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.
Laboratory of Stem Cell Biology, Torrey Pines Institute for Molecular Studies, 3550 General Atomics Court, San Diego, California 92121, USA.
Menghua
M
Zhang
Tuang Yeow
TY
Poh
Fawzia
F
Louache
I Birgitta
IB
Sundell
Jinyun
J
Yuan
Stella
S
Evans
Prasad S
PS
Koka
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