Identification of human and mouse hematopoietic stem cell populations expressing high levels of mRNA encoding retrovirus receptors.
One obstacle to retrovirus-mediated gene therapy for human hematopoietic disorders is the low efficiency of gene transfer into pluripotent hematopoietic stem cells (HSC). We have previously shown a direct correlation between retrovirus receptor mRNA levels in mouse HSC and the efficiency with which they are transduced. In the present study, we assayed retrovirus receptor mRNA levels in a variety of mouse and human HSC populations to identify HSC which may be more competent for retrovirus transduction. The highest levels of amphotropic retrovirus receptor (amphoR) mRNA were found in cryopreserved human cord blood HSC. The level of amphoR mRNA in Lin- CD34(+) CD38(-) cells isolated from frozen cord blood was 12-fold higher than the level in fresh cord blood Lin- CD34(+) CD38(-) cells. In mice, the level of amphoR mRNA in HSC from the bone marrow (BM) of mice treated with stem cell factor and granulocyte-colony stimulating factor was 2.8- to 7.8-fold higher than in HSC from the BM of untreated mice. These findings suggest that HSC from frozen cord blood and cytokine-mobilized BM may be superior targets for amphotropic retrovirus transduction compared with HSC from untreated adult BM.
Orlic
D
D
Hematopoiesis Section, Laboratory of Gene Transfer, National Human Genome Research Institute (NHGRI), National Institutes of Health (NIH), Bethesda, MD, USA.
Girard
L J
LJ
Anderson
S M
SM
Pyle
L C
LC
Yoder
M C
MC
Broxmeyer
H E
HE
Bodine
D M
DM
D000818:Animals; D002352:Carrier Proteins / Q000235:genetics; D002469:Cell Separation; D015870:Gene Expression; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000821:virology; D006801:Humans; D008099:Liver / Q000196:embryology; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D011991:Receptors, Virus / Q000235:genetics*; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic; D015017:Yolk Sac / Q000166:cytology
D002352:Carrier Proteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D012333:RNA, Messenger; D012367:RNA, Viral; D011991:Receptors, Virus; C070506:ecotropic murine leukemia virus receptor