title: Correction of phenotype in a thalassemia mouse model using a nonmyeloablative marrow transplantation regimen.
abstract: Gene therapy, the replacement of normal human beta- or gamma-globin genes into the hematopoietic stem cells of patients with homozygous beta-thalassemia, is a promising therapy for the future. High-level lineage-specific stable globin expression in transduced cells reinfused into patients in an autologous transplantation setting could be curative, if successful. Previous studies have shown high-level donor chimerism in nonmyeloablated non-thalassemic hosts. We have now studied the conditions for stable long-term engraftment of normal cells into a thalassemia mouse model that lead to high-level donor chimerism and correction of the abnormal phenotype. Thalassemic female mice treated with 0 to 300 cGy whole-body irradiation received transplantations of donor cells harvested from wild-type males. Engraftment of male cells was quantitated by Y-chromosome polymerase chain reaction analysis of blood and marrow progenitors, and changes in hemoglobin levels, red cell morphology, and spleen size were measured at various times posttransplantation. High-level stable donor cell engraftment was achieved in mice given 200 cGy and receiving transplants of 2 x 10(7) or more donor cells. The anemia, abnormal peripheral blood smears, and splenomegaly improved in the thalassemic mice that had successful engraftment. These studies demonstrate that stable and successful levels of engraftment of normal cells can correct the thalassemic phenotype without fully myeloablating the host. This animal model should allow us to test the amount of cytoreduction required and the level of engraftment and beta-globin expression needed in autologous transplantation of beta-globin gene-transduced cells to correct the abnormal phenotype in thalassemic mice, and it may be relevant to human clinical trials, as well.
authors.affiliation: Department of Pediatrics, Columbia University, New York, New York 10032, USA.
authors.forename: M Brigid
authors.initials: MB
authors.lastname: Bradley
authors.forename: Rose M
authors.initials: RM
authors.lastname: Sattler
authors.forename: Harry
authors.initials: H
authors.lastname: Raftopoulos
authors.forename: Maureen
authors.initials: M
authors.lastname: Ward
authors.forename: I Robert
authors.initials: IR
authors.lastname: Grossman
authors.forename: Tim M
authors.initials: TM
authors.lastname: Townes
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Ryan
authors.forename: Arthur
authors.initials: A
authors.lastname: Bank
mesh_terms: D000818:Animals; D016026:Bone Marrow Transplantation / Q000379:methods*; D004247:DNA / Q000097:blood; D004195:Disease Models, Animal; D005260:Female; D015316:Genetic Therapy / Q000379:methods; D005914:Globins / Q000235:genetics; D006085:Graft Survival; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D010641:Phenotype; D013789:Thalassemia / Q000628:therapy*; D018183:Transplantation Chimera; D019172:Transplantation Conditioning / Q000379:methods*; D016896:Treatment Outcome; D014916:Whole-Body Irradiation
chemical_list: D005914:Globins; D004247:DNA