title: Gene Correction of iPSCs from a Wiskott-Aldrich Syndrome Patient Normalizes the Lymphoid Developmental and Functional Defects.
abstract: Wiskott-Aldrich syndrome (WAS) is an X-linked primary immunodeficiency disease caused by mutations in the gene encoding the WAS protein (WASp). Here, induced pluripotent stem cells (iPSCs) were derived from a WAS patient (WAS-iPSC) and the endogenous chromosomal WAS locus was targeted with a wtWAS-2A-eGFP transgene using zinc finger nucleases (ZFNs) to generate corrected WAS-iPSC (cWAS-iPSC). WASp and GFP were first expressed in the earliest CD34(+)CD43(+)CD45(-) hematopoietic precursor cells and later in all hematopoietic lineages examined. Whereas differentiation to non-lymphoid lineages was readily obtained from WAS-iPSCs, in vitro T lymphopoiesis from WAS-iPSC was deficient with few CD4(+)CD8(+) double-positive and mature CD3(+) T cells obtained. T cell differentiation was restored for cWAS-iPSCs. Similarly, defects in natural killer cell differentiation and function were restored on targeted correction of the WAS locus. These results demonstrate that the defects exhibited by WAS-iPSC-derived lymphoid cells were fully corrected and suggests the potential therapeutic use of gene-corrected WAS-iPSCs.
authors.affiliation: Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center, Houston, TX 77030, USA.
authors.forename: Tamara J
authors.initials: TJ
authors.lastname: Laskowski
authors.affiliation: Laboratory for Experimental Immunology, Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, Ghent 9000, Belgium.
authors.forename: Yasmine
authors.initials: Y
authors.lastname: Van Caeneghem
authors.affiliation: Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center, Houston, TX 77030, USA.
authors.forename: Rasoul
authors.initials: R
authors.lastname: Pourebrahim
authors.affiliation: Department of Medicine and Stem Cell Institute, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Chao
authors.initials: C
authors.lastname: Ma
authors.affiliation: Department of Medicine and Stem Cell Institute, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Zhenya
authors.initials: Z
authors.lastname: Ni
authors.affiliation: Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center, Houston, TX 77030, USA; Differentiation and Cytometry Unit, Hematopoietic Innovative Therapies Division, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT) - Centro de Investigaciones Biomédicas en Red de Enfermedades Raras (CIBERER), Madrid 28040, Spain; Advanced Therapies Mixed Unit, Instituto de Investigación Sanitaria-Fundación Jiménez Díaz (IIS-FJD, UAM), Madrid 28040, Spain.
authors.forename: Zita
authors.initials: Z
authors.lastname: Garate
authors.affiliation: Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center, Houston, TX 77030, USA.
authors.forename: Ana M
authors.initials: AM
authors.lastname: Crane
authors.affiliation: Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center, Houston, TX 77030, USA.
authors.forename: Xuan Shirley
authors.initials: XS
authors.lastname: Li
authors.affiliation: Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center, Houston, TX 77030, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Liao
authors.affiliation: Center for Molecular Imaging, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center, Houston, TX 77030, USA.
authors.forename: Manuel
authors.initials: M
authors.lastname: Gonzalez-Garay
authors.affiliation: Differentiation and Cytometry Unit, Hematopoietic Innovative Therapies Division, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT) - Centro de Investigaciones Biomédicas en Red de Enfermedades Raras (CIBERER), Madrid 28040, Spain; Advanced Therapies Mixed Unit, Instituto de Investigación Sanitaria-Fundación Jiménez Díaz (IIS-FJD, UAM), Madrid 28040, Spain.
authors.forename: Jose Carlos
authors.initials: JC
authors.lastname: Segovia
authors.affiliation: Sangamo BioSciences Inc, Richmond, CA 94804, USA.
authors.forename: David E
authors.initials: DE
authors.lastname: Paschon
authors.affiliation: Sangamo BioSciences Inc, Richmond, CA 94804, USA.
authors.forename: Edward J
authors.initials: EJ
authors.lastname: Rebar
authors.affiliation: Sangamo BioSciences Inc, Richmond, CA 94804, USA.
authors.forename: Michael C
authors.initials: MC
authors.lastname: Holmes
authors.affiliation: Department of Medicine and Stem Cell Institute, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Dan
authors.initials: D
authors.lastname: Kaufman
authors.affiliation: Laboratory for Experimental Immunology, Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, Ghent 9000, Belgium.
authors.forename: Bart
authors.initials: B
authors.lastname: Vandekerckhove
authors.affiliation: Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center, Houston, TX 77030, USA. Electronic address: brian.r.davis@uth.tmc.edu.
authors.forename: Brian R
authors.initials: BR
authors.lastname: Davis
mesh_terms: D015316:Genetic Therapy; D006412:Hematopoietic Stem Cells / Q000378:metabolism / Q000473:pathology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000473:pathology*; D007694:Killer Cells, Natural / Q000378:metabolism; D038041:Lymphopoiesis; D013601:T-Lymphocytes / Q000276:immunology; D014923:Wiskott-Aldrich Syndrome / Q000473:pathology* / Q000628:therapy*; D051316:Wiskott-Aldrich Syndrome Protein / Q000235:genetics
chemical_list: C495767:WAS protein, human; D051316:Wiskott-Aldrich Syndrome Protein
keywords: T cells; genome editing; immune deficiency; induced pluripotent stem cells