A familiar study on self-limited childhood epilepsy patients using hIPSC-derived neurons shows a bias towards immaturity at the morphological, electrophysiological and gene expression levels
Abstract Background Self-limited Childhood Epilepsies are the most prevalent epileptic syndrome in children. Its pathogenesis is unknown. In this disease, symptoms resolve spontaneously in approximately 50% of patients when maturity is reached, prompting to a maturation problem. The purpose of this...
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| Format: | Article |
| Language: | English |
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BMC
2021-11-01
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| Series: | Stem Cell Research & Therapy |
| Online Access: | https://doi.org/10.1186/s13287-021-02658-2 |
| _version_ | 1830287586957983744 |
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| author | Mariana L. Casalia Juan Cruz Casabona Corina García Verónica Cavaliere Candedo Héctor Ramiro Quintá María Isabel Farías Joaquín Gonzalez Dolores Gonzalez Morón Marta Córdoba Damian Consalvo Gustavo Mostoslavsky Francisco J. Urbano Juana Pasquini Mario Gustavo Murer Lorena Rela Marcelo A. Kauffman Fernando J. Pitossi |
| author_facet | Mariana L. Casalia Juan Cruz Casabona Corina García Verónica Cavaliere Candedo Héctor Ramiro Quintá María Isabel Farías Joaquín Gonzalez Dolores Gonzalez Morón Marta Córdoba Damian Consalvo Gustavo Mostoslavsky Francisco J. Urbano Juana Pasquini Mario Gustavo Murer Lorena Rela Marcelo A. Kauffman Fernando J. Pitossi |
| author_sort | Mariana L. Casalia |
| collection | DOAJ |
| description | Abstract Background Self-limited Childhood Epilepsies are the most prevalent epileptic syndrome in children. Its pathogenesis is unknown. In this disease, symptoms resolve spontaneously in approximately 50% of patients when maturity is reached, prompting to a maturation problem. The purpose of this study was to understand the molecular bases of this disease by generating and analyzing induced pluripotent stem cell-derived neurons from a family with 7 siblings, among whom 4 suffer from this disease. Methods Two affected siblings and, as controls, a healthy sister and the unaffected mother of the family were studied. Using exome sequencing, a homozygous variant in the FYVE, RhoGEF and PH Domain Containing 6 gene was identified in the patients as a putative genetic factor that could contribute to the development of this familial disorder. After informed consent was signed, skin biopsies from the 4 individuals were collected, fibroblasts were derived and reprogrammed and neurons were generated and characterized by markers and electrophysiology. Morphological, electrophysiological and gene expression analyses were performed on these neurons. Results Bona fide induced pluripotent stem cells and derived neurons could be generated in all cases. Overall, there were no major shifts in neuronal marker expression among patient and control-derived neurons. Compared to two familial controls, neurons from patients showed shorter axonal length, a dramatic reduction in synapsin-1 levels and cytoskeleton disorganization. In addition, neurons from patients developed a lower action potential threshold with time of in vitro differentiation and the amount of current needed to elicit an action potential (rheobase) was smaller in cells recorded from NE derived from patients at 12 weeks of differentiation when compared with shorter times in culture. These results indicate an increased excitability in patient cells that emerges with the time in culture. Finally, functional genomic analysis showed a biased towards immaturity in patient-derived neurons. Conclusions We are reporting the first in vitro model of self-limited childhood epilepsy, providing the cellular bases for future in-depth studies to understand its pathogenesis. Our results show patient-specific neuronal features reflecting immaturity, in resonance with the course of the disease and previous imaging studies. |
| first_indexed | 2024-12-19T04:20:31Z |
| format | Article |
| id | doaj.art-02dd7fae8580426181008292dcd99f31 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-19T04:20:31Z |
| publishDate | 2021-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-02dd7fae8580426181008292dcd99f312022-12-21T20:36:10ZengBMCStem Cell Research & Therapy1757-65122021-11-0112111610.1186/s13287-021-02658-2A familiar study on self-limited childhood epilepsy patients using hIPSC-derived neurons shows a bias towards immaturity at the morphological, electrophysiological and gene expression levelsMariana L. Casalia0Juan Cruz Casabona1Corina García2Verónica Cavaliere Candedo3Héctor Ramiro Quintá4María Isabel Farías5Joaquín Gonzalez6Dolores Gonzalez Morón7Marta Córdoba8Damian Consalvo9Gustavo Mostoslavsky10Francisco J. Urbano11Juana Pasquini12Mario Gustavo Murer13Lorena Rela14Marcelo A. Kauffman15Fernando J. Pitossi16Institute Leloir Foundation- IIBBA-CONICETInstitute Leloir Foundation- IIBBA-CONICETInstitute Leloir Foundation- IIBBA-CONICETInstitute Leloir Foundation- IIBBA-CONICETCONICET and Laboratorio de Medicina Experimental “Dr. J Toblli”, Hospital AlemánInstitute Leloir Foundation- IIBBA-CONICETInstitute Leloir Foundation- IIBBA-CONICETConsultorio y Laboratorio de Neurogenética, Centro Universitario de Neurología “José María Ramos Mejía” Facultad de Medicina, UBA & Instituto de Investigaciones en Medicina Traslacional, Facultad de Ciencias Biomédicas, Universidad Austral-CONICETConsultorio y Laboratorio de Neurogenética, Centro Universitario de Neurología “José María Ramos Mejía” Facultad de Medicina, UBA & Instituto de Investigaciones en Medicina Traslacional, Facultad de Ciencias Biomédicas, Universidad Austral-CONICETConsultorio y Laboratorio de Neurogenética, Centro Universitario de Neurología “José María Ramos Mejía” Facultad de Medicina, UBA & Instituto de Investigaciones en Medicina Traslacional, Facultad de Ciencias Biomédicas, Universidad Austral-CONICETCenter For Regenerative Medicine (CReM) of Boston University and Boston Medical CenterDepartment of Physiology, Molecular and Cellular Biology “Dr. Héctor Maldonado”, Faculty of Exact and Natural Sciences, University of Buenos Aires, IFIBYNE-CONICETFaculty of Pharmacy and Biochemistry, University of Buenos AiresUniversidad de Buenos Aires, Facultad de Medicina, Departamento de Ciencias Fisiológicas, Grupo de Neurociencia de SistemasUniversidad de Buenos Aires, Facultad de Medicina, Departamento de Ciencias Fisiológicas, Grupo de Neurociencia de SistemasConsultorio y Laboratorio de Neurogenética, Centro Universitario de Neurología “José María Ramos Mejía” Facultad de Medicina, UBA & Instituto de Investigaciones en Medicina Traslacional, Facultad de Ciencias Biomédicas, Universidad Austral-CONICETInstitute Leloir Foundation- IIBBA-CONICETAbstract Background Self-limited Childhood Epilepsies are the most prevalent epileptic syndrome in children. Its pathogenesis is unknown. In this disease, symptoms resolve spontaneously in approximately 50% of patients when maturity is reached, prompting to a maturation problem. The purpose of this study was to understand the molecular bases of this disease by generating and analyzing induced pluripotent stem cell-derived neurons from a family with 7 siblings, among whom 4 suffer from this disease. Methods Two affected siblings and, as controls, a healthy sister and the unaffected mother of the family were studied. Using exome sequencing, a homozygous variant in the FYVE, RhoGEF and PH Domain Containing 6 gene was identified in the patients as a putative genetic factor that could contribute to the development of this familial disorder. After informed consent was signed, skin biopsies from the 4 individuals were collected, fibroblasts were derived and reprogrammed and neurons were generated and characterized by markers and electrophysiology. Morphological, electrophysiological and gene expression analyses were performed on these neurons. Results Bona fide induced pluripotent stem cells and derived neurons could be generated in all cases. Overall, there were no major shifts in neuronal marker expression among patient and control-derived neurons. Compared to two familial controls, neurons from patients showed shorter axonal length, a dramatic reduction in synapsin-1 levels and cytoskeleton disorganization. In addition, neurons from patients developed a lower action potential threshold with time of in vitro differentiation and the amount of current needed to elicit an action potential (rheobase) was smaller in cells recorded from NE derived from patients at 12 weeks of differentiation when compared with shorter times in culture. These results indicate an increased excitability in patient cells that emerges with the time in culture. Finally, functional genomic analysis showed a biased towards immaturity in patient-derived neurons. Conclusions We are reporting the first in vitro model of self-limited childhood epilepsy, providing the cellular bases for future in-depth studies to understand its pathogenesis. Our results show patient-specific neuronal features reflecting immaturity, in resonance with the course of the disease and previous imaging studies.https://doi.org/10.1186/s13287-021-02658-2 |
| spellingShingle | Mariana L. Casalia Juan Cruz Casabona Corina García Verónica Cavaliere Candedo Héctor Ramiro Quintá María Isabel Farías Joaquín Gonzalez Dolores Gonzalez Morón Marta Córdoba Damian Consalvo Gustavo Mostoslavsky Francisco J. Urbano Juana Pasquini Mario Gustavo Murer Lorena Rela Marcelo A. Kauffman Fernando J. Pitossi A familiar study on self-limited childhood epilepsy patients using hIPSC-derived neurons shows a bias towards immaturity at the morphological, electrophysiological and gene expression levels Stem Cell Research & Therapy |
| title | A familiar study on self-limited childhood epilepsy patients using hIPSC-derived neurons shows a bias towards immaturity at the morphological, electrophysiological and gene expression levels |
| title_full | A familiar study on self-limited childhood epilepsy patients using hIPSC-derived neurons shows a bias towards immaturity at the morphological, electrophysiological and gene expression levels |
| title_fullStr | A familiar study on self-limited childhood epilepsy patients using hIPSC-derived neurons shows a bias towards immaturity at the morphological, electrophysiological and gene expression levels |
| title_full_unstemmed | A familiar study on self-limited childhood epilepsy patients using hIPSC-derived neurons shows a bias towards immaturity at the morphological, electrophysiological and gene expression levels |
| title_short | A familiar study on self-limited childhood epilepsy patients using hIPSC-derived neurons shows a bias towards immaturity at the morphological, electrophysiological and gene expression levels |
| title_sort | familiar study on self limited childhood epilepsy patients using hipsc derived neurons shows a bias towards immaturity at the morphological electrophysiological and gene expression levels |
| url | https://doi.org/10.1186/s13287-021-02658-2 |
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