A single epidermal stem cell strategy for safe ex vivo gene therapy
Abstract There is a widespread agreement from patient and professional organisations alike that the safety of stem cell therapeutics is of paramount importance, particularly for ex vivo autologous gene therapy. Yet current technology makes it difficult to thoroughly evaluate the behaviour of genetic...
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| Format: | Article |
| Language: | English |
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Springer Nature
2015-02-01
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| Series: | EMBO Molecular Medicine |
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| Online Access: | https://doi.org/10.15252/emmm.201404353 |
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| author | Stéphanie Droz‐Georget Lathion Ariane Rochat Graham Knott Alessandra Recchia Danielle Martinet Sara Benmohammed Nicolas Grasset Andrea Zaffalon Nathalie Besuchet Schmutz Emmanuelle Savioz‐Dayer Jacques Samuel Beckmann Jacques Rougemont Fulvio Mavilio Yann Barrandon |
| author_facet | Stéphanie Droz‐Georget Lathion Ariane Rochat Graham Knott Alessandra Recchia Danielle Martinet Sara Benmohammed Nicolas Grasset Andrea Zaffalon Nathalie Besuchet Schmutz Emmanuelle Savioz‐Dayer Jacques Samuel Beckmann Jacques Rougemont Fulvio Mavilio Yann Barrandon |
| author_sort | Stéphanie Droz‐Georget Lathion |
| collection | DOAJ |
| description | Abstract There is a widespread agreement from patient and professional organisations alike that the safety of stem cell therapeutics is of paramount importance, particularly for ex vivo autologous gene therapy. Yet current technology makes it difficult to thoroughly evaluate the behaviour of genetically corrected stem cells before they are transplanted. To address this, we have developed a strategy that permits transplantation of a clonal population of genetically corrected autologous stem cells that meet stringent selection criteria and the principle of precaution. As a proof of concept, we have stably transduced epidermal stem cells (holoclones) obtained from a patient suffering from recessive dystrophic epidermolysis bullosa. Holoclones were infected with self‐inactivating retroviruses bearing a COL7A1 cDNA and cloned before the progeny of individual stem cells were characterised using a number of criteria. Clonal analysis revealed a great deal of heterogeneity among transduced stem cells in their capacity to produce functional type VII collagen (COLVII). Selected transduced stem cells transplanted onto immunodeficient mice regenerated a non‐blistering epidermis for months and produced a functional COLVII. Safety was assessed by determining the sites of proviral integration, rearrangements and hit genes and by whole‐genome sequencing. The progeny of the selected stem cells also had a diploid karyotype, was not tumorigenic and did not disseminate after long‐term transplantation onto immunodeficient mice. In conclusion, a clonal strategy is a powerful and efficient means of by‐passing the heterogeneity of a transduced stem cell population. It guarantees a safe and homogenous medicinal product, fulfilling the principle of precaution and the requirements of regulatory affairs. Furthermore, a clonal strategy makes it possible to envision exciting gene‐editing technologies like zinc finger nucleases, TALENs and homologous recombination for next‐generation gene therapy. |
| format | Article |
| id | doaj-art-5072a14a0e0b43f2af7cd65c69e46534 |
| institution | Kabale University |
| issn | 1757-4676 1757-4684 |
| language | English |
| publishDate | 2015-02-01 |
| publisher | Springer Nature |
| record_format | Article |
| series | EMBO Molecular Medicine |
| spelling | doaj-art-5072a14a0e0b43f2af7cd65c69e465342025-08-24T11:44:32ZengSpringer NatureEMBO Molecular Medicine1757-46761757-46842015-02-017438039310.15252/emmm.201404353A single epidermal stem cell strategy for safe ex vivo gene therapyStéphanie Droz‐Georget Lathion0Ariane Rochat1Graham Knott2Alessandra Recchia3Danielle Martinet4Sara Benmohammed5Nicolas Grasset6Andrea Zaffalon7Nathalie Besuchet Schmutz8Emmanuelle Savioz‐Dayer9Jacques Samuel Beckmann10Jacques Rougemont11Fulvio Mavilio12Yann Barrandon13Department of Experimental Surgery, Lausanne University Hospital (CHUV)Department of Experimental Surgery, Lausanne University Hospital (CHUV)Interdisciplinary Center for Electron Microscopy, Faculty of Life Sciences EPFLDepartment of Life Sciences, University of Modena and Reggio EmiliaService de Génétique Médicale, Lausanne University Hospital (CHUV)Department of Medical Genetics, Université de LausanneDepartment of Experimental Surgery, Lausanne University Hospital (CHUV)Department of Experimental Surgery, Lausanne University Hospital (CHUV)Service de Génétique Médicale, Lausanne University Hospital (CHUV)Department of Experimental Surgery, Lausanne University Hospital (CHUV)Service de Génétique Médicale, Lausanne University Hospital (CHUV)Bioinformatics and Biostatistics Core Facility, Faculty of Life Sciences EPFLDepartment of Life Sciences, University of Modena and Reggio EmiliaDepartment of Experimental Surgery, Lausanne University Hospital (CHUV)Abstract There is a widespread agreement from patient and professional organisations alike that the safety of stem cell therapeutics is of paramount importance, particularly for ex vivo autologous gene therapy. Yet current technology makes it difficult to thoroughly evaluate the behaviour of genetically corrected stem cells before they are transplanted. To address this, we have developed a strategy that permits transplantation of a clonal population of genetically corrected autologous stem cells that meet stringent selection criteria and the principle of precaution. As a proof of concept, we have stably transduced epidermal stem cells (holoclones) obtained from a patient suffering from recessive dystrophic epidermolysis bullosa. Holoclones were infected with self‐inactivating retroviruses bearing a COL7A1 cDNA and cloned before the progeny of individual stem cells were characterised using a number of criteria. Clonal analysis revealed a great deal of heterogeneity among transduced stem cells in their capacity to produce functional type VII collagen (COLVII). Selected transduced stem cells transplanted onto immunodeficient mice regenerated a non‐blistering epidermis for months and produced a functional COLVII. Safety was assessed by determining the sites of proviral integration, rearrangements and hit genes and by whole‐genome sequencing. The progeny of the selected stem cells also had a diploid karyotype, was not tumorigenic and did not disseminate after long‐term transplantation onto immunodeficient mice. In conclusion, a clonal strategy is a powerful and efficient means of by‐passing the heterogeneity of a transduced stem cell population. It guarantees a safe and homogenous medicinal product, fulfilling the principle of precaution and the requirements of regulatory affairs. Furthermore, a clonal strategy makes it possible to envision exciting gene‐editing technologies like zinc finger nucleases, TALENs and homologous recombination for next‐generation gene therapy.https://doi.org/10.15252/emmm.201404353cell therapyregulatory affairsstem cellswound healing |
| spellingShingle | Stéphanie Droz‐Georget Lathion Ariane Rochat Graham Knott Alessandra Recchia Danielle Martinet Sara Benmohammed Nicolas Grasset Andrea Zaffalon Nathalie Besuchet Schmutz Emmanuelle Savioz‐Dayer Jacques Samuel Beckmann Jacques Rougemont Fulvio Mavilio Yann Barrandon A single epidermal stem cell strategy for safe ex vivo gene therapy EMBO Molecular Medicine cell therapy regulatory affairs stem cells wound healing |
| title | A single epidermal stem cell strategy for safe ex vivo gene therapy |
| title_full | A single epidermal stem cell strategy for safe ex vivo gene therapy |
| title_fullStr | A single epidermal stem cell strategy for safe ex vivo gene therapy |
| title_full_unstemmed | A single epidermal stem cell strategy for safe ex vivo gene therapy |
| title_short | A single epidermal stem cell strategy for safe ex vivo gene therapy |
| title_sort | single epidermal stem cell strategy for safe ex vivo gene therapy |
| topic | cell therapy regulatory affairs stem cells wound healing |
| url | https://doi.org/10.15252/emmm.201404353 |
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