Targeted gene therapy and cell reprogramming in Fanconi anemia
Abstract 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 Fanconi anemia (FA), with defects in homology‐directed DNA repair. In this s...
| Main Authors: | , , , , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Springer Nature
2014-06-01
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| Series: | EMBO Molecular Medicine |
| Subjects: | |
| Online Access: | https://doi.org/10.15252/emmm.201303374 |
| _version_ | 1797280661995782144 |
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| author | Paula Rio Rocio Baños Angelo Lombardo Oscar Quintana‐Bustamante Lara Alvarez Zita Garate Pietro Genovese Elena Almarza Antonio Valeri Begoña Díez Susana Navarro Yaima Torres Juan P Trujillo Rodolfo Murillas Jose C Segovia Enrique Samper Jordi Surralles Philip D Gregory Michael C Holmes Luigi Naldini Juan A Bueren |
| author_facet | Paula Rio Rocio Baños Angelo Lombardo Oscar Quintana‐Bustamante Lara Alvarez Zita Garate Pietro Genovese Elena Almarza Antonio Valeri Begoña Díez Susana Navarro Yaima Torres Juan P Trujillo Rodolfo Murillas Jose C Segovia Enrique Samper Jordi Surralles Philip D Gregory Michael C Holmes Luigi Naldini Juan A Bueren |
| author_sort | Paula Rio |
| collection | DOAJ |
| description | Abstract 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 Fanconi 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‐iPSCs confirmed the specific integration of FANCA in the AAVS1 locus in all tested clones. Moreover, the hematopoietic differentiation of these iPSCs 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. |
| first_indexed | 2024-03-07T16:44:32Z |
| format | Article |
| id | doaj.art-9ea6a100ad2d43d6af6bfa973b399ceb |
| institution | Directory Open Access Journal |
| issn | 1757-4676 1757-4684 |
| language | English |
| last_indexed | 2024-03-07T16:44:32Z |
| publishDate | 2014-06-01 |
| publisher | Springer Nature |
| record_format | Article |
| series | EMBO Molecular Medicine |
| spelling | doaj.art-9ea6a100ad2d43d6af6bfa973b399ceb2024-03-03T07:05:16ZengSpringer NatureEMBO Molecular Medicine1757-46761757-46842014-06-016683584810.15252/emmm.201303374Targeted gene therapy and cell reprogramming in Fanconi anemiaPaula Rio0Rocio Baños1Angelo Lombardo2Oscar Quintana‐Bustamante3Lara Alvarez4Zita Garate5Pietro Genovese6Elena Almarza7Antonio Valeri8Begoña Díez9Susana Navarro10Yaima Torres11Juan P Trujillo12Rodolfo Murillas13Jose C Segovia14Enrique Samper15Jordi Surralles16Philip D Gregory17Michael C Holmes18Luigi Naldini19Juan A Bueren20Division of Hematopoietic Innovative Therapies CIEMAT/CIBERER Madrid SpainDivision of Hematopoietic Innovative Therapies CIEMAT/CIBERER Madrid SpainSan Raffaele Telethon Institute for Gene Therapy San Raffaele Scientific Institute Milan ItalyDivision of Hematopoietic Innovative Therapies CIEMAT/CIBERER Madrid SpainDivision of Hematopoietic Innovative Therapies CIEMAT/CIBERER Madrid SpainDivision of Hematopoietic Innovative Therapies CIEMAT/CIBERER Madrid SpainSan Raffaele Telethon Institute for Gene Therapy San Raffaele Scientific Institute Milan ItalyDivision of Hematopoietic Innovative Therapies CIEMAT/CIBERER Madrid SpainDivision of Hematopoietic Innovative Therapies CIEMAT/CIBERER Madrid SpainDivision of Hematopoietic Innovative Therapies CIEMAT/CIBERER Madrid SpainDivision of Hematopoietic Innovative Therapies CIEMAT/CIBERER Madrid SpainNIMGenetics SL Madrid SpainInstituto de Investigación Sanitaria Fundación Jiménez Díaz (IIS‐FJD, UAM) Madrid SpainDivision of Epithelial Biomedicine CIEMAT/CIBERER Madrid SpainDivision of Hematopoietic Innovative Therapies CIEMAT/CIBERER Madrid SpainNIMGenetics SL Madrid SpainUniversidad Autónoma Barcelona/CIBERER Barcelona SpainSangamo BioSciences Inc. Richmond CA USASangamo BioSciences Inc. Richmond CA USASan Raffaele Telethon Institute for Gene Therapy San Raffaele Scientific Institute Milan ItalyDivision of Hematopoietic Innovative Therapies CIEMAT/CIBERER Madrid SpainAbstract 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 Fanconi 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‐iPSCs confirmed the specific integration of FANCA in the AAVS1 locus in all tested clones. Moreover, the hematopoietic differentiation of these iPSCs 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.https://doi.org/10.15252/emmm.201303374cell reprogrammingFanconi anemiagene‐targetingiPSCszinc finger nucleases |
| spellingShingle | Paula Rio Rocio Baños Angelo Lombardo Oscar Quintana‐Bustamante Lara Alvarez Zita Garate Pietro Genovese Elena Almarza Antonio Valeri Begoña Díez Susana Navarro Yaima Torres Juan P Trujillo Rodolfo Murillas Jose C Segovia Enrique Samper Jordi Surralles Philip D Gregory Michael C Holmes Luigi Naldini Juan A Bueren Targeted gene therapy and cell reprogramming in Fanconi anemia EMBO Molecular Medicine cell reprogramming Fanconi anemia gene‐targeting iPSCs zinc finger nucleases |
| title | Targeted gene therapy and cell reprogramming in Fanconi anemia |
| title_full | Targeted gene therapy and cell reprogramming in Fanconi anemia |
| title_fullStr | Targeted gene therapy and cell reprogramming in Fanconi anemia |
| title_full_unstemmed | Targeted gene therapy and cell reprogramming in Fanconi anemia |
| title_short | Targeted gene therapy and cell reprogramming in Fanconi anemia |
| title_sort | targeted gene therapy and cell reprogramming in fanconi anemia |
| topic | cell reprogramming Fanconi anemia gene‐targeting iPSCs zinc finger nucleases |
| url | https://doi.org/10.15252/emmm.201303374 |
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