Successful treatment of murine β-thalassemia using in vivo selection of genetically modified, drug-resistant hematopoietic stem cells
Autor: | Thomas P. Brent, Derek A. Persons, Hideki Hanawa, Nobukuni Sawai, Arthur W. Nienhuis, Phillip W. Hargrove, Esther R. Allay, Brian P. Sorrentino |
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Rok vydání: | 2003 |
Předmět: |
Male
Guanine Transplantation Conditioning Recombinant Fusion Proteins medicine.medical_treatment Genetic enhancement Genetic Vectors Immunology Drug Resistance Hematopoietic stem cell transplantation Biology Biochemistry Viral vector Mice O(6)-Methylguanine-DNA Methyltransferase Temozolomide medicine Animals Selection Genetic Crosses Genetic Transplantation Chimera Lentivirus beta-Thalassemia Genetic transfer Hematopoietic Stem Cell Transplantation Cell Biology Hematology Hematopoietic Stem Cells Mice Mutant Strains Globins Dacarbazine Mice Inbred C57BL Haematopoiesis medicine.anatomical_structure Cancer research Erythropoiesis Female Bone marrow Stem cell |
Zdroj: | Blood. 102:506-513 |
ISSN: | 1528-0020 0006-4971 |
DOI: | 10.1182/blood-2003-03-0677 |
Popis: | Successful gene therapy of β-thalassemia will require replacement of the abnormal erythroid compartment with erythropoiesis derived from genetically corrected, autologous hematopoietic stem cells (HSCs). However, currently attainable gene transfer efficiencies into human HSCs are unlikely to yield sufficient numbers of corrected cells for a clinical benefit. Here, using a murine model of β-thalassemia, we demonstrate for the first time that selective enrichment in vivo of transplanted, drug-resistant HSCs can be used therapeutically and may therefore be a useful approach to overcome limiting gene transfer. We used an oncoretroviral vector to transfer a methylguanine methyltransferase (MGMT) drug-resistance gene into normal bone marrow cells. These cells were transplanted into β-thalassemic mice given nonmyeloablative pretransplantation conditioning with temozolomide (TMZ) and O6-benzylguanine (BG). A majority of mice receiving 2 additional courses of TMZ/BG demonstrated in vivo selection of the drug-resistant cells and amelioration of anemia, compared with untreated control animals. These results were extended using a novel γ-globin/MGMT dual gene lentiviral vector. Following drug treatment, normal mice that received transduced cells had an average 67-fold increase in γ-globin expressing red cells. These studies demonstrate that MGMT-based in vivo selection may be useful to increase genetically corrected cells to therapeutic levels in patients with β-thalassemia. |
Databáze: | OpenAIRE |
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