title: Phenotypic correction of Fanconi anemia cells in the murine bone marrow after carrier cell mediated delivery of lentiviral vector.
abstract: Fanconi anemia (FA) is an autosomal-recessive disorder associated with hematopoietic failure and it is a candidate for hematopoietic stem cell (HSC)-directed gene therapy. However, the characteristically reduced HSC numbers found in FA patients, their ineffective mobilization from the marrow, and re-oxygenation damage during ex vivo manipulation have precluded clinical success using conventional in vitro approaches. We previously demonstrated that lentiviral vector (LV) particles reversibly attach to the cell surface where they gain protection from serum complement neutralization. We reasoned that cellular delivery of LV to the bone marrow niche could avoid detrimental losses during FA HSC mobilization and in vitro modification. Here, we demonstrate that a VSV-G pseudotyped lentivector, carrying the FANCC transgene, can be transmitted from carrier to bystander cells. In cell culture and transplantation models of FA, we further demonstrate that LV carrier cells migrate along SDF-1α gradients and transfer vector particles that stably integrate and phenotypically correct the characteristic DNA alkylator sensitivity in murine and human FA-deficient target bystander cells. Altogether, we demonstrate that cellular homing mechanisms can be harnessed for the functional phenotype correction in murine FA hematopoietic cells.
authors.affiliation: Department of Pediatrics, Oregon Health & Science University, Portland, OR, 97239, USA. chakkara@ohsu.edu.|Papé Family Pediatric Research Institute, Oregon Health & Science University, L321, Portland, OR, 97239, USA. chakkara@ohsu.edu.
authors.forename: Santhosh
authors.initials: S
authors.lastname: Chakkaramakkil Verghese
authors.affiliation: Department of Pediatrics, Oregon Health & Science University, Portland, OR, 97239, USA.|Papé Family Pediatric Research Institute, Oregon Health & Science University, L321, Portland, OR, 97239, USA.|Present address: Molecular, Cellular, Developmental Biology and Genetics, University of Minnesota, Minneapolis, MN, 55455, USA.
authors.forename: Natalya A
authors.initials: NA
authors.lastname: Goloviznina
authors.affiliation: Department of Pediatrics, Oregon Health & Science University, Portland, OR, 97239, USA.|Papé Family Pediatric Research Institute, Oregon Health & Science University, L321, Portland, OR, 97239, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Kurre
mesh_terms: D000818:Animals; D001853:Bone Marrow / Q000502:physiology*; D002460:Cell Line; D005199:Fanconi Anemia / Q000235:genetics* / Q000473:pathology / Q000628:therapy*; D052218:Fanconi Anemia Complementation Group C Protein / Q000235:genetics; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D057809:HEK293 Cells; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D010641:Phenotype; D019076:Transgenes / Q000235:genetics
chemical_list: D052218:Fanconi Anemia Complementation Group C Protein
keywords: FA gene therapy; Fanconi Anemia; Gene Therapy; Hematopoietic stem cells; In situ gene delivery; Lentiviral vector