Development of new gene therapy for chronic granulomatous disease using Piggyback transposon

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We successfully generated induced pluripotent stem cells (iPSCs) from peripheral T cells of chronic granulomatous disease patient with CYBB gene mutation, using sendai virus vector carrying reprogramming factors OCT3/4, SOX2, KLF4, and c-MYC. In the same way, we generated corrected iPSCs derived from CYBB-transduced T cell using PiggyBac transposon.We confirmed expression of pluripotency markers (Oct4, SSEA-3, SSEA-4, TRA-1-60, TRA-1-81, Nanog) by immunofluorescence assays. In addition, these iPSCs showed teratoma formation when injected into immunodeficient mice. These colonies could be differentiated to monocytes and neutrophils with AGM-3 stroma cells. Monocytes and neutrophils derived from non-corrected and corrected iPSCs were devoid of oxidase activity by DHR.These data suggest iPSCs modified by this method were difficult to maintain transgene expression.