title: In vivo transduction of primitive mobilized hematopoietic stem cells after intravenous injection of integrating adenovirus vectors.
abstract: Current protocols for hematopoietic stem/progenitor cell (HSPC) gene therapy, involving the transplantation of ex vivo genetically modified HSPCs are complex and not without risk for the patient. We developed a new approach for in vivo HSPC transduction that does not require myeloablation and transplantation. It involves subcutaneous injections of granulocyte-colony-stimulating factor/AMD3100 to mobilize HSPCs from the bone marrow (BM) into the peripheral blood stream and the IV injection of an integrating, helper-dependent adenovirus (HD-Ad5/35++) vector system. These vectors target CD46, a receptor that is uniformly expressed on HSPCs. We demonstrated in human CD46 transgenic mice and immunodeficient mice with engrafted human CD34+cells that HSPCs transduced in the periphery home back to the BM where they stably express the transgene. In hCD46 transgenic mice, we showed that our in vivo HSPC transduction approach allows for the stable transduction of primitive HSPCs. Twenty weeks after in vivo transduction, green fluorescent protein (GFP) marking in BM HSPCs (Lin-Sca1+Kit-cells) in most of the mice was in the range of 5% to 10%. The percentage of GFP-expressing primitive HSPCs capable of forming multilineage progenitor colonies (colony-forming units [CFUs]) increased from 4% of all CFUs at week 4 to 16% at week 12, indicating transduction and expansion of long-term surviving HSPCs. Our approach was well tolerated, did not result in significant transduction of nonhematopoietic tissues, and was not associated with genotoxicty. The ability to stably genetically modify HSPCs without the need of myeloablative conditioning is relevant for a broader clinical application of gene therapy.
authors.affiliation: Division of Medical Genetics, Department of Medicine, University of Washington, Seattle, WA.
authors.forename: Maximilian
authors.initials: M
authors.lastname: Richter
authors.affiliation: Division of Medical Genetics, Department of Medicine, University of Washington, Seattle, WA.
authors.forename: Kamola
authors.initials: K
authors.lastname: Saydaminova
authors.affiliation: Division of Medical Genetics, Department of Medicine, University of Washington, Seattle, WA.
authors.forename: Roma
authors.initials: R
authors.lastname: Yumul
authors.affiliation: Division of Medical Genetics, Department of Medicine, University of Washington, Seattle, WA.
authors.forename: Rohini
authors.initials: R
authors.lastname: Krishnan
authors.affiliation: Institute for Virology and Microbiology, University Witten/Herdecke, Witten, Germany.
authors.forename: Jing
authors.initials: J
authors.lastname: Liu
authors.affiliation: Mobile DNA Group, Max-Delbrück Center for Molecular Medicine, Berlin, Germany.
authors.forename: Eniko-Eva
authors.initials: EE
authors.lastname: Nagy
authors.affiliation: Mobile DNA Group, Max-Delbrück Center for Molecular Medicine, Berlin, Germany.
authors.forename: Manvendra
authors.initials: M
authors.lastname: Singh
authors.affiliation: Mobile DNA Group, Max-Delbrück Center for Molecular Medicine, Berlin, Germany.
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Izsvák
authors.affiliation: Department of Molecular Medicine, The Mayo Clinic, Rochester, MN.
authors.forename: Roberto
authors.initials: R
authors.lastname: Cattaneo
authors.affiliation: Mobile DNA Group, Max-Delbrück Center for Molecular Medicine, Berlin, Germany.|Molecular Cell Biology and Gene Therapy Group, Institute of Biology, Humboldt University, Berlin, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Uckert
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX.
authors.forename: Donna
authors.initials: D
authors.lastname: Palmer
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX.
authors.forename: Philip
authors.initials: P
authors.lastname: Ng
authors.affiliation: Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA.
authors.forename: Kevin G
authors.initials: KG
authors.lastname: Haworth
authors.affiliation: Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA.
authors.forename: Hans-Peter
authors.initials: HP
authors.lastname: Kiem
authors.affiliation: Institute for Virology and Microbiology, University Witten/Herdecke, Witten, Germany.
authors.forename: Anja
authors.initials: A
authors.lastname: Ehrhardt
authors.affiliation: Division of Hematology, Department of Medicine, and.
authors.forename: Thalia
authors.initials: T
authors.lastname: Papayannopoulou
authors.affiliation: Division of Medical Genetics, Department of Medicine, University of Washington, Seattle, WA.|Department of Pathology, University of Washington, Seattle, WA.
authors.forename: André
authors.initials: A
authors.lastname: Lieber
mesh_terms: D000256:Adenoviridae; D000818:Animals; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage; D019650:Hematopoietic Stem Cell Mobilization / Q000379:methods*; D006412:Hematopoietic Stem Cells; D064593:Heterografts; D006801:Humans; D007275:Injections, Intravenous; D051916:Membrane Cofactor Protein / Q000096:biosynthesis*; D051379:Mice; D008810:Mice, Inbred C57BL; D014161:Transduction, Genetic / Q000379:methods*
chemical_list: D051916:Membrane Cofactor Protein