CXCR4-transgene expression significantly improves marrow engraftment of cultured hematopoietic stem cells.
Hematopoietic stem cells (HSCs) lose marrow reconstitution potential during ex vivo culture. HSC migration to stromal cell-derived factor (SDF)-1 (CXCL12) correlates with CXC chemokine receptor 4 (CXCR4) expression and marrow engraftment. We demonstrate that mobilized human CD34+ peripheral blood stem cells (CD34+ PBSCs) lose CXCR4 expression during prolonged culture. We transduced CD34+ PBSCs with retrovirus vector encoding human CXCR4 and achieved 18-fold more CXCR4 expression in over 87% of CD34+ cells. CXCR4-transduced cells yielded increased calcium flux and up to a 10-fold increase in migration to SDF-1. Six-day cultured CXCR4-transduced cells demonstrated significant engraftment in nonobese diabetic/severe combined immunodeficient mice under conditions in which control transduced cells resulted in low or no engraftment. We conclude that transduction-mediated overexpression of CXCR4 significantly improves marrow engraftment of cultured PBSCs.
Brenner
Sebastian
S
Laboratory of Host Defenses, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA. Sebastian.Brenner@uniklinikum-dresden.de
Whiting-Theobald
Narda
N
Kawai
Toshinao
T
Linton
Gilda F
GF
Rudikoff
Andrew G
AG
Choi
Uimook
U
Ryser
Martin F
MF
Murphy
Philip M
PM
Sechler
Joan M G
JM
Malech
Harry L
HL
D000818:Animals; D018952:Antigens, CD34 / Q000096:biosynthesis; D001854:Bone Marrow Cells / Q000166:cytology*; D002118:Calcium / Q000378:metabolism; D002465:Cell Movement; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D006085:Graft Survival; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D016513:Mice, SCID; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000378:metabolism; D019718:Receptors, CXCR4 / Q000096:biosynthesis* / Q000235:genetics*; D015398:Signal Transduction; D013997:Time Factors; D019076:Transgenes
D018952:Antigens, CD34; D008562:Membrane Glycoproteins; D019718:Receptors, CXCR4; C056558:CYBB protein, human; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases; D002118:Calcium