title: A simplified approach to improve the efficiency and safety of ex vivo hematopoietic gene therapy in fanconi anemia patients.
abstract: Fanconi anemia (FA) is an inherited DNA repair disorder characterized by genetic instability of cells lacking a functional FA/BRCA pathway. Previous studies have shown that in vitro stimulation of bone marrow cells (BMCs) from FA mice promotes apoptosis, reduces the reconstitution ability of the stem cells, and induces myelodysplasia and myeloid leukemia upon reinfusion of the cells. This suggests the convenience of adapting standard protocols of gene therapy to FA. Here we show that the reserve of BM progenitors in FA patients is generally below 20% of normal values. Because this reduced reserve could activate the cycling of BM progenitors, we developed a simplified protocol to transduce BMCs from FA patients with gammaretroviral vectors. We demonstrate that a short in vitro manipulation (12-24 hr) of fresh mononuclear BMCs is sufficient to transduce 42% of hematopoietic progenitors from FA-A patients, in the absence of in vitro prestimulation. When FANCA-expressing vectors were used, this simple procedure reversed the phenotype of the BM progenitors from these patients. We propose that our approach will be more efficient and safer compared with standard gene therapy protocols for FA.
authors.affiliation: Spanish Fanconi Anemia Research Network, Madrid 28040, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Jacome
authors.forename: S
authors.initials: S
authors.lastname: Navarro
authors.forename: J A
authors.initials: JA
authors.lastname: Casado
authors.forename: P
authors.initials: P
authors.lastname: Rio
authors.forename: L
authors.initials: L
authors.lastname: Madero
authors.forename: J
authors.initials: J
authors.lastname: Estella
authors.forename: J
authors.initials: J
authors.lastname: Sevilla
authors.forename: I
authors.initials: I
authors.lastname: Badell
authors.forename: J J
authors.initials: JJ
authors.lastname: Ortega
authors.forename: T
authors.initials: T
authors.lastname: Olivé
authors.forename: H
authors.initials: H
authors.lastname: Hanenberg
authors.forename: J C
authors.initials: JC
authors.lastname: Segovia
authors.forename: J A
authors.initials: JA
authors.lastname: Bueren
mesh_terms: D001854:Bone Marrow Cells / Q000502:physiology; D002478:Cells, Cultured; D005199:Fanconi Anemia / Q000097:blood / Q000628:therapy*; D017988:Gammaretrovirus / Q000235:genetics; D015316:Genetic Therapy / Q000009:adverse effects / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000502:physiology*; D006801:Humans; D012016:Reference Values; D014161:Transduction, Genetic / Q000379:methods*