Altered cytoskeleton dynamics in patient-derived iPSC-based model of PCDH19 clustering epilepsy
Protocadherin 19 (PCDH19) is an adhesion molecule involved in cell-cell interaction whose mutations cause a drug-resistant form of epilepsy, named PCDH19-Clustering Epilepsy (PCDH19-CE, MIM 300088). The mechanism by which altered PCDH19 function drive pathogenesis is not yet fully understood. Our pr...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2025-01-01
|
| Series: | Frontiers in Cell and Developmental Biology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2024.1518533/full |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1841558703092793344 |
|---|---|
| author | Rossella Borghi Stefania Petrini Valentina Apollonio Marina Trivisano Nicola Specchio Sandra Moreno Enrico Bertini Marco Tartaglia Claudia Compagnucci |
| author_facet | Rossella Borghi Stefania Petrini Valentina Apollonio Marina Trivisano Nicola Specchio Sandra Moreno Enrico Bertini Marco Tartaglia Claudia Compagnucci |
| author_sort | Rossella Borghi |
| collection | DOAJ |
| description | Protocadherin 19 (PCDH19) is an adhesion molecule involved in cell-cell interaction whose mutations cause a drug-resistant form of epilepsy, named PCDH19-Clustering Epilepsy (PCDH19-CE, MIM 300088). The mechanism by which altered PCDH19 function drive pathogenesis is not yet fully understood. Our previous work showed that PCDH19 dysfunction is associated with altered orientation of the mitotic spindle and accelerated neurogenesis, suggesting a contribution of altered cytoskeleton organization in PCDH19-CE pathogenesis in the control of cell division and differentiation. Here, we evaluate the consequences of altered PCDH19 function on microfilaments and microtubules organization, using a disease model obtained from patient-derived induced pluripotent stem cells. We show that iPSC-derived cortical neurons are characterized by altered cytoskeletal dynamics, suggesting that this protocadherin has a role in modulating stability of MFs and MTs. Consistently, the levels of acetylated-tubulin, which is related with stable MTs, are significantly increased in cortical neurons derived from the patient’s iPSCs compared to control cells, supporting the idea that the altered dynamics of the MTs depends on their increased stability. Finally, performing live-imaging experiments using fluorescence recovery after photobleaching and by monitoring GFP-tagged end binding protein 3 (EB3) “comets,” we observe an impairment of the plus-end polymerization speed in PCDH19-mutated cortical neurons, therefore confirming the impaired MT dynamics. In addition to altering the mitotic spindle formation, the present data unveil that PCDH19 dysfunction leads to altered cytoskeletal rearrangement, providing therapeutic targets and pharmacological options to treat this disorder. |
| format | Article |
| id | doaj-art-7a687496018c434ba3af18d10940c042 |
| institution | Kabale University |
| issn | 2296-634X |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj-art-7a687496018c434ba3af18d10940c0422025-01-06T06:59:07ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2025-01-011210.3389/fcell.2024.15185331518533Altered cytoskeleton dynamics in patient-derived iPSC-based model of PCDH19 clustering epilepsyRossella Borghi0Stefania Petrini1Valentina Apollonio2Marina Trivisano3Nicola Specchio4Sandra Moreno5Enrico Bertini6Marco Tartaglia7Claudia Compagnucci8Molecular Genetics and Functional Genomics, Bambino Gesù Children’s Hospital, IRCCS, Rome, ItalyConfocal Microscopy Core Facility, Laboratories, Bambino Gesù, Children’s Research Hospital, IRCCS, Rome, ItalyConfocal Microscopy Core Facility, Laboratories, Bambino Gesù, Children’s Research Hospital, IRCCS, Rome, ItalyNeurology, Epilepsy and Movement Disorders Unit, Bambino Gesù Children’s Hospital, IRCCS, Full Member of European Reference Network EpiCARE, Rome, ItalyNeurology, Epilepsy and Movement Disorders Unit, Bambino Gesù Children’s Hospital, IRCCS, Full Member of European Reference Network EpiCARE, Rome, ItalyDepartment of Science, LIME, University Roma Tre, Rome, ItalyResearch Unit of Neuromuscular and Neurodegenerative Disorders, Bambino Gesù Children’s Hospital, IRCCS, Rome, ItalyMolecular Genetics and Functional Genomics, Bambino Gesù Children’s Hospital, IRCCS, Rome, ItalyMolecular Genetics and Functional Genomics, Bambino Gesù Children’s Hospital, IRCCS, Rome, ItalyProtocadherin 19 (PCDH19) is an adhesion molecule involved in cell-cell interaction whose mutations cause a drug-resistant form of epilepsy, named PCDH19-Clustering Epilepsy (PCDH19-CE, MIM 300088). The mechanism by which altered PCDH19 function drive pathogenesis is not yet fully understood. Our previous work showed that PCDH19 dysfunction is associated with altered orientation of the mitotic spindle and accelerated neurogenesis, suggesting a contribution of altered cytoskeleton organization in PCDH19-CE pathogenesis in the control of cell division and differentiation. Here, we evaluate the consequences of altered PCDH19 function on microfilaments and microtubules organization, using a disease model obtained from patient-derived induced pluripotent stem cells. We show that iPSC-derived cortical neurons are characterized by altered cytoskeletal dynamics, suggesting that this protocadherin has a role in modulating stability of MFs and MTs. Consistently, the levels of acetylated-tubulin, which is related with stable MTs, are significantly increased in cortical neurons derived from the patient’s iPSCs compared to control cells, supporting the idea that the altered dynamics of the MTs depends on their increased stability. Finally, performing live-imaging experiments using fluorescence recovery after photobleaching and by monitoring GFP-tagged end binding protein 3 (EB3) “comets,” we observe an impairment of the plus-end polymerization speed in PCDH19-mutated cortical neurons, therefore confirming the impaired MT dynamics. In addition to altering the mitotic spindle formation, the present data unveil that PCDH19 dysfunction leads to altered cytoskeletal rearrangement, providing therapeutic targets and pharmacological options to treat this disorder.https://www.frontiersin.org/articles/10.3389/fcell.2024.1518533/fullPCDH19epilepsyIPSC-derived neuronsmicrofilamentsmicrotubulescytoskeletal dynamics |
| spellingShingle | Rossella Borghi Stefania Petrini Valentina Apollonio Marina Trivisano Nicola Specchio Sandra Moreno Enrico Bertini Marco Tartaglia Claudia Compagnucci Altered cytoskeleton dynamics in patient-derived iPSC-based model of PCDH19 clustering epilepsy Frontiers in Cell and Developmental Biology PCDH19 epilepsy IPSC-derived neurons microfilaments microtubules cytoskeletal dynamics |
| title | Altered cytoskeleton dynamics in patient-derived iPSC-based model of PCDH19 clustering epilepsy |
| title_full | Altered cytoskeleton dynamics in patient-derived iPSC-based model of PCDH19 clustering epilepsy |
| title_fullStr | Altered cytoskeleton dynamics in patient-derived iPSC-based model of PCDH19 clustering epilepsy |
| title_full_unstemmed | Altered cytoskeleton dynamics in patient-derived iPSC-based model of PCDH19 clustering epilepsy |
| title_short | Altered cytoskeleton dynamics in patient-derived iPSC-based model of PCDH19 clustering epilepsy |
| title_sort | altered cytoskeleton dynamics in patient derived ipsc based model of pcdh19 clustering epilepsy |
| topic | PCDH19 epilepsy IPSC-derived neurons microfilaments microtubules cytoskeletal dynamics |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2024.1518533/full |
| work_keys_str_mv | AT rossellaborghi alteredcytoskeletondynamicsinpatientderivedipscbasedmodelofpcdh19clusteringepilepsy AT stefaniapetrini alteredcytoskeletondynamicsinpatientderivedipscbasedmodelofpcdh19clusteringepilepsy AT valentinaapollonio alteredcytoskeletondynamicsinpatientderivedipscbasedmodelofpcdh19clusteringepilepsy AT marinatrivisano alteredcytoskeletondynamicsinpatientderivedipscbasedmodelofpcdh19clusteringepilepsy AT nicolaspecchio alteredcytoskeletondynamicsinpatientderivedipscbasedmodelofpcdh19clusteringepilepsy AT sandramoreno alteredcytoskeletondynamicsinpatientderivedipscbasedmodelofpcdh19clusteringepilepsy AT enricobertini alteredcytoskeletondynamicsinpatientderivedipscbasedmodelofpcdh19clusteringepilepsy AT marcotartaglia alteredcytoskeletondynamicsinpatientderivedipscbasedmodelofpcdh19clusteringepilepsy AT claudiacompagnucci alteredcytoskeletondynamicsinpatientderivedipscbasedmodelofpcdh19clusteringepilepsy |