatric-onset IBD patients with a rare sequence variant in NOX1 highlight that human NOX1 is involved in regulating wound healing by altering epithelial cytoskeletal dynamics at the leading edge and directing cell migration. |
| Databáze: | OpenAIRE |
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