title: Recombinant AAV2 transduction of primitive human hematopoietic stem cells capable of serial engraftment in immune-deficient mice.
abstract: A recombinant AAV2 (rAAV2) vector encoding antisense RNA to HIV-1 transactivating region (TAR) was evaluated for transduction of human cord blood CD34+CD38- hematopoietic stem cells (HSC) capable of serial engraftment in nonobese diabetic (NOD)/severe combined immunodeficient (SCID) mice. Results revealed long-term multilineage marking in primary and secondary recipients, and significantly, an enrichment of transduced cells in secondary hosts, indicating efficient transduction of multipotential self-renewing HSC. These results were confirmed by the persistence of rAAV marking of clonogenic progenitors in serial analyses of recipient marrow. Upon HIV-1 challenge, the macrophage progeny of transduced CD34+ cells expressed antisense RNA and exhibited sustained and significant inhibition of virus replication as compared with controls in every donor tested, without selective pressure. This study represents a clear in vivo demonstration of efficient rAAV2 transduction of human HSC.
authors.affiliation: Divisions of Virology and Hematology/Hematopoietic Cell Transplantation, City of Hope National Medical Center, Duarte, CA 91010, USA.
authors.forename: Leah
authors.initials: L
authors.lastname: Santat
authors.forename: Helicia
authors.initials: H
authors.lastname: Paz
authors.forename: Christie
authors.initials: C
authors.lastname: Wong
authors.forename: Lijing
authors.initials: L
authors.lastname: Li
authors.forename: James
authors.initials: J
authors.lastname: Macer
authors.forename: Stephen
authors.initials: S
authors.lastname: Forman
authors.forename: K K
authors.initials: KK
authors.lastname: Wong
authors.forename: Saswati
authors.initials: S
authors.lastname: Chatterjee
mesh_terms: D036541:ADP-ribosyl Cyclase / Q000378:metabolism; D051997:ADP-ribosyl Cyclase 1; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D018952:Antigens, CD34 / Q000378:metabolism; D003114:Colony-Forming Units Assay; D000229:Dependovirus / Q000235:genetics; D005312:Fetal Blood / Q000166:cytology / Q000276:immunology; D005822:Genetic Vectors; D006085:Graft Survival; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000235:genetics*; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D008297:Male; D008562:Membrane Glycoproteins; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D016372:RNA, Antisense / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D014161:Transduction, Genetic
chemical_list: D015703:Antigens, CD; D018952:Antigens, CD34; D008562:Membrane Glycoproteins; D016372:RNA, Antisense; D012367:RNA, Viral; D036541:ADP-ribosyl Cyclase; C498154:CD38 protein, human; C498155:Cd38 protein, mouse; D051997:ADP-ribosyl Cyclase 1