A humanized yeast model for studying TRAPP complex mutations; proof-of-concept using variants from an individual with a TRAPPC1-associated neurodevelopmental syndrome

Variants in membrane trafficking proteins are known to cause rare disorders with severe symptoms. The highly conserved transport protein particle (TRAPP) complexes are key membrane trafficking regulators that are also involved in autophagy. Pathogenic genetic variants in specific TRAPP subunits are...

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Autores: Zykaj, Erta, Abboud, Chelsea, Asadi, Paria, Warsame, Simane, Almousa, Hashem, Milev, Miroslav P., Greco, Brittany M., López Sánchez, Marcos, 1986-, Bratkovic, Drago, Kachroo, Aashiq H., Pérez Jurado, Luis Alberto, Sacher, Michael
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2024
País:España
Institución:Universitat Pompeu Fabra
Repositorio:Repositorio Digital de la UPF
OAI Identifier:oai:repositori.upf.edu:10230/68491
Acceso en línea:http://hdl.handle.net/10230/68491
http://dx.doi.org/10.3390/cells13171457
Access Level:acceso abierto
Palabra clave:Golgi
TRAPP
TRAPPC1
Autophagy
Humanization
Mutation
Yeast
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spelling A humanized yeast model for studying TRAPP complex mutations; proof-of-concept using variants from an individual with a TRAPPC1-associated neurodevelopmental syndromeZykaj, ErtaAbboud, ChelseaAsadi, PariaWarsame, SimaneAlmousa, HashemMilev, Miroslav P.Greco, Brittany M.López Sánchez, Marcos, 1986-Bratkovic, DragoKachroo, Aashiq H.Pérez Jurado, Luis AlbertoSacher, MichaelGolgiTRAPPTRAPPC1AutophagyHumanizationMutationYeastVariants in membrane trafficking proteins are known to cause rare disorders with severe symptoms. The highly conserved transport protein particle (TRAPP) complexes are key membrane trafficking regulators that are also involved in autophagy. Pathogenic genetic variants in specific TRAPP subunits are linked to neurological disorders, muscular dystrophies, and skeletal dysplasias. Characterizing these variants and their phenotypes is important for understanding the general and specialized roles of TRAPP subunits as well as for patient diagnosis. Patient-derived cells are not always available, which poses a limitation for the study of these diseases. Therefore, other systems, like the yeast Saccharomyces cerevisiae, can be used to dissect the mechanisms at the intracellular level underlying these disorders. The development of CRISPR/Cas9 technology in yeast has enabled a scar-less editing method that creates an efficient humanized yeast model. In this study, core yeast subunits were humanized by replacing them with their human orthologs, and TRAPPC1, TRAPPC2, TRAPPC2L, TRAPPC6A, and TRAPPC6B were found to successfully replace their yeast counterparts. This system was used for studying the first reported individual with an autosomal recessive disorder caused by biallelic TRAPPC1 variants, a girl with a severe neurodevelopmental disorder and myopathy. We show that the maternal variant (TRAPPC1 p.(Val121Alafs*3)) is non-functional while the paternal variant (TRAPPC1 p.(His22_Lys24del)) is conditional-lethal and affects secretion and non-selective autophagy in yeast. This parallels defects seen in fibroblasts derived from this individual which also showed membrane trafficking defects and altered Golgi morphology, all of which were rescued in the human system by wild-type TRAPPC1. This study suggests that humanized yeast can be an efficient means to study TRAPP subunit variants in the absence of human cells and can assign significance to variants of unknown significance (VUS). This study lays the foundation for characterizing further TRAPP variants through this system, rapidly contributing to disease diagnosis.MDPI202420242024info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/pdfhttp://hdl.handle.net/10230/68491http://dx.doi.org/10.3390/cells13171457reponame:Repositorio Digital de la UPFinstname:Universitat Pompeu FabraInglésCells. 2024 Aug 30;13(17):1457© 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).http://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:repositori.upf.edu:10230/684912026-06-12T07:21:37Z
dc.title.none.fl_str_mv A humanized yeast model for studying TRAPP complex mutations; proof-of-concept using variants from an individual with a TRAPPC1-associated neurodevelopmental syndrome
title A humanized yeast model for studying TRAPP complex mutations; proof-of-concept using variants from an individual with a TRAPPC1-associated neurodevelopmental syndrome
spellingShingle A humanized yeast model for studying TRAPP complex mutations; proof-of-concept using variants from an individual with a TRAPPC1-associated neurodevelopmental syndrome
Zykaj, Erta
Golgi
TRAPP
TRAPPC1
Autophagy
Humanization
Mutation
Yeast
title_short A humanized yeast model for studying TRAPP complex mutations; proof-of-concept using variants from an individual with a TRAPPC1-associated neurodevelopmental syndrome
title_full A humanized yeast model for studying TRAPP complex mutations; proof-of-concept using variants from an individual with a TRAPPC1-associated neurodevelopmental syndrome
title_fullStr A humanized yeast model for studying TRAPP complex mutations; proof-of-concept using variants from an individual with a TRAPPC1-associated neurodevelopmental syndrome
title_full_unstemmed A humanized yeast model for studying TRAPP complex mutations; proof-of-concept using variants from an individual with a TRAPPC1-associated neurodevelopmental syndrome
title_sort A humanized yeast model for studying TRAPP complex mutations; proof-of-concept using variants from an individual with a TRAPPC1-associated neurodevelopmental syndrome
dc.creator.none.fl_str_mv Zykaj, Erta
Abboud, Chelsea
Asadi, Paria
Warsame, Simane
Almousa, Hashem
Milev, Miroslav P.
Greco, Brittany M.
López Sánchez, Marcos, 1986-
Bratkovic, Drago
Kachroo, Aashiq H.
Pérez Jurado, Luis Alberto
Sacher, Michael
author Zykaj, Erta
author_facet Zykaj, Erta
Abboud, Chelsea
Asadi, Paria
Warsame, Simane
Almousa, Hashem
Milev, Miroslav P.
Greco, Brittany M.
López Sánchez, Marcos, 1986-
Bratkovic, Drago
Kachroo, Aashiq H.
Pérez Jurado, Luis Alberto
Sacher, Michael
author_role author
author2 Abboud, Chelsea
Asadi, Paria
Warsame, Simane
Almousa, Hashem
Milev, Miroslav P.
Greco, Brittany M.
López Sánchez, Marcos, 1986-
Bratkovic, Drago
Kachroo, Aashiq H.
Pérez Jurado, Luis Alberto
Sacher, Michael
author2_role author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Golgi
TRAPP
TRAPPC1
Autophagy
Humanization
Mutation
Yeast
topic Golgi
TRAPP
TRAPPC1
Autophagy
Humanization
Mutation
Yeast
description Variants in membrane trafficking proteins are known to cause rare disorders with severe symptoms. The highly conserved transport protein particle (TRAPP) complexes are key membrane trafficking regulators that are also involved in autophagy. Pathogenic genetic variants in specific TRAPP subunits are linked to neurological disorders, muscular dystrophies, and skeletal dysplasias. Characterizing these variants and their phenotypes is important for understanding the general and specialized roles of TRAPP subunits as well as for patient diagnosis. Patient-derived cells are not always available, which poses a limitation for the study of these diseases. Therefore, other systems, like the yeast Saccharomyces cerevisiae, can be used to dissect the mechanisms at the intracellular level underlying these disorders. The development of CRISPR/Cas9 technology in yeast has enabled a scar-less editing method that creates an efficient humanized yeast model. In this study, core yeast subunits were humanized by replacing them with their human orthologs, and TRAPPC1, TRAPPC2, TRAPPC2L, TRAPPC6A, and TRAPPC6B were found to successfully replace their yeast counterparts. This system was used for studying the first reported individual with an autosomal recessive disorder caused by biallelic TRAPPC1 variants, a girl with a severe neurodevelopmental disorder and myopathy. We show that the maternal variant (TRAPPC1 p.(Val121Alafs*3)) is non-functional while the paternal variant (TRAPPC1 p.(His22_Lys24del)) is conditional-lethal and affects secretion and non-selective autophagy in yeast. This parallels defects seen in fibroblasts derived from this individual which also showed membrane trafficking defects and altered Golgi morphology, all of which were rescued in the human system by wild-type TRAPPC1. This study suggests that humanized yeast can be an efficient means to study TRAPP subunit variants in the absence of human cells and can assign significance to variants of unknown significance (VUS). This study lays the foundation for characterizing further TRAPP variants through this system, rapidly contributing to disease diagnosis.
publishDate 2024
dc.date.none.fl_str_mv 2024
2024
2024
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10230/68491
http://dx.doi.org/10.3390/cells13171457
url http://hdl.handle.net/10230/68491
http://dx.doi.org/10.3390/cells13171457
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Cells. 2024 Aug 30;13(17):1457
dc.rights.none.fl_str_mv http://creativecommons.org/licenses/by/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv reponame:Repositorio Digital de la UPF
instname:Universitat Pompeu Fabra
instname_str Universitat Pompeu Fabra
reponame_str Repositorio Digital de la UPF
collection Repositorio Digital de la UPF
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,812429

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