Therapeutic gene editing in CD34+ hematopoietic progenitors from Fanconi anemia patients
Abstract Gene targeting constitutes a new step in the development of gene therapy for inherited diseases. Although previous studies have shown the feasibility of editing fibroblasts from Fanconi anemia (FA) patients, here we aimed at conducting therapeutic gene editing in clinically relevant cells,...
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| Format: | Article |
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Springer Nature
2017-09-01
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| Series: | EMBO Molecular Medicine |
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| Online Access: | https://doi.org/10.15252/emmm.201707540 |
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| author | Begoña Diez Pietro Genovese Francisco J Roman‐Rodriguez Lara Alvarez Giulia Schiroli Laura Ugalde Sandra Rodriguez‐Perales Julian Sevilla Cristina Diaz de Heredia Michael C Holmes Angelo Lombardo Luigi Naldini Juan Antonio Bueren Paula Rio |
| author_facet | Begoña Diez Pietro Genovese Francisco J Roman‐Rodriguez Lara Alvarez Giulia Schiroli Laura Ugalde Sandra Rodriguez‐Perales Julian Sevilla Cristina Diaz de Heredia Michael C Holmes Angelo Lombardo Luigi Naldini Juan Antonio Bueren Paula Rio |
| author_sort | Begoña Diez |
| collection | DOAJ |
| description | Abstract Gene targeting constitutes a new step in the development of gene therapy for inherited diseases. Although previous studies have shown the feasibility of editing fibroblasts from Fanconi anemia (FA) patients, here we aimed at conducting therapeutic gene editing in clinically relevant cells, such as hematopoietic stem cells (HSCs). In our first experiments, we showed that zinc finger nuclease (ZFN)‐mediated insertion of a non‐therapeutic EGFP‐reporter donor in the AAVS1 “safe harbor” locus of FA‐A lymphoblastic cell lines (LCLs), indicating that FANCA is not essential for the editing of human cells. When the same approach was conducted with therapeutic FANCA donors, an efficient phenotypic correction of FA‐A LCLs was obtained. Using primary cord blood CD34+ cells from healthy donors, gene targeting was confirmed not only in in vitro cultured cells, but also in hematopoietic precursors responsible for the repopulation of primary and secondary immunodeficient mice. Moreover, when similar experiments were conducted with mobilized peripheral blood CD34+ cells from FA‐A patients, we could demonstrate for the first time that gene targeting in primary hematopoietic precursors from FA patients is feasible and compatible with the phenotypic correction of these clinically relevant cells. |
| format | Article |
| id | doaj-art-7498c7c0c5d64ba6b01dff08dbecd260 |
| institution | DOAJ |
| issn | 1757-4676 1757-4684 |
| language | English |
| publishDate | 2017-09-01 |
| publisher | Springer Nature |
| record_format | Article |
| series | EMBO Molecular Medicine |
| spelling | doaj-art-7498c7c0c5d64ba6b01dff08dbecd2602025-08-20T03:06:00ZengSpringer NatureEMBO Molecular Medicine1757-46761757-46842017-09-019111574158810.15252/emmm.201707540Therapeutic gene editing in CD34+ hematopoietic progenitors from Fanconi anemia patientsBegoña Diez0Pietro Genovese1Francisco J Roman‐Rodriguez2Lara Alvarez3Giulia Schiroli4Laura Ugalde5Sandra Rodriguez‐Perales6Julian Sevilla7Cristina Diaz de Heredia8Michael C Holmes9Angelo Lombardo10Luigi Naldini11Juan Antonio Bueren12Paula Rio13Division of Hematopoietic Innovative Therapies, Centro de Investigaciones Energéticas, Medioambientales y TecnológicasSan Raffaele Telethon Institute for Gene Therapy (SR‐Tiget), IRCCS San Raffaele Scientific InstituteDivision of Hematopoietic Innovative Therapies, Centro de Investigaciones Energéticas, Medioambientales y TecnológicasDivision of Hematopoietic Innovative Therapies, Centro de Investigaciones Energéticas, Medioambientales y TecnológicasSan Raffaele Telethon Institute for Gene Therapy (SR‐Tiget), IRCCS San Raffaele Scientific InstituteDivision of Hematopoietic Innovative Therapies, Centro de Investigaciones Energéticas, Medioambientales y TecnológicasMolecular Cytogenetics Group, Human Cancer Genetics Program, Centro Nacional de Investigaciones Oncológicas (CNIO)Centro de Investigación Biomédica en Red de Enfermedades RarasServicio de Oncología y Hematología Pediátrica, Hospital Universitario Vall d'HebronSangamo Therapeutics, Inc.San Raffaele Telethon Institute for Gene Therapy (SR‐Tiget), IRCCS San Raffaele Scientific InstituteSan Raffaele Telethon Institute for Gene Therapy (SR‐Tiget), IRCCS San Raffaele Scientific InstituteDivision of Hematopoietic Innovative Therapies, Centro de Investigaciones Energéticas, Medioambientales y TecnológicasDivision of Hematopoietic Innovative Therapies, Centro de Investigaciones Energéticas, Medioambientales y TecnológicasAbstract Gene targeting constitutes a new step in the development of gene therapy for inherited diseases. Although previous studies have shown the feasibility of editing fibroblasts from Fanconi anemia (FA) patients, here we aimed at conducting therapeutic gene editing in clinically relevant cells, such as hematopoietic stem cells (HSCs). In our first experiments, we showed that zinc finger nuclease (ZFN)‐mediated insertion of a non‐therapeutic EGFP‐reporter donor in the AAVS1 “safe harbor” locus of FA‐A lymphoblastic cell lines (LCLs), indicating that FANCA is not essential for the editing of human cells. When the same approach was conducted with therapeutic FANCA donors, an efficient phenotypic correction of FA‐A LCLs was obtained. Using primary cord blood CD34+ cells from healthy donors, gene targeting was confirmed not only in in vitro cultured cells, but also in hematopoietic precursors responsible for the repopulation of primary and secondary immunodeficient mice. Moreover, when similar experiments were conducted with mobilized peripheral blood CD34+ cells from FA‐A patients, we could demonstrate for the first time that gene targeting in primary hematopoietic precursors from FA patients is feasible and compatible with the phenotypic correction of these clinically relevant cells.https://doi.org/10.15252/emmm.201707540CD34+ cellsFanconi anemiagene editinghematopoietic stem and progenitor cellszinc finger nucleases |
| spellingShingle | Begoña Diez Pietro Genovese Francisco J Roman‐Rodriguez Lara Alvarez Giulia Schiroli Laura Ugalde Sandra Rodriguez‐Perales Julian Sevilla Cristina Diaz de Heredia Michael C Holmes Angelo Lombardo Luigi Naldini Juan Antonio Bueren Paula Rio Therapeutic gene editing in CD34+ hematopoietic progenitors from Fanconi anemia patients EMBO Molecular Medicine CD34+ cells Fanconi anemia gene editing hematopoietic stem and progenitor cells zinc finger nucleases |
| title | Therapeutic gene editing in CD34+ hematopoietic progenitors from Fanconi anemia patients |
| title_full | Therapeutic gene editing in CD34+ hematopoietic progenitors from Fanconi anemia patients |
| title_fullStr | Therapeutic gene editing in CD34+ hematopoietic progenitors from Fanconi anemia patients |
| title_full_unstemmed | Therapeutic gene editing in CD34+ hematopoietic progenitors from Fanconi anemia patients |
| title_short | Therapeutic gene editing in CD34+ hematopoietic progenitors from Fanconi anemia patients |
| title_sort | therapeutic gene editing in cd34 hematopoietic progenitors from fanconi anemia patients |
| topic | CD34+ cells Fanconi anemia gene editing hematopoietic stem and progenitor cells zinc finger nucleases |
| url | https://doi.org/10.15252/emmm.201707540 |
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