The Generation of Human iPSC Lines from Three Individuals with Dravet Syndrome and Characterization of Neural Differentiation Markers in iPSC-Derived Ventral Forebrain Organoid Model
Dravet syndrome (DRVT) is a rare form of neurodevelopmental disorder with a high risk of sudden unexpected death in epilepsy (SUDEP), caused mainly (>80% cases) by mutations in the <i>SCN1A</i> gene, coding the Nav1.1 protein (alfa-subunit of voltage-sensitive sodium channel). Mutatio...
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MDPI AG
2023-01-01
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| Online Access: | https://www.mdpi.com/2073-4409/12/2/339 |
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| author | Valery Zayat Zuzanna Kuczynska Michal Liput Erkan Metin Sylwia Rzonca-Niewczas Marta Smyk Tomasz Mazurczak Alicja Goszczanska-Ciuchta Pawel Leszczynski Dorota Hoffman-Zacharska Leonora Buzanska |
| author_facet | Valery Zayat Zuzanna Kuczynska Michal Liput Erkan Metin Sylwia Rzonca-Niewczas Marta Smyk Tomasz Mazurczak Alicja Goszczanska-Ciuchta Pawel Leszczynski Dorota Hoffman-Zacharska Leonora Buzanska |
| author_sort | Valery Zayat |
| collection | DOAJ |
| description | Dravet syndrome (DRVT) is a rare form of neurodevelopmental disorder with a high risk of sudden unexpected death in epilepsy (SUDEP), caused mainly (>80% cases) by mutations in the <i>SCN1A</i> gene, coding the Nav1.1 protein (alfa-subunit of voltage-sensitive sodium channel). Mutations in <i>SCN1A</i> are linked to heterogenous epileptic phenotypes of various types, severity, and patient prognosis. Here we generated iPSC lines from fibroblasts obtained from three individuals affected with DRVT carrying distinct mutations in the <i>SCN1A</i> gene (nonsense mutation p.Ser1516*, missense mutation p.Arg1596His, and splicing mutation c.2589+2dupT). The iPSC lines, generated with the non-integrative approach, retained the distinct <i>SCN1A</i> gene mutation of the donor fibroblasts and were characterized by confirming the expression of the pluripotency markers, the three-germ layer differentiation potential, the absence of exogenous vector expression, and a normal karyotype. The generated iPSC lines were used to establish ventral forebrain organoids, the most affected type of neurons in the pathology of DRVT. The DRVT organoid model will provide an additional resource for deciphering the pathology behind Nav1.1 haploinsufficiency and drug screening to remediate the functional deficits associated with the disease. |
| first_indexed | 2024-03-09T13:12:09Z |
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| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-09T13:12:09Z |
| publishDate | 2023-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-fda844ebb4364affb5ff16e68a9a32312023-11-30T21:41:06ZengMDPI AGCells2073-44092023-01-0112233910.3390/cells12020339The Generation of Human iPSC Lines from Three Individuals with Dravet Syndrome and Characterization of Neural Differentiation Markers in iPSC-Derived Ventral Forebrain Organoid ModelValery Zayat0Zuzanna Kuczynska1Michal Liput2Erkan Metin3Sylwia Rzonca-Niewczas4Marta Smyk5Tomasz Mazurczak6Alicja Goszczanska-Ciuchta7Pawel Leszczynski8Dorota Hoffman-Zacharska9Leonora Buzanska10Department of Stem Cell Bioengineering, Mossakowski Medical Research Institute, Polish Academy of Sciences, 02-106 Warsaw, PolandDepartment of Stem Cell Bioengineering, Mossakowski Medical Research Institute, Polish Academy of Sciences, 02-106 Warsaw, PolandDepartment of Stem Cell Bioengineering, Mossakowski Medical Research Institute, Polish Academy of Sciences, 02-106 Warsaw, PolandDepartment of Stem Cell Bioengineering, Mossakowski Medical Research Institute, Polish Academy of Sciences, 02-106 Warsaw, PolandMedical Genetics Department, Institute of Mother and Child, 01-211 Warsaw, PolandMedical Genetics Department, Institute of Mother and Child, 01-211 Warsaw, PolandMedical Genetics Department, Institute of Mother and Child, 01-211 Warsaw, PolandMedical Genetics Department, Institute of Mother and Child, 01-211 Warsaw, PolandDepartment of Stem Cell Bioengineering, Mossakowski Medical Research Institute, Polish Academy of Sciences, 02-106 Warsaw, PolandMedical Genetics Department, Institute of Mother and Child, 01-211 Warsaw, PolandDepartment of Stem Cell Bioengineering, Mossakowski Medical Research Institute, Polish Academy of Sciences, 02-106 Warsaw, PolandDravet syndrome (DRVT) is a rare form of neurodevelopmental disorder with a high risk of sudden unexpected death in epilepsy (SUDEP), caused mainly (>80% cases) by mutations in the <i>SCN1A</i> gene, coding the Nav1.1 protein (alfa-subunit of voltage-sensitive sodium channel). Mutations in <i>SCN1A</i> are linked to heterogenous epileptic phenotypes of various types, severity, and patient prognosis. Here we generated iPSC lines from fibroblasts obtained from three individuals affected with DRVT carrying distinct mutations in the <i>SCN1A</i> gene (nonsense mutation p.Ser1516*, missense mutation p.Arg1596His, and splicing mutation c.2589+2dupT). The iPSC lines, generated with the non-integrative approach, retained the distinct <i>SCN1A</i> gene mutation of the donor fibroblasts and were characterized by confirming the expression of the pluripotency markers, the three-germ layer differentiation potential, the absence of exogenous vector expression, and a normal karyotype. The generated iPSC lines were used to establish ventral forebrain organoids, the most affected type of neurons in the pathology of DRVT. The DRVT organoid model will provide an additional resource for deciphering the pathology behind Nav1.1 haploinsufficiency and drug screening to remediate the functional deficits associated with the disease.https://www.mdpi.com/2073-4409/12/2/339<i>SCN1A</i>-related disordersDravet syndromePanayiotopoulos syndromeNav1.1 haploinsufficiencystem cell reprogrammingorganoids |
| spellingShingle | Valery Zayat Zuzanna Kuczynska Michal Liput Erkan Metin Sylwia Rzonca-Niewczas Marta Smyk Tomasz Mazurczak Alicja Goszczanska-Ciuchta Pawel Leszczynski Dorota Hoffman-Zacharska Leonora Buzanska The Generation of Human iPSC Lines from Three Individuals with Dravet Syndrome and Characterization of Neural Differentiation Markers in iPSC-Derived Ventral Forebrain Organoid Model Cells <i>SCN1A</i>-related disorders Dravet syndrome Panayiotopoulos syndrome Nav1.1 haploinsufficiency stem cell reprogramming organoids |
| title | The Generation of Human iPSC Lines from Three Individuals with Dravet Syndrome and Characterization of Neural Differentiation Markers in iPSC-Derived Ventral Forebrain Organoid Model |
| title_full | The Generation of Human iPSC Lines from Three Individuals with Dravet Syndrome and Characterization of Neural Differentiation Markers in iPSC-Derived Ventral Forebrain Organoid Model |
| title_fullStr | The Generation of Human iPSC Lines from Three Individuals with Dravet Syndrome and Characterization of Neural Differentiation Markers in iPSC-Derived Ventral Forebrain Organoid Model |
| title_full_unstemmed | The Generation of Human iPSC Lines from Three Individuals with Dravet Syndrome and Characterization of Neural Differentiation Markers in iPSC-Derived Ventral Forebrain Organoid Model |
| title_short | The Generation of Human iPSC Lines from Three Individuals with Dravet Syndrome and Characterization of Neural Differentiation Markers in iPSC-Derived Ventral Forebrain Organoid Model |
| title_sort | generation of human ipsc lines from three individuals with dravet syndrome and characterization of neural differentiation markers in ipsc derived ventral forebrain organoid model |
| topic | <i>SCN1A</i>-related disorders Dravet syndrome Panayiotopoulos syndrome Nav1.1 haploinsufficiency stem cell reprogramming organoids |
| url | https://www.mdpi.com/2073-4409/12/2/339 |
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