Excessive proteostasis contributes to pathology in fragile X syndrome

Susana R. Louros; Sang S. Seo; Beatriz Maio; Cristina Martinez-Gonzalez; Miguel A. Gonzalez-Lozano; Melania Muscas; Nick C. Verity; Jimi C. Wills; Ka Wan Li; Matthew F. Nolan; Emily K. Osterweil

doi:10.1016/j.neuron.2022.11.012

Excessive proteostasis contributes to pathology in fragile X syndrome

Susana R. Louros, Sang S. Seo, Beatriz Maio, Cristina Martinez-Gonzalez, Miguel A. Gonzalez-Lozano, Melania Muscas, Nick C. Verity, Jimi C. Wills, Ka Wan Li, Matthew F. Nolan, Emily K. Osterweil^*

^*Corresponding author for this work

Research output: Contribution to Journal › Article › Academic › peer-review

Abstract

In fragile X syndrome (FX), the leading monogenic cause of autism, excessive neuronal protein synthesis is a core pathophysiology; however, an overall increase in protein expression is not observed. Here, we tested whether excessive protein synthesis drives a compensatory rise in protein degradation that is protective for FX mouse model (Fmr1^−/y) neurons. Surprisingly, although we find a significant increase in protein degradation through ubiquitin proteasome system (UPS), this contributes to pathological changes. Normalizing proteasome activity with bortezomib corrects excessive hippocampal protein synthesis and hyperactivation of neurons in the inferior colliculus (IC) in response to auditory stimulation. Moreover, systemic administration of bortezomib significantly reduces the incidence and severity of audiogenic seizures (AGS) in the Fmr1^−/y mouse, as does genetic reduction of proteasome, specifically in the IC. Together, these results identify excessive activation of the UPS pathway in Fmr1^−/y neurons as a contributor to multiple phenotypes that can be targeted for therapeutic intervention.

Original language	English
Pages (from-to)	508-525.e7
Number of pages	26
Journal	Neuron
Volume	111
Issue number	4
Early online date	9 Dec 2022
DOIs	https://doi.org/10.1016/j.neuron.2022.11.012
Publication status	Published - 15 Feb 2023

Bibliographical note

Funding Information:
Special thanks to Steph Barnes, Caoimhe Kirby, and Laura Kaminioti-Dumont for help with biochemistry and AGS experiments. The authors are grateful for support from the Wellcome Trust (Sir Henry Dale fellowship 104116/Z/14/Z; Senior Research Fellowship 219556/Z/19/Z), Medical Research Council (MR/S026312/1), Simons Initiative for the Developing Brain (SIDB), and ONO Pharmaceuticals. We thank the IMPACT facility at the University of Edinburgh for imaging resources. E.K.O. S.R.L. and S.S.S. conceptualized the study and prepared the manuscript. M.F.N. provided essential feedback on multiple aspects of the study and was the major contributor to the conception, design, and analyses of c-Fos+ imaging experiments and oversight of data acquisition by S.R.L. and C.M.-G. S.R.L. designed, performed, and analyzed hippocampal slice experiments and all biochemistry experiments, including the preparation of samples for proteomic analysis. S.R.L. designed, performed, and analyzed AGS experiments with assistance from M.M. and B.M. S.S.S. performed bioinformatics analyses of TRAP-seq datasets and proteomics datasets from hippocampus and IC. M.A.G.-L. and K.W.L. performed MS analysis of hippocampal samples. J.C.W. performed MS analysis of IC samples. S.S.S. and N.C.V. performed viral injections, TRAP experiments, and qPCR with assistance from B.M. S.R.L. and B.M. performed c-Fos immunostaining and imaging experiments with assistance from C.M.-G. S.R.L. and C.M.-G. performed c-Fos+ neuron quantification. All authors contributed to the review and editing of the manuscript, with major inputs from S.R.L. S.S.S. and M.F.N. The authors declare no competing interests. One or more of the authors of this paper self-identifies as an underrepresented ethnic minority in their field of research or within their geographical location. One or more of the authors of this paper self-identifies as a gender minority in their field of research. One or more of the authors of this paper self-identifies as a member of the LGBTQIA+ community. One or more of the authors of this paper self-identifies as living with a disability. While citing references scientifically relevant for this work, we also actively worked to promote gender balance in our reference list.

Funding Information:
Special thanks to Steph Barnes, Caoimhe Kirby, and Laura Kaminioti-Dumont for help with biochemistry and AGS experiments. The authors are grateful for support from the Wellcome Trust (Sir Henry Dale fellowship 104116/Z/14/Z ; Senior Research Fellowship 219556/Z/19/Z ), Medical Research Council ( MR/S026312/1 ), Simons Initiative for the Developing Brain (SIDB), and ONO Pharmaceuticals . We thank the IMPACT facility at the University of Edinburgh for imaging resources.

Publisher Copyright:
© 2022 The Author(s)

Keywords

AGS
bortezomib
FMR1
fragile X
inferior colliculus
proteasome
proteostasis
PSMB5
seizures
UPS

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.neuron.2022.11.012

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title = "Excessive proteostasis contributes to pathology in fragile X syndrome",

abstract = "In fragile X syndrome (FX), the leading monogenic cause of autism, excessive neuronal protein synthesis is a core pathophysiology; however, an overall increase in protein expression is not observed. Here, we tested whether excessive protein synthesis drives a compensatory rise in protein degradation that is protective for FX mouse model (Fmr1−/y) neurons. Surprisingly, although we find a significant increase in protein degradation through ubiquitin proteasome system (UPS), this contributes to pathological changes. Normalizing proteasome activity with bortezomib corrects excessive hippocampal protein synthesis and hyperactivation of neurons in the inferior colliculus (IC) in response to auditory stimulation. Moreover, systemic administration of bortezomib significantly reduces the incidence and severity of audiogenic seizures (AGS) in the Fmr1−/y mouse, as does genetic reduction of proteasome, specifically in the IC. Together, these results identify excessive activation of the UPS pathway in Fmr1−/y neurons as a contributor to multiple phenotypes that can be targeted for therapeutic intervention.",

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note = "Funding Information: Special thanks to Steph Barnes, Caoimhe Kirby, and Laura Kaminioti-Dumont for help with biochemistry and AGS experiments. The authors are grateful for support from the Wellcome Trust (Sir Henry Dale fellowship 104116/Z/14/Z; Senior Research Fellowship 219556/Z/19/Z), Medical Research Council (MR/S026312/1), Simons Initiative for the Developing Brain (SIDB), and ONO Pharmaceuticals. We thank the IMPACT facility at the University of Edinburgh for imaging resources. E.K.O. S.R.L. and S.S.S. conceptualized the study and prepared the manuscript. M.F.N. provided essential feedback on multiple aspects of the study and was the major contributor to the conception, design, and analyses of c-Fos+ imaging experiments and oversight of data acquisition by S.R.L. and C.M.-G. S.R.L. designed, performed, and analyzed hippocampal slice experiments and all biochemistry experiments, including the preparation of samples for proteomic analysis. S.R.L. designed, performed, and analyzed AGS experiments with assistance from M.M. and B.M. S.S.S. performed bioinformatics analyses of TRAP-seq datasets and proteomics datasets from hippocampus and IC. M.A.G.-L. and K.W.L. performed MS analysis of hippocampal samples. J.C.W. performed MS analysis of IC samples. S.S.S. and N.C.V. performed viral injections, TRAP experiments, and qPCR with assistance from B.M. S.R.L. and B.M. performed c-Fos immunostaining and imaging experiments with assistance from C.M.-G. S.R.L. and C.M.-G. performed c-Fos+ neuron quantification. All authors contributed to the review and editing of the manuscript, with major inputs from S.R.L. S.S.S. and M.F.N. The authors declare no competing interests. One or more of the authors of this paper self-identifies as an underrepresented ethnic minority in their field of research or within their geographical location. One or more of the authors of this paper self-identifies as a gender minority in their field of research. One or more of the authors of this paper self-identifies as a member of the LGBTQIA+ community. One or more of the authors of this paper self-identifies as living with a disability. While citing references scientifically relevant for this work, we also actively worked to promote gender balance in our reference list. Funding Information: Special thanks to Steph Barnes, Caoimhe Kirby, and Laura Kaminioti-Dumont for help with biochemistry and AGS experiments. The authors are grateful for support from the Wellcome Trust (Sir Henry Dale fellowship 104116/Z/14/Z ; Senior Research Fellowship 219556/Z/19/Z ), Medical Research Council ( MR/S026312/1 ), Simons Initiative for the Developing Brain (SIDB), and ONO Pharmaceuticals . We thank the IMPACT facility at the University of Edinburgh for imaging resources. Publisher Copyright: {\textcopyright} 2022 The Author(s)",

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AU - Seo, Sang S.

AU - Maio, Beatriz

AU - Martinez-Gonzalez, Cristina

AU - Gonzalez-Lozano, Miguel A.

AU - Muscas, Melania

AU - Verity, Nick C.

AU - Wills, Jimi C.

AU - Li, Ka Wan

AU - Nolan, Matthew F.

AU - Osterweil, Emily K.

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PY - 2023/2/15

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N2 - In fragile X syndrome (FX), the leading monogenic cause of autism, excessive neuronal protein synthesis is a core pathophysiology; however, an overall increase in protein expression is not observed. Here, we tested whether excessive protein synthesis drives a compensatory rise in protein degradation that is protective for FX mouse model (Fmr1−/y) neurons. Surprisingly, although we find a significant increase in protein degradation through ubiquitin proteasome system (UPS), this contributes to pathological changes. Normalizing proteasome activity with bortezomib corrects excessive hippocampal protein synthesis and hyperactivation of neurons in the inferior colliculus (IC) in response to auditory stimulation. Moreover, systemic administration of bortezomib significantly reduces the incidence and severity of audiogenic seizures (AGS) in the Fmr1−/y mouse, as does genetic reduction of proteasome, specifically in the IC. Together, these results identify excessive activation of the UPS pathway in Fmr1−/y neurons as a contributor to multiple phenotypes that can be targeted for therapeutic intervention.

AB - In fragile X syndrome (FX), the leading monogenic cause of autism, excessive neuronal protein synthesis is a core pathophysiology; however, an overall increase in protein expression is not observed. Here, we tested whether excessive protein synthesis drives a compensatory rise in protein degradation that is protective for FX mouse model (Fmr1−/y) neurons. Surprisingly, although we find a significant increase in protein degradation through ubiquitin proteasome system (UPS), this contributes to pathological changes. Normalizing proteasome activity with bortezomib corrects excessive hippocampal protein synthesis and hyperactivation of neurons in the inferior colliculus (IC) in response to auditory stimulation. Moreover, systemic administration of bortezomib significantly reduces the incidence and severity of audiogenic seizures (AGS) in the Fmr1−/y mouse, as does genetic reduction of proteasome, specifically in the IC. Together, these results identify excessive activation of the UPS pathway in Fmr1−/y neurons as a contributor to multiple phenotypes that can be targeted for therapeutic intervention.

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KW - seizures

KW - UPS

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SN - 0896-6273

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JO - Neuron

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Excessive proteostasis contributes to pathology in fragile X syndrome

Abstract

Bibliographical note

Keywords

UN SDGs

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