title: Engraftment of immune-deficient mice with primitive hematopoietic cells from beta-thalassemia and sickle cell anemia patients: implications for evaluating human gene therapy protocols.
abstract: Permanent correction of genetic deficiencies of the hematopoietic system requires gene transfer into stem cells and long-term lineage specific expression after autologous transplantation. However, progress to develop gene therapy protocols has been hampered by the absence of in vivo assays that detect genetically deficient human hematopoietic stem cells and their diseased differentiated progeny. The establishment of systems to transplant human cells into immune-deficient SCID mice provides such an assay. We report that primitive bone marrow cells from beta-thalassemia major and sickle cell anemia patients engraft immune-deficient mice, giving rise to high levels of human erythroid and myeloid cells in response to treatment with human cytokines. The bone marrow of transplanted mice contained the entire erythroid lineage from BFU-E to mature erythrocytes expressing human gamma, beta or beta s-globin. Moreover, human erythroid cells from mice transplanted with sickle cell anemia bone marrow showed characteristic sickling under reducing conditions in an in vitro assay. This model provides a powerful in vivo system that can be used to evaluate the efficiency of globin gene transfer into primitive human hematopoietic cells, lineage-specific expression in mature erythrocytes, and ultimately correction of the cellular defect found in the erythroid lineage.
authors.affiliation: Department of Genetics, Hospital for Sick Children, Toronto, Ontario, Canada.
authors.forename: A
authors.initials: A
authors.lastname: Larochelle
authors.forename: J
authors.initials: J
authors.lastname: Vormoor
authors.forename: T
authors.initials: T
authors.lastname: Lapidot
authors.forename: G
authors.initials: G
authors.lastname: Sher
authors.forename: T
authors.initials: T
authors.lastname: Furukawa
authors.forename: Q
authors.initials: Q
authors.lastname: Li
authors.forename: L D
authors.initials: LD
authors.lastname: Shultz
authors.forename: N F
authors.initials: NF
authors.lastname: Olivieri
authors.forename: G
authors.initials: G
authors.lastname: Stamatoyannopoulos
authors.forename: J E
authors.initials: JE
authors.lastname: Dick
mesh_terms: D000755:Anemia, Sickle Cell / Q000235:genetics* / Q000473:pathology / Q000628:therapy; D000818:Animals; D001854:Bone Marrow Cells; D002454:Cell Differentiation; D002455:Cell Division; D002985:Clinical Protocols / Q000592:standards; D004195:Disease Models, Animal; D015672:Erythroid Precursor Cells; D005069:Evaluation Studies as Topic; D005786:Gene Expression Regulation; D015316:Genetic Therapy / Q000592:standards; D005822:Genetic Vectors / Q000592:standards; D005914:Globins / Q000235:genetics / Q000378:metabolism; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D014158:Transcription, Genetic; D017086:beta-Thalassemia / Q000235:genetics* / Q000473:pathology / Q000628:therapy
chemical_list: D005914:Globins