Improved transduction of human sheep repopulating cells by retrovirus vectors pseudotyped with feline leukemia virus type C or RD114 envelopes.
Gene therapy for hematopoietic diseases has been hampered by the low frequency of transduction of human hematopoietic stem cells (HSCs) with retroviral vectors pseudotyped with amphotropic envelopes. We hypothesized that transduction could be increased by the use of retroviral vectors pseudotyped with envelopes that recognize more abundant cellular receptors. The levels of mRNA encoding the receptors of the feline retroviruses, RD114 and feline leukemia virus type C (FeLV-C), were significantly higher than the level of gibbon ape leukemia virus (GaLV) receptor mRNA in cells enriched for human HSCs (Lin- CD34+ CD38-). We cotransduced human peripheral blood CD34+ cells with equivalent numbers of FeLV-C and GALV or RD114 and GALV-pseudotyped retroviruses for injection into fetal sheep. Analysis of DNA from peripheral blood and bone marrow from recipient sheep demonstrated that FeLV-C- or RD114-pseudotyped vectors were present at significantly higher levels than GALV-pseudotyped vectors. Analysis of individual myeloid colonies demonstrated that retrovirus vectors with FeLV-C and RD114 pseudotypes were present at 1.5 to 1.6 copies per cell and were preferentially integrated near known genes We conclude that the more efficient transduction of human HSCs with either FeLV-C- or RD114-pseudotyped retroviral particles may improve gene transfer in human clinical trials.
Lucas
M Lee
ML
Genetics and Molecular Biology Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Seidel
Nancy E
NE
Porada
Christopher D
CD
Quigley
John G
JG
Anderson
Stacie M
SM
Malech
Harry L
HL
Abkowitz
Janis L
JL
Zanjani
Esmail D
ED
Bodine
David M
DM
D026962:Amino Acid Transport System ASC / Q000235:genetics*; D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D001854:Bone Marrow Cells / Q000502:physiology; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006367:HeLa Cells; D006402:Hematologic Diseases / Q000628:therapy; D006801:Humans; D020014:K562 Cells; D007763:Lac Operon; D026901:Membrane Transport Proteins / Q000235:genetics*; D015778:Minor Histocompatibility Antigens; D012333:RNA, Messenger / Q000032:analysis; D011991:Receptors, Virus / Q000235:genetics*; D012190:Retroviridae / Q000235:genetics*; D012756:Sheep; D014161:Transduction, Genetic / Q000379:methods*; D016662:Virus Integration
D026962:Amino Acid Transport System ASC; D018952:Antigens, CD34; C492690:FLVCR1 protein, human; D026901:Membrane Transport Proteins; D015778:Minor Histocompatibility Antigens; D012333:RNA, Messenger; D011991:Receptors, Virus; C420506:SLC1A5 protein, human