title: Overcoming reprogramming resistance of Fanconi anemia cells.
abstract: Fanconi anemia (FA) is a recessive syndrome characterized by progressive fatal BM failure and chromosomal instability. FA cells have inactivating mutations in a signaling pathway that is critical for maintaining genomic integrity and protecting cells from the DNA damage caused by cross-linking agents. Transgenic expression of the implicated genes corrects the phenotype of hematopoietic cells, but previous attempts at gene therapy have failed largely because of inadequate numbers of hematopoietic stem cells available for gene correction. Induced pluripotent stem cells (iPSCs) constitute an alternate source of autologous cells that are amenable to ex vivo expansion, genetic correction, and molecular characterization. In the present study, we demonstrate that reprogramming leads to activation of the FA pathway, increased DNA double-strand breaks, and senescence. We also demonstrate that defects in the FA DNA-repair pathway decrease the reprogramming efficiency of murine and human primary cells. FA pathway complementation reduces senescence and restores the reprogramming efficiency of somatic FA cells to normal levels. Disease-specific iPSCs derived in this fashion maintain a normal karyotype and are capable of hematopoietic differentiation. These data define the role of the FA pathway in reprogramming and provide a strategy for future translational applications of patient-specific FA iPSCs.
authors.affiliation: Division of Pediatric Hematology/Oncology, Children's Hospital Boston, and Dana-Farber Cancer Institute, Harvard Stem Cell Institute, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Lars U W
authors.initials: LU
authors.lastname: Müller
authors.forename: Michael D
authors.initials: MD
authors.lastname: Milsom
authors.forename: Chad E
authors.initials: CE
authors.lastname: Harris
authors.forename: Rutesh
authors.initials: R
authors.lastname: Vyas
authors.forename: Kristina M
authors.initials: KM
authors.lastname: Brumme
authors.forename: Kalindi
authors.initials: K
authors.lastname: Parmar
authors.forename: Lisa A
authors.initials: LA
authors.lastname: Moreau
authors.forename: Axel
authors.initials: A
authors.lastname: Schambach
authors.forename: In-Hyun
authors.initials: IH
authors.lastname: Park
authors.forename: Wendy B
authors.initials: WB
authors.lastname: London
authors.forename: Kelly
authors.initials: K
authors.lastname: Strait
authors.forename: Thorsten
authors.initials: T
authors.lastname: Schlaeger
authors.forename: Alexander L
authors.initials: AL
authors.lastname: Devine
authors.forename: Elke
authors.initials: E
authors.lastname: Grassman
authors.forename: Alan
authors.initials: A
authors.lastname: D'Andrea
authors.forename: George Q
authors.initials: GQ
authors.lastname: Daley
authors.forename: David A
authors.initials: DA
authors.lastname: Williams
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004249:DNA Damage; D005199:Fanconi Anemia / Q000235:genetics* / Q000378:metabolism / Q000628:therapy; D052217:Fanconi Anemia Complementation Group A Protein / Q000235:genetics; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D017353:Gene Deletion; D015316:Genetic Therapy / Q000379:methods*; D006410:Hematopoiesis; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D059785:Karyotype; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D015398:Signal Transduction
chemical_list: C120277:Fanca protein, mouse; D052217:Fanconi Anemia Complementation Group A Protein