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Mol Ther Methods Clin Dev. 2015 Dec 9;2:15046. doi: 10.1038/mtm.2015.46. eCollection 2015.

An assessment of the effects of ectopic gp91phox expression in XCGD iPSC-derived neutrophils.

Author information

1
Division of Stem Cell Therapy, Center for Stem Cell Biology and Regenerative Medicine, Institute of Medical Science, University of Tokyo , Tokyo, Japan ; Division of Stem Cell Processing/Stem Cell Bank, Center for Stem Cell Biology and Regenerative Medicine, Institute of Medical Science, University of Tokyo , Tokyo, Japan.
2
Japan Science Technology Agency, Exploratory Research for Advanced Technology Nakauchi Stem Cell and Organ Regeneration Project , Tokyo, Japan.
3
Division of Stem Cell Therapy, Center for Stem Cell Biology and Regenerative Medicine, Institute of Medical Science, University of Tokyo , Tokyo, Japan ; Japan Science Technology Agency, Exploratory Research for Advanced Technology Nakauchi Stem Cell and Organ Regeneration Project , Tokyo, Japan.
4
Department of Pediatrics, School of Medicine, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University , Kanazawa, Japan.
5
Research Center for Stem Cell Engineering, National Institute of Advanced, Industrial Science and Technology , Tsukuba, Japan ; Laboratory of Gene Regulation, Faculty of Medicine, University of Tsukuba , Tsukuba, Japan.
6
Research Center for Stem Cell Engineering, National Institute of Advanced, Industrial Science and Technology , Tsukuba, Japan.

Abstract

For the treatment of monogenetic hematological disorders, restoration of transgene expression in affected cell populations is generally considered to have beneficial effects. However, X-linked chronic granulomatous disease (XCGD) is unique since the appearance of functional neutrophils in the peripheral blood following hematopoietic stem cell gene therapy is transient only. One contributing factor could be the occurrence of detrimental effects secondary to ectopic gp91phox expression in neutrophils, which has not been formally demonstrated previously. This study uses iPSCs to model XCGD, which allows the process of differentiation to be studied intensely in vitro. Alpharetroviral vectors carrying a ubiquitous promoter were used to drive the "ectopic" expression of codon optimized gp91phox cDNA. In the mature fraction of neutrophils differentiated from transduced XCGD-iPSCs, cellular recovery in terms of gp91phox expression and reactive oxygen species production was abruptly lost before cells had fully differentiated. Most critically, ectopic gp91phox expression could be identified clearly in the developing fraction of the transduced groups, which appeared to correspond with reduced cell viability. It is possible that this impedes further differentiation of developing neutrophils. Therefore, affording cellular protection from the detrimental effects of ectopic gp91phox expression may improve XCGD clinical outcomes.

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