title: Complete genetic correction of ips cells from Duchenne muscular dystrophy.
abstract: Human artificial chromosome (HAC) has several advantages as a gene therapy vector, including stable episomal maintenance that avoids insertional mutations and the ability to carry large gene inserts including the regulatory elements. Induced pluripotent stem (iPS) cells have great potential for gene therapy, as such cells can be generated from the individual's own tissues, and when reintroduced can contribute to the specialized function of any tissue. As a proof of concept, we show herein the complete correction of a genetic deficiency in iPS cells derived from Duchenne muscular dystrophy (DMD) model (mdx) mice and a human DMD patient using a HAC with a complete genomic dystrophin sequence (DYS-HAC). Deletion or mutation of dystrophin in iPS cells was corrected by transferring the DYS-HAC via microcell-mediated chromosome transfer (MMCT). DMD patient- and mdx-specific iPS cells with the DYS-HAC gave rise to differentiation of three germ layers in the teratoma, and human dystrophin expression was detected in muscle-like tissues. Furthermore, chimeric mice from mdx-iPS (DYS-HAC) cells were produced and DYS-HAC was detected in all tissues examined, with tissue-specific expression of dystrophin. Therefore, the combination of patient-specific iPS cells and HAC-containing defective genes represents a powerful tool for gene and cell therapies.
authors.affiliation: Department of Biomedical Science, Institute of Regenerative Medicine and Biofunction, Graduate School of Medical Science, Tottori University, Yonago, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Kazuki
authors.forename: Masaharu
authors.initials: M
authors.lastname: Hiratsuka
authors.forename: Masato
authors.initials: M
authors.lastname: Takiguchi
authors.forename: Mitsuhiko
authors.initials: M
authors.lastname: Osaki
authors.forename: Naoyo
authors.initials: N
authors.lastname: Kajitani
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Hoshiya
authors.forename: Kei
authors.initials: K
authors.lastname: Hiramatsu
authors.forename: Toko
authors.initials: T
authors.lastname: Yoshino
authors.forename: Kanako
authors.initials: K
authors.lastname: Kazuki
authors.forename: Chie
authors.initials: C
authors.lastname: Ishihara
authors.forename: Shoko
authors.initials: S
authors.lastname: Takehara
authors.forename: Katsumi
authors.initials: K
authors.lastname: Higaki
authors.forename: Masato
authors.initials: M
authors.lastname: Nakagawa
authors.forename: Kazutoshi
authors.initials: K
authors.lastname: Takahashi
authors.forename: Shinya
authors.initials: S
authors.lastname: Yamanaka
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Oshimura
mesh_terms: D000818:Animals; D016466:CHO Cells; D002460:Cell Line; D002478:Cells, Cultured; D022222:Chromosomes, Artificial, Human / Q000235:genetics; D006224:Cricetinae; D003412:Cricetulus; D016189:Dystrophin / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D018101:Mice, Inbred mdx; D008962:Models, Theoretical; D020388:Muscular Dystrophy, Duchenne / Q000628:therapy*; D016133:Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D016189:Dystrophin