Complete genetic correction of ips cells from Duchenne muscular dystrophy.
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.
Kazuki
Yasuhiro
Y
Department of Biomedical Science, Institute of Regenerative Medicine and Biofunction, Graduate School of Medical Science, Tottori University, Yonago, Japan.
Hiratsuka
Masaharu
M
Takiguchi
Masato
M
Osaki
Mitsuhiko
M
Kajitani
Naoyo
N
Hoshiya
Hidetoshi
H
Hiramatsu
Kei
K
Yoshino
Toko
T
Kazuki
Kanako
K
Ishihara
Chie
C
Takehara
Shoko
S
Higaki
Katsumi
K
Nakagawa
Masato
M
Takahashi
Kazutoshi
K
Yamanaka
Shinya
S
Oshimura
Mitsuo
M
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
D016189:Dystrophin