title: An induced pluripotent stem cell-mediated and integration-free factor VIII expression system.
abstract: Human artificial chromosome (HAC) has several advantages as a gene therapy vector, including stable episomal maintenance and the ability to carry large gene inserts. Induced pluripotent stem (iPS) cells also have a great potential for gene therapy, which can be generated from an individual's own tissues and contribute to any tissues when reintroduced. A Sendai virus (SeV) vector with reprogramming factors is a powerful tool for generating iPS cells because of the high infection efficiency without the risk of integration into host chromosomes. In this study, we developed an iPS cell-mediated and integration-free coagulation factor VIII (FVIII) expression system using non-integrating SeV- and HAC-vectors. Multiple human FVIII genes, which were under the control of the megakaryocyte-specific platelet factor-4 (PF4) promoter for development of a treatment for hemophilia A, were inserted into a HAC vector (PF4-FVIII-HAC). The PF4-FVIII-HAC was introduced into SeV vector-mediated iPS cells derived from a mouse model of hemophilia A. After in vitro differentiation of iPS cells with the PF4-FVIII-HAC into megakaryocytes/platelets, the PF4-FVIII-HAC resulted in expression of FVIII. This study has developed the iPS cell-mediated PF4-driven FVIII expression system using two non-integrating vectors; therefore, this system may be a promising tool for safer gene- and cell-therapy of hemophilia A.
authors.affiliation: Department of Biomedical Science, Institute of Regenerative Medicine and Biofunction, Graduate School of Medical Science, Tottori University, Tottori, Japan.
authors.forename: Yuwna
authors.initials: Y
authors.lastname: Yakura
authors.forename: Chie
authors.initials: C
authors.lastname: Ishihara
authors.forename: Hajime
authors.initials: H
authors.lastname: Kurosaki
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Kazuki
authors.forename: Norio
authors.initials: N
authors.lastname: Komatsu
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Okada
authors.forename: Takefumi
authors.initials: T
authors.lastname: Doi
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Takeya
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Oshimura
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D022222:Chromosomes, Artificial, Human / Q000235:genetics*; D004195:Disease Models, Animal; D005169:Factor VIII / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006467:Hemophilia A / Q000628:therapy*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism* / Q000637:transplantation*; D051379:Mice; D010978:Platelet Factor 4 / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D029082:Sendai virus
chemical_list: D010978:Platelet Factor 4; D005169:Factor VIII