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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Autores principales:	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
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	Medicine (General) R5-920 Biochemistry QD415-436
Acceso en línea:	https://doaj.org/article/02dd7fae8580426181008292dcd99f31
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spelling	oai:doaj.org-article:02dd7fae8580426181008292dcd99f312021-11-28T12:06:23ZA familiar study on self-limited childhood epilepsy patients using hIPSC-derived neurons shows a bias towards immaturity at the morphological, electrophysiological and gene expression levels10.1186/s13287-021-02658-21757-6512https://doaj.org/article/02dd7fae8580426181008292dcd99f312021-11-01T00:00:00Zhttps://doi.org/10.1186/s13287-021-02658-2https://doaj.org/toc/1757-6512Abstract 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.Mariana L. CasaliaJuan Cruz CasabonaCorina GarcíaVerónica Cavaliere CandedoHéctor Ramiro QuintáMaría Isabel FaríasJoaquín GonzalezDolores Gonzalez MorónMarta CórdobaDamian ConsalvoGustavo MostoslavskyFrancisco J. UrbanoJuana PasquiniMario Gustavo MurerLorena RelaMarcelo A. KauffmanFernando J. PitossiBMCarticleMedicine (General)R5-920BiochemistryQD415-436ENStem Cell Research & Therapy, Vol 12, Iss 1, Pp 1-16 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine (General) R5-920 Biochemistry QD415-436
spellingShingle	Medicine (General) R5-920 Biochemistry QD415-436 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
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.
format	article
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
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_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_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_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
publisher	BMC
publishDate	2021
url	https://doaj.org/article/02dd7fae8580426181008292dcd99f31
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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

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