Rescue of multi-lineage hematopoiesis during HIV-1 infection by human c-mpl gene transfer and reconstitution of CD34+ progenitor cells in vivo.
Cytopenias arising from hematopoietic abnormalities are a severe common complication contributing to early mortality in HIV/AIDS patients. The proto-oncogene c-mpl, identified as the thrombopoietin receptor is involved in multilineage differentiation of CD34+ hematopoietic progenitor cells. We have introduced the c-mpl gene into CD34+ cells via transduction of the lentivirus p156RRLsinPPTmPGK-CMPL-PRE. The lentiviral construct expresses c-mpl on approximately 90% of purified CD34+ cells. These transduced cells have then been reconstituted into human fetal thymus/liver implants in severe combined immunodeficient mice (SCID-hu Thy/Liv). The c-mpl expression on transduced CD34+ cells is not susceptible to downregulation due to the effects of HIV-1 infection. Reconstituted CD34+ cells transduced with control lentivirus, p156RRLsinPPTmPGK-EGFP-PRE, express EGFP at > 90%. Reconstituted c-mpl expressing SCID-hu implants show almost maximum rescue (approximately 90%) of myelopoiesis, erythropoiesis and megakaryopoiesis, during HIV-1 infection in vivo, at 6 weeks post-infection. We also show that the differentiated multi-lineage progeny colonies and thymocytes in mice reconstituted with the c-mpl transduced CD34+ cells, carry the HLA Class I loci phenotypes of these donor cells, in the implants of the recipient SCID-hu animals. We propose a gene therapeutic strategy, with c-mpl as the major genetic component, to address the morbidity and mortality resulting from cytopenias in HIV infected patients.
Zhang
Menghua
M
Laboratory of Stem Cell Biology, Torrey Pines Institute for Molecular Studies, 3550 General Atomics Court, San Diego, California 92121, USA.
Poh
Tuang Yeow
TY
Louache
Fawzia
F
Sundell
I Birgitta
IB
Yuan
Jinyun
J
Evans
Stella
S
Koka
Prasad S
PS
D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D019070:Cell Lineage; D002478:Cells, Cultured; D004195:Disease Models, Animal; D053686:Fetal Stem Cells / Q000378:metabolism / Q000637:transplantation*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D015658:HIV Infections / Q000097:blood / Q000235:genetics / Q000628:therapy* / Q000821:virology; D015497:HIV-1 / Q000472:pathogenicity*; D006402:Hematologic Diseases / Q000097:blood / Q000235:genetics / Q000628:therapy* / Q000821:virology; D006410:Hematopoiesis; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000378:metabolism; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008099:Liver / Q000196:embryology / Q000601:surgery; D016031:Liver Transplantation; D051379:Mice; D016513:Mice, SCID; D000090063:Proto-Oncogene Mas; D034622:RNA Interference; D053628:Receptors, Thrombopoietin / Q000096:biosynthesis* / Q000235:genetics; D013950:Thymus Gland / Q000196:embryology / Q000637:transplantation; D013997:Time Factors; D014161:Transduction, Genetic
D018952:Antigens, CD34; C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D053628:Receptors, Thrombopoietin; C082218:MPL protein, human