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Pathogenic TRIO variants associated with neurodevelopmental disorders perturb the molecular regulation of TRIO and axon pathfinding in vivo

Pathogenic TRIO variants associated with neurodevelopmental disorders perturb the molecular regulation of TRIO and axon pathfinding in vivo
Pathogenic TRIO variants associated with neurodevelopmental disorders perturb the molecular regulation of TRIO and axon pathfinding in vivo
The RhoGEF TRIO is known to play a major role in neuronal development by controlling actin cytoskeleton remodeling, primarily through the activation of the RAC1 GTPase. Numerous de novo mutations in the TRIO gene have been identified in individuals with neurodevelopmental disorders (NDDs). We have previously established the first phenotype/genotype correlation in TRIO-associated diseases, with striking correlation between the clinical features of the individuals and the opposite modulation of RAC1 activity by TRIO variants targeting different domains. The mutations hyperactivating RAC1 are of particular interest, as they are recurrently found in patients and are associated with a severe form of NDD and macrocephaly, indicating their importance in the etiology of the disease. Yet, it remains unknown how these pathogenic TRIO variants disrupt TRIO activity at a molecular level and how they affect neurodevelopmental processes such as axon outgrowth or guidance. Here we report an additional cohort of individuals carrying a pathogenic TRIO variant that reinforces our initial phenotype/genotype correlation. More importantly, by performing conformation predictions coupled to biochemical validation, we propose a model whereby TRIO is inhibited by an intramolecular fold and NDD-associated variants relieve this inhibition, leading to RAC1 hyperactivation. Moreover, we show that in cultured primary neurons and in the zebrafish developmental model, these gain-of-function variants differentially affect axon outgrowth and branching in vitro and in vivo, as compared to loss-of-function TRIO variants. In summary, by combining clinical, molecular, cellular and in vivo data, we provide compelling new evidence for the pathogenicity of novel genetic variants targeting the TRIO gene in NDDs. We report a novel mechanism whereby the fine-tuned regulation of TRIO activity is critical for proper neuronal development and is disrupted by pathogenic mutations.
1359-4184
1527-1544
Bonnet, Maxime
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Roche, Fiona
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Fagotto-Kaufmann, Christine
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Gazdagh, Gabriella
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Truong, Iona
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Comunale, Franck
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Barbosa, Sonia
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Bonhomme, Marion
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Nafati, Nicholas
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Hunt, David
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Rodriguez, Monserrat Pons
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Chaudhry, Ayeshah
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Shears, Deborah
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Madruga, Marcos
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Vansenne, Fleur
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Curie, Aurore
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Kajava, Andry V
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Baralle, Diana
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Fassier, Coralie
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Debant, Anne
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Schmidt, Susanne
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Bonnet, Maxime
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Roche, Fiona
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Fagotto-Kaufmann, Christine
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Gazdagh, Gabriella
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Truong, Iona
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Comunale, Franck
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Barbosa, Sonia
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Bonhomme, Marion
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Nafati, Nicholas
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Hunt, David
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Rodriguez, Monserrat Pons
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Chaudhry, Ayeshah
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Shears, Deborah
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Madruga, Marcos
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Vansenne, Fleur
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Curie, Aurore
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Kajava, Andry V
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Baralle, Diana
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Fassier, Coralie
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Debant, Anne
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Schmidt, Susanne
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Bonnet, Maxime, Roche, Fiona, Fagotto-Kaufmann, Christine, Gazdagh, Gabriella, Truong, Iona, Comunale, Franck, Barbosa, Sonia, Bonhomme, Marion, Nafati, Nicholas, Hunt, David, Rodriguez, Monserrat Pons, Chaudhry, Ayeshah, Shears, Deborah, Madruga, Marcos, Vansenne, Fleur, Curie, Aurore, Kajava, Andry V, Baralle, Diana, Fassier, Coralie, Debant, Anne and Schmidt, Susanne (2023) Pathogenic TRIO variants associated with neurodevelopmental disorders perturb the molecular regulation of TRIO and axon pathfinding in vivo. Molecular Psychiatry, 28 (4), 1527-1544. (doi:10.1038/s41380-023-01963-x).

Record type: Article

Abstract

The RhoGEF TRIO is known to play a major role in neuronal development by controlling actin cytoskeleton remodeling, primarily through the activation of the RAC1 GTPase. Numerous de novo mutations in the TRIO gene have been identified in individuals with neurodevelopmental disorders (NDDs). We have previously established the first phenotype/genotype correlation in TRIO-associated diseases, with striking correlation between the clinical features of the individuals and the opposite modulation of RAC1 activity by TRIO variants targeting different domains. The mutations hyperactivating RAC1 are of particular interest, as they are recurrently found in patients and are associated with a severe form of NDD and macrocephaly, indicating their importance in the etiology of the disease. Yet, it remains unknown how these pathogenic TRIO variants disrupt TRIO activity at a molecular level and how they affect neurodevelopmental processes such as axon outgrowth or guidance. Here we report an additional cohort of individuals carrying a pathogenic TRIO variant that reinforces our initial phenotype/genotype correlation. More importantly, by performing conformation predictions coupled to biochemical validation, we propose a model whereby TRIO is inhibited by an intramolecular fold and NDD-associated variants relieve this inhibition, leading to RAC1 hyperactivation. Moreover, we show that in cultured primary neurons and in the zebrafish developmental model, these gain-of-function variants differentially affect axon outgrowth and branching in vitro and in vivo, as compared to loss-of-function TRIO variants. In summary, by combining clinical, molecular, cellular and in vivo data, we provide compelling new evidence for the pathogenicity of novel genetic variants targeting the TRIO gene in NDDs. We report a novel mechanism whereby the fine-tuned regulation of TRIO activity is critical for proper neuronal development and is disrupted by pathogenic mutations.

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Accepted/In Press date: 13 January 2023
e-pub ahead of print date: 30 January 2023
Published date: 1 April 2023
Additional Information: Funding Information: We thank all members of the Debant team as well as Xavier Nicol (Institut de la Vision, Paris) for helpful discussions, and Damien Laouteouet and Jean-Christophe Perez for generating and performing initial experiments with the artificial spectrin mutants. This work was supported by grants from the Agence Nationale de la Recherche to AD (ANR-2019 TRIOTISM) and to CF (ANR-20-CE16-0019) and from The Fondation pour la Recherche Médicale (program Equipes FRM2016, DEQ20160334942) to AD. MaxB and MarB are recipients of a PhD fellowship from the Ministère de l’Enseignement Supérieur et de la Recherche (MESR). DB is generously supported by a National Institute for Health and Care Research research professorship RP-2016-07-011. We acknowledge the imaging facility MRI, and in particular Volker Bäcker, part of the national France-BioImaging infrastructure supported by the French National Research Agency (ANR-10-INBS-04, “Investments for the future”). Publisher Copyright: © 2023, The Author(s), under exclusive licence to Springer Nature Limited.

Identifiers

Local EPrints ID: 474135
URI: http://eprints.soton.ac.uk/id/eprint/474135
ISSN: 1359-4184
PURE UUID: 5aaaca0e-ab33-4573-8499-0b49a0c811fb
ORCID for Diana Baralle: ORCID iD orcid.org/0000-0003-3217-4833

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Date deposited: 14 Feb 2023 17:37
Last modified: 13 Aug 2024 04:01

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Contributors

Author: Maxime Bonnet
Author: Fiona Roche
Author: Christine Fagotto-Kaufmann
Author: Gabriella Gazdagh
Author: Iona Truong
Author: Franck Comunale
Author: Sonia Barbosa
Author: Marion Bonhomme
Author: Nicholas Nafati
Author: David Hunt
Author: Monserrat Pons Rodriguez
Author: Ayeshah Chaudhry
Author: Deborah Shears
Author: Marcos Madruga
Author: Fleur Vansenne
Author: Aurore Curie
Author: Andry V Kajava
Author: Diana Baralle ORCID iD
Author: Coralie Fassier
Author: Anne Debant
Author: Susanne Schmidt

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