Oxidase-deficient neutrophils from X-linked chronic granulomatous disease iPS cells

Functional correction by zinc finger nuclease-mediated safe harbor targeting

Jizhong Zou, Colin L. Sweeney, Bin Kuan Chou, Uimook Choi, Jason Pan, Hongmei Wang, Sarah N. Dowey, Linzhao Cheng, Harry L. Malech

Research output: Contribution to journalArticle

Abstract

We have developed induced pluripotent stem cells (iPSCs) from a patient with X-linked chronic granulomatous disease (X-CGD), a defect of neutrophil microbicidal reactive oxygen species (ROS) generation resulting from gp91 phox deficiency. We demonstrated that mature neutrophils differentiated from X-CGD iPSCs lack ROS production, reproducing the pathognomonic CGD cellular phenotype. Targeted gene transfer into iPSCs, with subsequent selection and full characterization to ensure no off-target changes, holds promise for correction of monogenic diseases without the insertional mutagenesis caused by multisite integration of viral or plasmid vectors. Zinc finger nuclease-mediated gene targeting of a single-copy gp91phox therapeutic minigene into one allele of the "safe harbor" AAVS1 locus in X-CGD iPSCs without off-target inserts resulted in sustained expression of gp91phox and substantially restored neutrophil ROS production. Our findings demonstrate how precise gene targeting may be applied to correction of X-CGD using zinc finger nuclease and patient iPSCs.

Original languageEnglish (US)
Pages (from-to)5561-5572
Number of pages12
JournalBlood
Volume117
Issue number21
DOIs
StatePublished - May 26 2011

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Chronic Granulomatous Disease
Induced Pluripotent Stem Cells
Zinc Fingers
Ports and harbors
Stem cells
Zinc
Oxidoreductases
Neutrophils
Reactive Oxygen Species
Gene Targeting
Genes
Virus Integration
Gene transfer
Mutagenesis
Insertional Mutagenesis
Plasmids
Alleles
Phenotype
Defects

ASJC Scopus subject areas

  • Hematology
  • Biochemistry
  • Cell Biology
  • Immunology

Cite this

Oxidase-deficient neutrophils from X-linked chronic granulomatous disease iPS cells : Functional correction by zinc finger nuclease-mediated safe harbor targeting. / Zou, Jizhong; Sweeney, Colin L.; Chou, Bin Kuan; Choi, Uimook; Pan, Jason; Wang, Hongmei; Dowey, Sarah N.; Cheng, Linzhao; Malech, Harry L.

In: Blood, Vol. 117, No. 21, 26.05.2011, p. 5561-5572.

Research output: Contribution to journalArticle

Zou, Jizhong ; Sweeney, Colin L. ; Chou, Bin Kuan ; Choi, Uimook ; Pan, Jason ; Wang, Hongmei ; Dowey, Sarah N. ; Cheng, Linzhao ; Malech, Harry L. / Oxidase-deficient neutrophils from X-linked chronic granulomatous disease iPS cells : Functional correction by zinc finger nuclease-mediated safe harbor targeting. In: Blood. 2011 ; Vol. 117, No. 21. pp. 5561-5572.
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