Overnight transduction with foamyviral vectors restores the long-term repopulating activity of Fancc-/- stem cells.
Fanconi anemia (FA) is a complex genetic disorder characterized by congenital abnormalities, bone marrow failure, and myeloid malignancies. Identification of 13 FA genes has been instrumental to explore gene transfer technologies aimed at correction of autologous FA-deficient stem cells. To date, 3 human FA stem cell gene therapy trials with standard 4-day transduction protocols using gammaretroviral vectors failed to provide clinical benefit. In addition, 2- to 4 day ex vivo manipulation of bone marrow from mice containing a disruption of the homologue of human FANCC (Fancc) results in a time-dependent increase in apoptosis and a risk for malignant transformation of hematopoietic cells. Here, we show that a 14-hour transduction period allows a foamyviral vector construct expressing the human FANCC cDNA to efficiently transduce murine FA stem cells with 1 to 2 proviral integrations per genome. Functionally, the repopulating activity of Fancc(-/-) stem cells from reconstituted mice expressing the recombinant FANCC transgene was comparable with wild-type controls. Collectively, these data provide evidence that short-term transduction of c-kit(+) cells with a foamyviral vector is sufficient for functional correction of a stem cell phenotype in a murine FA model. These data could have implications for future gene therapy trials for FA patients.
Si
Yue
Y
Departments of Microbiology and Immunology, Herman B Wells Center for Pediatric Research, Indiana University School of Medicine,Indianapolis, USA.
Pulliam
Anna C
AC
Linka
Yvonne
Y
Ciccone
Samantha
S
Leurs
Cordula
C
Yuan
Jin
J
Eckermann
Olaf
O
Fruehauf
Stefan
S
Mooney
Sean
S
Hanenberg
Helmut
H
Clapp
D Wade
DW
D000818:Animals; D049109:Cell Proliferation; D002478:Cells, Cultured; D002940:Circadian Rhythm; D005199:Fanconi Anemia / Q000235:genetics / Q000473:pathology / Q000628:therapy* / Q000662:veterinary; D052218:Fanconi Anemia Complementation Group C Protein / Q000235:genetics* / Q000378:metabolism; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods; D006412:Hematopoietic Stem Cells / Q000378:metabolism / Q000473:pathology / Q000502:physiology*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics; D016092:Spumavirus / Q000235:genetics*; D013997:Time Factors; D014161:Transduction, Genetic / Q000379:methods*
C497652:Fancc protein, mouse; D052218:Fanconi Anemia Complementation Group C Protein; D019009:Proto-Oncogene Proteins c-kit