Defects in Nicotinamide-adenine Dinucleotide Phosphate Oxidase Genes NOX1 and DUOX2 in Very Early Onset Inflammatory Bowel Disease

Patti Hayes, Sandeep Dhillon, Kim O'Neill, Cornelia Thoeni, Ken Y. Hui, Abdul Elkadri, Conghui H. Guo, Lidija Kovacic, Gabriella Aviello, Luis A. Alvarez, Anne M. Griffiths, Scott B. Snapper, Steven R. Brant, James H. Doroshow, Mark S. Silverberg, Inga Peter, Dermot P B McGovern, Judy Cho, John H. Brumell, Holm H. UhligBilly Bourke, Aleixo M. Muise, Ulla G. Knaus

Research output: Contribution to journalArticle

Abstract

Background & Aims: Defects in intestinal innate defense systems predispose patients to inflammatory bowel disease (IBD). Reactive oxygen species (ROS) generated by nicotinamide-adenine dinucleotide phosphate (NADPH) oxidases in the mucosal barrier maintain gut homeostasis and defend against pathogenic attack. We hypothesized that molecular genetic defects in intestinal NADPH oxidases might be present in children with IBD. Methods: After targeted exome sequencing of epithelial NADPH oxidases NOX1 and DUOX2 on 59 children with very early onset inflammatory bowel disease (VEOIBD), the identified mutations were validated using Sanger Sequencing. A structural analysis of NOX1 and DUOX2 variants was performed by homology in silico modeling. The functional characterization included ROS generation in model cell lines and in invivo transduced murine crypts, protein expression, intracellular localization, and cell-based infection studies with the enteric pathogens Campylobacter jejuni and enteropathogenic Escherichia coli. Results: We identified missense mutations in NOX1 (c.988G>A, p.Pro330Ser; c.967G>A, p.Asp360Asn) and DUOX2 (c.4474G>A, p.Arg1211Cys; c.3631C>T, p.Arg1492Cys) in 5 of 209 VEOIBD patients. The NOX1 p.Asp360Asn variant was replicated in a male Ashkenazi Jewish ulcerative colitis cohort. Patients with both NOX1 and DUOX2 variants showed abnormal Paneth cell metaplasia. All NOX1 and DUOX2 variants showed reduced ROS production compared with wild-type enzymes. Despite appropriate cellular localization and comparable pathogen-stimulated translocation of altered oxidases, cells harboring NOX1 or DUOX2 variants had defective host resistance to infection with C.jejuni. Conclusions: This study identifies the first inactivating missense variants in NOX1 and DUOX2 associated with VEOIBD. Defective ROS production from intestinal epithelial cells constitutes a risk factor for developing VEOIBD.

Original languageEnglish (US)
Pages (from-to)489-502
Number of pages14
JournalCMGH Cellular and Molecular Gastroenterology and Hepatology
Volume1
Issue number5
DOIs
StatePublished - Sep 1 2015

Fingerprint

NADP
Inflammatory Bowel Diseases
Oxidoreductases
Reactive Oxygen Species
Genes
Paneth Cells
Exome
Enteropathogenic Escherichia coli
Campylobacter jejuni
Metaplasia
Missense Mutation
Infection
Ulcerative Colitis
Computer Simulation
Molecular Biology
Homeostasis
Epithelial Cells
Cell Line
Mutation
Enzymes

Keywords

  • DUOX2
  • Inflammatory Bowel Disease
  • NADPH Oxidase
  • NOX1
  • Reactive Oxygen Species
  • VEOIBD

ASJC Scopus subject areas

  • Gastroenterology
  • Hepatology

Cite this

Defects in Nicotinamide-adenine Dinucleotide Phosphate Oxidase Genes NOX1 and DUOX2 in Very Early Onset Inflammatory Bowel Disease. / Hayes, Patti; Dhillon, Sandeep; O'Neill, Kim; Thoeni, Cornelia; Hui, Ken Y.; Elkadri, Abdul; Guo, Conghui H.; Kovacic, Lidija; Aviello, Gabriella; Alvarez, Luis A.; Griffiths, Anne M.; Snapper, Scott B.; Brant, Steven R.; Doroshow, James H.; Silverberg, Mark S.; Peter, Inga; McGovern, Dermot P B; Cho, Judy; Brumell, John H.; Uhlig, Holm H.; Bourke, Billy; Muise, Aleixo M.; Knaus, Ulla G.

In: CMGH Cellular and Molecular Gastroenterology and Hepatology, Vol. 1, No. 5, 01.09.2015, p. 489-502.

Research output: Contribution to journalArticle

Hayes, P, Dhillon, S, O'Neill, K, Thoeni, C, Hui, KY, Elkadri, A, Guo, CH, Kovacic, L, Aviello, G, Alvarez, LA, Griffiths, AM, Snapper, SB, Brant, SR, Doroshow, JH, Silverberg, MS, Peter, I, McGovern, DPB, Cho, J, Brumell, JH, Uhlig, HH, Bourke, B, Muise, AM & Knaus, UG 2015, 'Defects in Nicotinamide-adenine Dinucleotide Phosphate Oxidase Genes NOX1 and DUOX2 in Very Early Onset Inflammatory Bowel Disease', CMGH Cellular and Molecular Gastroenterology and Hepatology, vol. 1, no. 5, pp. 489-502. https://doi.org/10.1016/j.jcmgh.2015.06.005
Hayes, Patti ; Dhillon, Sandeep ; O'Neill, Kim ; Thoeni, Cornelia ; Hui, Ken Y. ; Elkadri, Abdul ; Guo, Conghui H. ; Kovacic, Lidija ; Aviello, Gabriella ; Alvarez, Luis A. ; Griffiths, Anne M. ; Snapper, Scott B. ; Brant, Steven R. ; Doroshow, James H. ; Silverberg, Mark S. ; Peter, Inga ; McGovern, Dermot P B ; Cho, Judy ; Brumell, John H. ; Uhlig, Holm H. ; Bourke, Billy ; Muise, Aleixo M. ; Knaus, Ulla G. / Defects in Nicotinamide-adenine Dinucleotide Phosphate Oxidase Genes NOX1 and DUOX2 in Very Early Onset Inflammatory Bowel Disease. In: CMGH Cellular and Molecular Gastroenterology and Hepatology. 2015 ; Vol. 1, No. 5. pp. 489-502.
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T1 - Defects in Nicotinamide-adenine Dinucleotide Phosphate Oxidase Genes NOX1 and DUOX2 in Very Early Onset Inflammatory Bowel Disease

AU - Hayes, Patti

AU - Dhillon, Sandeep

AU - O'Neill, Kim

AU - Thoeni, Cornelia

AU - Hui, Ken Y.

AU - Elkadri, Abdul

AU - Guo, Conghui H.

AU - Kovacic, Lidija

AU - Aviello, Gabriella

AU - Alvarez, Luis A.

AU - Griffiths, Anne M.

AU - Snapper, Scott B.

AU - Brant, Steven R.

AU - Doroshow, James H.

AU - Silverberg, Mark S.

AU - Peter, Inga

AU - McGovern, Dermot P B

AU - Cho, Judy

AU - Brumell, John H.

AU - Uhlig, Holm H.

AU - Bourke, Billy

AU - Muise, Aleixo M.

AU - Knaus, Ulla G.

PY - 2015/9/1

Y1 - 2015/9/1

N2 - Background & Aims: Defects in intestinal innate defense systems predispose patients to inflammatory bowel disease (IBD). Reactive oxygen species (ROS) generated by nicotinamide-adenine dinucleotide phosphate (NADPH) oxidases in the mucosal barrier maintain gut homeostasis and defend against pathogenic attack. We hypothesized that molecular genetic defects in intestinal NADPH oxidases might be present in children with IBD. Methods: After targeted exome sequencing of epithelial NADPH oxidases NOX1 and DUOX2 on 59 children with very early onset inflammatory bowel disease (VEOIBD), the identified mutations were validated using Sanger Sequencing. A structural analysis of NOX1 and DUOX2 variants was performed by homology in silico modeling. The functional characterization included ROS generation in model cell lines and in invivo transduced murine crypts, protein expression, intracellular localization, and cell-based infection studies with the enteric pathogens Campylobacter jejuni and enteropathogenic Escherichia coli. Results: We identified missense mutations in NOX1 (c.988G>A, p.Pro330Ser; c.967G>A, p.Asp360Asn) and DUOX2 (c.4474G>A, p.Arg1211Cys; c.3631C>T, p.Arg1492Cys) in 5 of 209 VEOIBD patients. The NOX1 p.Asp360Asn variant was replicated in a male Ashkenazi Jewish ulcerative colitis cohort. Patients with both NOX1 and DUOX2 variants showed abnormal Paneth cell metaplasia. All NOX1 and DUOX2 variants showed reduced ROS production compared with wild-type enzymes. Despite appropriate cellular localization and comparable pathogen-stimulated translocation of altered oxidases, cells harboring NOX1 or DUOX2 variants had defective host resistance to infection with C.jejuni. Conclusions: This study identifies the first inactivating missense variants in NOX1 and DUOX2 associated with VEOIBD. Defective ROS production from intestinal epithelial cells constitutes a risk factor for developing VEOIBD.

AB - Background & Aims: Defects in intestinal innate defense systems predispose patients to inflammatory bowel disease (IBD). Reactive oxygen species (ROS) generated by nicotinamide-adenine dinucleotide phosphate (NADPH) oxidases in the mucosal barrier maintain gut homeostasis and defend against pathogenic attack. We hypothesized that molecular genetic defects in intestinal NADPH oxidases might be present in children with IBD. Methods: After targeted exome sequencing of epithelial NADPH oxidases NOX1 and DUOX2 on 59 children with very early onset inflammatory bowel disease (VEOIBD), the identified mutations were validated using Sanger Sequencing. A structural analysis of NOX1 and DUOX2 variants was performed by homology in silico modeling. The functional characterization included ROS generation in model cell lines and in invivo transduced murine crypts, protein expression, intracellular localization, and cell-based infection studies with the enteric pathogens Campylobacter jejuni and enteropathogenic Escherichia coli. Results: We identified missense mutations in NOX1 (c.988G>A, p.Pro330Ser; c.967G>A, p.Asp360Asn) and DUOX2 (c.4474G>A, p.Arg1211Cys; c.3631C>T, p.Arg1492Cys) in 5 of 209 VEOIBD patients. The NOX1 p.Asp360Asn variant was replicated in a male Ashkenazi Jewish ulcerative colitis cohort. Patients with both NOX1 and DUOX2 variants showed abnormal Paneth cell metaplasia. All NOX1 and DUOX2 variants showed reduced ROS production compared with wild-type enzymes. Despite appropriate cellular localization and comparable pathogen-stimulated translocation of altered oxidases, cells harboring NOX1 or DUOX2 variants had defective host resistance to infection with C.jejuni. Conclusions: This study identifies the first inactivating missense variants in NOX1 and DUOX2 associated with VEOIBD. Defective ROS production from intestinal epithelial cells constitutes a risk factor for developing VEOIBD.

KW - DUOX2

KW - Inflammatory Bowel Disease

KW - NADPH Oxidase

KW - NOX1

KW - Reactive Oxygen Species

KW - VEOIBD

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