Open AccessTargeted gene therapy and cell reprogramming in F anconi anemia
Author(s) -
Rio Paula,
Baños Rocio,
Lombardo Angelo,
QuintanaBustamante Oscar,
Alvarez Lara,
Garate Zita,
Genovese Pietro,
Almarza Elena,
Valeri Antonio,
Díez Begoña,
Navarro Susana,
Torres Yaima,
Trujillo Juan P,
Murillas Rodolfo,
Segovia Jose C,
Samper Enrique,
Surralles Jordi,
Gregory Philip D,
Holmes Michael C,
Naldini Luigi,
Bueren Juan A
Publication year - 2014
Publication title -
embo molecular medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.923
H-Index - 107
eISSN - 1757-4684
pISSN - 1757-4676
DOI - 10.15252/emmm.201303374
Subject(s) - fanca , reprogramming , biology , zinc finger nuclease , fanconi anemia , genetic enhancement , hematopoietic stem cell , zinc finger , haematopoiesis , gene , genetics , cancer research , microbiology and biotechnology , stem cell , dna repair , transcription factor
Gene targeting is progressively becoming a realistic therapeutic alternative in clinics. It is unknown, however, whether this technology will be suitable for the treatment of DNA repair deficiency syndromes such as F anconi anemia ( FA ), with defects in homology‐directed DNA repair. In this study, we used zinc finger nucleases and integrase‐defective lentiviral vectors to demonstrate for the first time that FANCA can be efficiently and specifically targeted into the AAVS1 safe harbor locus in fibroblasts from FA ‐ A patients. Strikingly, up to 40% of FA fibroblasts showed gene targeting 42 days after gene editing. Given the low number of hematopoietic precursors in the bone marrow of FA patients, gene‐edited FA fibroblasts were then reprogrammed and re‐differentiated toward the hematopoietic lineage. Analyses of gene‐edited FA ‐ iPSC s confirmed the specific integration of FANCA in the AAVS1 locus in all tested clones. Moreover, the hematopoietic differentiation of these iPSC s efficiently generated disease‐free hematopoietic progenitors. Taken together, our results demonstrate for the first time the feasibility of correcting the phenotype of a DNA repair deficiency syndrome using gene‐targeting and cell reprogramming strategies.