title: Engraftment and lineage potential of adult hematopoietic stem and progenitor cells is compromised following short-term culture in the presence of an aryl hydrocarbon receptor antagonist.
abstract: Hematopoietic stem cell gene therapy for HIV/AIDS is a promising alternative to lifelong antiretroviral therapy. One of the limitations of this approach is the number and quality of stem cells available for transplant following in vitro manipulations associated with stem cell isolation and genetic modification. The development of methods to increase the number of autologous, gene-modified stem cells available for transplantation would overcome this barrier. Hematopoietic stem and progenitor cells (HSPC) from adult growth factor-mobilized peripheral blood were cultured in the presence of an aryl hydrocarbon receptor antagonist (AhRA) previously shown to expand HSPC from umbilical cord blood. Qualitative and quantitative assessment of the hematopoietic potential of minimally cultured (MC-HSPC) or expanded HSPC (Exp-HSPC) was performed using an immunodeficient mouse model of transplantation. Our results demonstrate robust, multilineage engraftment of both MC-HSPC and Exp-HSPC although estimates of expansion based on stem cell phenotype were not supported by a corresponding increase in in vivo engrafting units. Bone marrow of animals transplanted with either MC-HSPC or Exp-HSPC contained secondary engrafting cells verifying the presence of primitive stem cells in both populations. However, the frequency of in vivo engrafting units among the more primitive CD34+/CD90+ HSPC population was significantly lower in Exp-HSPC compared with MC-HSPC. Exp-HSPC also produced fewer lymphoid progeny and more myeloid progeny than MC-HSPC. These results reveal that in vitro culture of adult HSPC in AhRA maintains but does not increase the number of in vivo engrafting cells and that HSPC expanded in vitro contain defects in lymphopoiesis as assessed in this model system. Further investigation is required before implementation of this approach in the clinical setting.
authors.affiliation: 1 Laboratory for Cellular Medicine, Beckman Research Institute , City of Hope, CA 91010.
authors.forename: Angel
authors.initials: A
authors.lastname: Gu
authors.forename: Monica
authors.initials: M
authors.lastname: Torres-Coronado
authors.forename: Chy-Anh
authors.initials: CA
authors.lastname: Tran
authors.forename: Hieu
authors.initials: H
authors.lastname: Vu
authors.forename: Elizabeth W
authors.initials: EW
authors.lastname: Epps
authors.forename: Janet
authors.initials: J
authors.lastname: Chung
authors.forename: Nancy
authors.initials: N
authors.lastname: Gonzalez
authors.forename: Suzette
authors.initials: S
authors.lastname: Blanchard
authors.forename: David L
authors.initials: DL
authors.lastname: DiGiusto
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D001391:Azo Compounds / Q000494:pharmacology; D019070:Cell Lineage; D002478:Cells, Cultured; D005312:Fetal Blood / Q000166:cytology; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D017493:Leukocyte Common Antigens / Q000378:metabolism; D051379:Mice; D016688:Mice, Inbred NOD; D023421:Models, Animal; D010641:Phenotype; D011720:Pyrazoles / Q000494:pharmacology; D018336:Receptors, Aryl Hydrocarbon / Q000037:antagonists & inhibitors* / Q000378:metabolism; D018800:Thy-1 Antigens / Q000378:metabolism; D014183:Transplantation, Heterologous
chemical_list: C511621:2-methyl-2H-pyrazole-3-carboxylic acid (2-methyl-4-o-tolylazophenyl)amide; D018952:Antigens, CD34; D001391:Azo Compounds; D011720:Pyrazoles; D018336:Receptors, Aryl Hydrocarbon; D018800:Thy-1 Antigens; D017493:Leukocyte Common Antigens