Generation of healthy mice from gene-corrected disease-specific induced pluripotent stem cells.

2.50
HDL Handle:
http://hdl.handle.net/10033/209489
Title:
Generation of healthy mice from gene-corrected disease-specific induced pluripotent stem cells.
Authors:
Wu, Guangming; Liu, Na; Rittelmeyer, Ina; Sharma, Amar Deep; Sgodda, Malte; Zaehres, Holm; Bleidissel, Martina; Greber, Boris; Gentile, Luca; Han, Dong Wook; Rudolph, Cornelia; Steinemann, Doris; Schambach, Axel; Ott, Michael; Schöler, Hans R; Cantz, Tobias
Abstract:
Using the murine model of tyrosinemia type 1 (fumarylacetoacetate hydrolase [FAH] deficiency; FAH⁻/⁻ mice) as a paradigm for orphan disorders, such as hereditary metabolic liver diseases, we evaluated fibroblast-derived FAH⁻/⁻-induced pluripotent stem cells (iPS cells) as targets for gene correction in combination with the tetraploid embryo complementation method. First, after characterizing the FAH⁻/⁻ iPS cell lines, we aggregated FAH⁻/⁻-iPS cells with tetraploid embryos and obtained entirely FAH⁻/⁻-iPS cell-derived mice that were viable and exhibited the phenotype of the founding FAH⁻/⁻ mice. Then, we transduced FAH cDNA into the FAH⁻/⁻-iPS cells using a third-generation lentiviral vector to generate gene-corrected iPS cells. We could not detect any chromosomal alterations in these cells by high-resolution array CGH analysis, and after their aggregation with tetraploid embryos, we obtained fully iPS cell-derived healthy mice with an astonishing high efficiency for full-term development of up to 63.3%. The gene correction was validated functionally by the long-term survival and expansion of FAH-positive cells of these mice after withdrawal of the rescuing drug NTBC (2-(2-nitro-4-fluoromethylbenzoyl)-1,3-cyclohexanedione). Furthermore, our results demonstrate that both a liver-specific promoter (transthyretin, TTR)-driven FAH transgene and a strong viral promoter (from spleen focus-forming virus, SFFV)-driven FAH transgene rescued the FAH-deficiency phenotypes in the mice derived from the respective gene-corrected iPS cells. In conclusion, our data demonstrate that a lentiviral gene repair strategy does not abrogate the full pluripotent potential of fibroblast-derived iPS cells, and genetic manipulation of iPS cells in combination with tetraploid embryo aggregation provides a practical and rapid approach to evaluate the efficacy of gene correction of human diseases in mouse models.
Affiliation:
Max-Planck-Institute for Molecular Biomedicine, Münster, Germany.
Citation:
Generation of healthy mice from gene-corrected disease-specific induced pluripotent stem cells. 2011, 9 (7):e1001099 PLoS Biol.
Journal:
PLoS biology
Issue Date:
Jul-2011
URI:
http://hdl.handle.net/10033/209489
DOI:
10.1371/journal.pbio.1001099
PubMed ID:
21765802
Type:
Article
Language:
en
ISSN:
1545-7885
Appears in Collections:
publications of AG Cell and Gene therapy

Full metadata record

DC FieldValueLanguage
dc.contributor.authorWu, Guangmingen
dc.contributor.authorLiu, Naen
dc.contributor.authorRittelmeyer, Inaen
dc.contributor.authorSharma, Amar Deepen
dc.contributor.authorSgodda, Malteen
dc.contributor.authorZaehres, Holmen
dc.contributor.authorBleidissel, Martinaen
dc.contributor.authorGreber, Borisen
dc.contributor.authorGentile, Lucaen
dc.contributor.authorHan, Dong Wooken
dc.contributor.authorRudolph, Corneliaen
dc.contributor.authorSteinemann, Dorisen
dc.contributor.authorSchambach, Axelen
dc.contributor.authorOtt, Michaelen
dc.contributor.authorSchöler, Hans Ren
dc.contributor.authorCantz, Tobiasen
dc.date.accessioned2012-02-06T15:29:28Z-
dc.date.available2012-02-06T15:29:28Z-
dc.date.issued2011-07-
dc.identifier.citationGeneration of healthy mice from gene-corrected disease-specific induced pluripotent stem cells. 2011, 9 (7):e1001099 PLoS Biol.en
dc.identifier.issn1545-7885-
dc.identifier.pmid21765802-
dc.identifier.doi10.1371/journal.pbio.1001099-
dc.identifier.urihttp://hdl.handle.net/10033/209489-
dc.description.abstractUsing the murine model of tyrosinemia type 1 (fumarylacetoacetate hydrolase [FAH] deficiency; FAH⁻/⁻ mice) as a paradigm for orphan disorders, such as hereditary metabolic liver diseases, we evaluated fibroblast-derived FAH⁻/⁻-induced pluripotent stem cells (iPS cells) as targets for gene correction in combination with the tetraploid embryo complementation method. First, after characterizing the FAH⁻/⁻ iPS cell lines, we aggregated FAH⁻/⁻-iPS cells with tetraploid embryos and obtained entirely FAH⁻/⁻-iPS cell-derived mice that were viable and exhibited the phenotype of the founding FAH⁻/⁻ mice. Then, we transduced FAH cDNA into the FAH⁻/⁻-iPS cells using a third-generation lentiviral vector to generate gene-corrected iPS cells. We could not detect any chromosomal alterations in these cells by high-resolution array CGH analysis, and after their aggregation with tetraploid embryos, we obtained fully iPS cell-derived healthy mice with an astonishing high efficiency for full-term development of up to 63.3%. The gene correction was validated functionally by the long-term survival and expansion of FAH-positive cells of these mice after withdrawal of the rescuing drug NTBC (2-(2-nitro-4-fluoromethylbenzoyl)-1,3-cyclohexanedione). Furthermore, our results demonstrate that both a liver-specific promoter (transthyretin, TTR)-driven FAH transgene and a strong viral promoter (from spleen focus-forming virus, SFFV)-driven FAH transgene rescued the FAH-deficiency phenotypes in the mice derived from the respective gene-corrected iPS cells. In conclusion, our data demonstrate that a lentiviral gene repair strategy does not abrogate the full pluripotent potential of fibroblast-derived iPS cells, and genetic manipulation of iPS cells in combination with tetraploid embryo aggregation provides a practical and rapid approach to evaluate the efficacy of gene correction of human diseases in mouse models.en
dc.language.isoenen
dc.subject.meshAnimalsen
dc.subject.meshCell Survivalen
dc.subject.meshCells, Cultureden
dc.subject.meshChromosomesen
dc.subject.meshCyclohexanonesen
dc.subject.meshDisease Models, Animalen
dc.subject.meshFemaleen
dc.subject.meshFetusen
dc.subject.meshFibroblastsen
dc.subject.meshGene Therapyen
dc.subject.meshGenetic Complementation Testen
dc.subject.meshGenetic Vectorsen
dc.subject.meshHumansen
dc.subject.meshHydrolasesen
dc.subject.meshInduced Pluripotent Stem Cellsen
dc.subject.meshLentivirusen
dc.subject.meshLiveren
dc.subject.meshMiceen
dc.subject.meshMice, Knockouten
dc.subject.meshNitrobenzoatesen
dc.subject.meshPregnancyen
dc.subject.meshPromoter Regions, Geneticen
dc.subject.meshSpleen Focus-Forming Virusesen
dc.subject.meshTetraploidyen
dc.subject.meshTyrosinemiasen
dc.titleGeneration of healthy mice from gene-corrected disease-specific induced pluripotent stem cells.en
dc.typeArticleen
dc.contributor.departmentMax-Planck-Institute for Molecular Biomedicine, Münster, Germany.en
dc.identifier.journalPLoS biologyen

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