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

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• dc.contributor.author Zykaj, Erta
• dc.contributor.author Abboud, Chelsea
• dc.contributor.author Asadi, Paria
• dc.contributor.author Warsame, Simane
• dc.contributor.author Almousa, Hashem
• dc.contributor.author Milev, Miroslav P.
• dc.contributor.author Greco, Brittany M.
• dc.contributor.author López Sánchez, Marcos, 1986-
• dc.contributor.author Bratkovic, Drago
• dc.contributor.author Kachroo, Aashiq H.
• dc.contributor.author Pérez Jurado, Luis Alberto
• dc.contributor.author Sacher, Michael
• dc.date.accessioned 2024-11-12T06:36:35Z
• dc.date.available 2024-11-12T06:36:35Z
• dc.date.issued 2024
• dc.description.abstract 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.
• dc.format.mimetype application/pdf
• dc.identifier.citation Zykaj E, Abboud C, Asadi P, Warsame S, Almousa H, Milev MP, et al. A humanized yeast model for studying TRAPP complex mutations; proof-of-concept using variants from an individual with a TRAPPC1-associated neurodevelopmental syndrome. Cells. 2024 Aug 30;13(17):1457. DOI: 10.3390/cells13171457
• dc.identifier.doi http://dx.doi.org/10.3390/cells13171457
• dc.identifier.issn 2073-4409
• dc.identifier.uri http://hdl.handle.net/10230/68491
• dc.language.iso eng
• dc.publisher MDPI
• dc.relation.ispartof Cells. 2024 Aug 30;13(17):1457
• dc.rights © 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/).
• dc.rights.accessRights info:eu-repo/semantics/openAccess
• dc.rights.uri http://creativecommons.org/licenses/by/4.0/
• dc.subject.keyword Golgi
• dc.subject.keyword TRAPP
• dc.subject.keyword TRAPPC1
• dc.subject.keyword Autophagy
• dc.subject.keyword Humanization
• dc.subject.keyword Mutation
• dc.subject.keyword Yeast
• dc.title A humanized yeast model for studying TRAPP complex mutations; proof-of-concept using variants from an individual with a TRAPPC1-associated neurodevelopmental syndrome
• dc.type info:eu-repo/semantics/article
• dc.type.version info:eu-repo/semantics/publishedVersion