A congenital neutrophil defect syndrome associated with mutations in VPS45

  • N Engl J Med. 2013 Jul 4;369(1):54-65. doi: 10.1056/NEJMoa1301296.
Thierry Vilboux  1 ,  Atar Lev ,  May Christine V Malicdan ,  Amos J Simon ,  Päivi Järvinen ,  Tomas Racek ,  Jacek Puchalka ,  Raman Sood ,  Blake Carrington ,  Kevin Bishop ,  James Mullikin ,  Marjan Huizing ,  Ben Zion Garty ,  Eran Eyal ,  Baruch Wolach ,  Ronit Gavrieli ,  Amos Toren ,  Michalle Soudack ,  Osama M Atawneh ,  Tatiana Babushkin ,  Ginette Schiby ,  Andrew Cullinane ,  Camila Avivi ,  Sylvie Polak-Charcon ,  Iris Barshack ,  Ninette Amariglio ,  Gideon Rechavi ,  Jutte van der Werff ten Bosch ,  Yair Anikster ,  Christoph Klein ,  William A Gahl ,  Raz Somech
Affiliations
  • 1. Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, USA.
PMID: 23738510 DOI: 10.1056/NEJMoa1301296
Abstract

Background: Neutrophils are the predominant phagocytes that provide protection against Bacterial and Fungal infections. Genetically determined neutrophil disorders confer a predisposition to severe infections and reveal novel mechanisms that control vesicular trafficking, hematopoiesis, and innate immunity.

Methods: We clinically evaluated seven children from five families who had neutropenia, neutrophil dysfunction, bone marrow fibrosis, and nephromegaly. To identify the causative gene, we performed homozygosity mapping using single-nucleotide polymorphism arrays, whole-exome Sequencing, immunoblotting, immunofluorescence, electron microscopy, a real-time quantitative polymerase-chain-reaction assay, immunohistochemistry, flow cytometry, fibroblast motility assays, measurements of Apoptosis, and zebrafish models. Correction experiments were performed by transfecting mutant fibroblasts with the nonmutated gene.

Results: All seven affected children had homozygous mutations (Thr224Asn or Glu238Lys, depending on the child's ethnic origin) in VPS45, which encodes a protein that regulates membrane trafficking through the endosomal system. The level of VPS45 protein was reduced, as were the VPS45 binding partners rabenosyn-5 and syntaxin-16. The level of β1 Integrin was reduced on the surface of VPS45-deficient neutrophils and fibroblasts. VPS45-deficient fibroblasts were characterized by impaired motility and increased Apoptosis. A zebrafish model of vps45 deficiency showed a marked paucity of myeloperoxidase-positive cells (i.e., neutrophils). Transfection of patient cells with nonmutated VPS45 corrected the migration defect and decreased Apoptosis.

Conclusions: Defective endosomal intracellular protein trafficking due to biallelic mutations in VPS45 underlies a new immunodeficiency syndrome involving impaired neutrophil function. (Funded by the National Human Genome Research Institute and Others.).