title: Rapamycin relieves lentiviral vector transduction resistance in human and mouse hematopoietic stem cells.
abstract: Transplantation of genetically modified hematopoietic stem cells (HSCs) is a promising therapeutic strategy for genetic diseases, HIV, and cancer. However, a barrier for clinical HSC gene therapy is the limited efficiency of gene delivery via lentiviral vectors (LVs) into HSCs. We show here that rapamycin, an allosteric inhibitor of the mammalian target of rapamycin complexes, facilitates highly efficient lentiviral transduction of mouse and human HSCs and dramatically enhances marking frequency in long-term engrafting cells in mice. Mechanistically, rapamycin enhanced postbinding endocytic events, leading to increased levels of LV cytoplasmic entry, reverse transcription, and genomic integration. Despite increasing LV copy number, rapamycin did not significantly alter LV integration site profile or chromosomal distribution in mouse HSCs. Rapamycin also enhanced in situ transduction of mouse HSCs via direct intraosseous infusion. Collectively, rapamycin strongly augments LV transduction of HSCs in vitro and in vivo and may prove useful for therapeutic gene delivery.
authors.affiliation: Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA;
authors.forename: Cathy X
authors.initials: CX
authors.lastname: Wang
authors.affiliation: Seattle Children's Research Institute, Seattle, WA;
authors.forename: Blythe D
authors.initials: BD
authors.lastname: Sather
authors.affiliation: Seattle Children's Research Institute, Seattle, WA;
authors.forename: Xuefeng
authors.initials: X
authors.lastname: Wang
authors.affiliation: Fred Hutchinson Cancer Research Center, Seattle, WA; and.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Adair
authors.affiliation: Seattle Children's Research Institute, Seattle, WA;
authors.forename: Iram
authors.initials: I
authors.lastname: Khan
authors.affiliation: Seattle Children's Research Institute, Seattle, WA;
authors.forename: Swati
authors.initials: S
authors.lastname: Singh
authors.affiliation: Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA;
authors.forename: Shanshan
authors.initials: S
authors.lastname: Lang
authors.affiliation: Fred Hutchinson Cancer Research Center, Seattle, WA; and.
authors.forename: Amie
authors.initials: A
authors.lastname: Adams
authors.affiliation: Seattle Children's Research Institute, Seattle, WA;
authors.forename: Gabrielle
authors.initials: G
authors.lastname: Curinga
authors.affiliation: Fred Hutchinson Cancer Research Center, Seattle, WA; and.
authors.forename: Hans-Peter
authors.initials: HP
authors.lastname: Kiem
authors.affiliation: Seattle Children's Research Institute, Seattle, WA;
authors.forename: Carol H
authors.initials: CH
authors.lastname: Miao
authors.affiliation: Seattle Children's Research Institute, Seattle, WA; Department of Pediatrics and Immunology, University of Washington School of Medicine, Seattle, WA.
authors.forename: David J
authors.initials: DJ
authors.lastname: Rawlings
authors.affiliation: Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA;
authors.forename: Bruce E
authors.initials: BE
authors.lastname: Torbett
mesh_terms: D000818:Animals; D005822:Genetic Vectors / Q000187:drug effects; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000187:drug effects* / Q000378:metabolism* / Q000821:virology; D064593:Heterografts; D006801:Humans; D016086:Lentivirus / Q000187:drug effects* / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D020123:Sirolimus / Q000494:pharmacology*; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D014161:Transduction, Genetic / Q000379:methods*; D053586:Virus Internalization / Q000187:drug effects
chemical_list: D058570:TOR Serine-Threonine Kinases; D020123:Sirolimus