title: Long-term genetic modification of rhesus monkey hematopoietic cells following transplantation of adenoassociated virus vector-transduced CD34+ cells.
abstract: We have explored the potential of recombinant adenoassociated virus (AAV) vectors for gene transfer of the human beta-globin gene and the genetic modification of primate pluripotent hematopoietic stem cells (P-PHSCs). Transduction of P-PHSCs was tested in a preclinical bone marrow transplantation model in rhesus monkeys. CD34+ cells were transduced ex vivo and autologously transplanted without prior selection into irradiated rhesus monkeys. Vector-transduced peripheral blood mononuclear cells and granulocytes were present in the circulation for more than 15 months after transplantation. Approximately 1 in 10(5) cells in the circulation was vector modified. The vector was detected in the bone marrow, in granulocytes, and in purified populations of B and T cells, thus demonstrating multilineage repopulation by vector-transduced stem cells. Comparison of transduction protocols suggested that short-term culture of P-PHSCs enhances transduction and subsequent repopulation by rAAV-transduced cells. These results demonstrate that rAAV vectors can be used for the permanent genetic modification of a rhesus monkey hematopoietic system in the absence of selective pressure.
authors.affiliation: IntroGene BV, Leiden, The Netherlands.
authors.forename: S
authors.initials: S
authors.lastname: Schimmenti
authors.forename: J
authors.initials: J
authors.lastname: Boesen
authors.forename: E A
authors.initials: EA
authors.lastname: Claassen
authors.forename: D
authors.initials: D
authors.lastname: Valerio
authors.forename: M P
authors.initials: MP
authors.lastname: Einerhand
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000235:genetics / Q000276:immunology*; D016026:Bone Marrow Transplantation; D000229:Dependovirus / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D005914:Globins / Q000096:biosynthesis / Q000235:genetics*; D006098:Granulocytes / Q000276:immunology / Q000821:virology; D006412:Hematopoietic Stem Cells / Q000276:immunology* / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000821:virology; D008253:Macaca mulatta; D016133:Polymerase Chain Reaction / Q000379:methods
chemical_list: D018952:Antigens, CD34; D005914:Globins