title: Ex vivo culture of Fancc-/- stem/progenitor cells predisposes cells to undergo apoptosis, and surviving stem/progenitor cells display cytogenetic abnormalities and an increased risk of malignancy.
abstract: Current strategies for genetic therapy using Moloney retroviruses require ex vivo manipulation of hematopoietic cells to facilitate stable integration of the transgene. While many studies have evaluated the impact of ex vivo culture on normal murine and human stem/progenitor cells, the cellular consequences of ex vivo manipulation of stem cells with intrinsic defects in genome stability are incompletely understood. Here we show that ex vivo culture of Fancc(-/-) bone marrow cells results in a time-dependent increase in apoptosis of primitive Fancc(-/-) progenitor cells in conditions that promote the proliferation of wild-type stem/progenitor cells. Further, recipients reconstituted with the surviving Fancc(-/-) cells have a high incidence of cytogenetic abnormalities and myeloid malignancies that are associated with an acquired resistance to tumor necrosis factor alpha (TNF-alpha). Collectively, these data indicate that the intrinsic defects in the genomic stability of Fancc(-/-) stem/progenitor cells provide a selective pressure for cells that are resistant to apoptosis and have a propensity for the evolution to clonal hematopoiesis and malignancy. These studies could have implications for the design of genetic therapies for treatment of Fanconi anemia and potentially other genetic diseases with intrinsic defects in genome stability.
authors.affiliation: Cancer Research Institute, 1044 W Walnut Street, Rm 408, Indianapolis, IN 46202-5254, USA.
authors.forename: Xiaxin
authors.initials: X
authors.lastname: Li
authors.forename: Michelle M
authors.initials: MM
authors.lastname: Le Beau
authors.forename: Samantha
authors.initials: S
authors.lastname: Ciccone
authors.forename: Feng-Chun
authors.initials: FC
authors.lastname: Yang
authors.forename: Brian
authors.initials: B
authors.lastname: Freie
authors.forename: Shi
authors.initials: S
authors.lastname: Chen
authors.forename: Jin
authors.initials: J
authors.lastname: Yuan
authors.forename: Ping
authors.initials: P
authors.lastname: Hong
authors.forename: Attilio
authors.initials: A
authors.lastname: Orazi
authors.forename: Laura S
authors.initials: LS
authors.lastname: Haneline
authors.forename: D Wade
authors.initials: DW
authors.lastname: Clapp
mesh_terms: D000818:Animals; D017209:Apoptosis; D001854:Bone Marrow Cells / Q000473:pathology; D018797:Cell Cycle Proteins / Q000235:genetics; D002478:Cells, Cultured; D002869:Chromosome Aberrations; D002999:Clone Cells; D004268:DNA-Binding Proteins / Q000172:deficiency* / Q000235:genetics; D005199:Fanconi Anemia / Q000473:pathology / Q000628:therapy; D052218:Fanconi Anemia Complementation Group C Protein; D051856:Fanconi Anemia Complementation Group Proteins; D019337:Hematologic Neoplasms / Q000209:etiology*; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects; D006412:Hematopoietic Stem Cells / Q000473:pathology*; D051379:Mice; D018345:Mice, Knockout; D009687:Nuclear Proteins / Q000172:deficiency* / Q000235:genetics; D012306:Risk; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; C497652:Fancc protein, mouse; D052218:Fanconi Anemia Complementation Group C Protein; D051856:Fanconi Anemia Complementation Group Proteins; D009687:Nuclear Proteins; D014409:Tumor Necrosis Factor-alpha