title: Therapeutic approaches for treating hemophilia A using embryonic stem cells.
abstract: Hemophilia A is an X-linked rescessive bleeding disorder that results from F8 gene aberrations. Previously, we established embryonic stem (ES) cells (tet-226aa/N6-Ainv18) that secrete human factor VIII (hFVIII) by introducing the human F8 gene in mouse Ainv18 ES cells. Here, we explored the potential of cell transplantation therapy for hemophilia A using the ES cells. Transplant tet-226aa/N6-Ainv18 ES cells were injected into the spleens of severe combined immunodeficiency (SCID) mice, carbon tetrachloride (CCl4)-pretreated wild-type mice, and CCl4-pretreated hemophilia A mice. F8 expression was induced by doxycycline in drinking water, and hFVIII-antigen production was assessed in all cell transplantation experiments. Injecting the ES cells into SCID mice resulted in an enhanced expression of the hFVIII antigen; however, teratoma generation was confirmed in the spleen. Transplantation of ES cells into wild-type mice after CCl4-induced liver injury facilitated survival and engraftment of transplanted cells without teratoma formation, resulting in hFVIII production in the plasma. Although CCl4 was lethal to most hemophilia A mice, therapeutic levels of FVIII activity, as well as the hFVIII antigen, were detected in surviving hemophilia A mice after cell transplantation. Immunolocalization results for hFVIII suggested that transplanted ES cells might be engrafted at the periportal area in the liver. Although the development of a safer induction method for liver regeneration is required, our results suggested the potential for developing an effective ES-cell transplantation therapeutic model for treating hemophilia A in the future.
authors.affiliation: Department of Legal Medicine, Nara Medical University School of Medicine, Kashihara, Japan.
authors.forename: Shogo
authors.initials: S
authors.lastname: Kasuda
authors.affiliation: Department of Pediatrics, Nara Medical University School of Medicine, Kashihara, Japan.
authors.forename: Kohei
authors.initials: K
authors.lastname: Tatsumi
authors.affiliation: Department of Pediatrics, Nara Medical University School of Medicine, Kashihara, Japan; Department of Pediatrics, Matsubara Tokushukai Hospital, Matsubara, Japan. Electronic address: ysakurai-th@umin.ac.jp.
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Sakurai
authors.affiliation: Department of Pediatrics, Nara Medical University School of Medicine, Kashihara, Japan.
authors.forename: Midori
authors.initials: M
authors.lastname: Shima
authors.affiliation: Department of Legal Medicine, Nara Medical University School of Medicine, Kashihara, Japan.
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Hatake
mesh_terms: D000818:Animals; D001777:Blood Coagulation; D002251:Carbon Tetrachloride; D005169:Factor VIII / Q000378:metabolism; D006467:Hemophilia A / Q000628:therapy*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D016513:Mice, SCID; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology*; D033581:Stem Cell Transplantation
chemical_list: C078147:F8 protein, human; D005169:Factor VIII; D002251:Carbon Tetrachloride
keywords: Cell therapy; Embryonic stem cell; Hemophilia A; Mouse model