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EPHA4 is a disease modifier of amyotrophic lateral sclerosis in animal models and in humans

Abstract

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease affecting motor neurons. Disease onset and progression are variable, with survival ranging from months to decades. Factors underlying this variability may represent targets for therapeutic intervention. Here, we have screened a zebrafish model of ALS and identified Epha4, a receptor in the ephrin axonal repellent system, as a modifier of the disease phenotype in fish, rodents and humans. Genetic as well as pharmacological inhibition of Epha4 signaling rescues the mutant SOD1 phenotype in zebrafish and increases survival in mouse and rat models of ALS. Motor neurons that are most vulnerable to degeneration in ALS express higher levels of Epha4, and neuromuscular re-innervation by axotomized motor neurons is inhibited by the presence of Epha4. In humans with ALS, EPHA4 expression inversely correlates with disease onset and survival, and loss-of-function mutations in EPHA4 are associated with long survival. Furthermore, we found that knockdown of Epha4 also rescues the axonopathy induced by expression of mutant TAR DNA-binding protein 43 (TDP-43), another protein causing familial ALS, and the axonopathy induced by knockdown of survival of motor neuron 1, a model for spinomuscular atrophy. This suggests that Epha4 generically modulates the vulnerability of (motor) neurons to axonal degeneration and may represent a new target for therapeutic intervention.

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Figure 1: Epha4 knockdown rescues a mutant SOD1–induced motor axonopathy.
Figure 2: Genetic knockdown and pharmacological inhibition of Epha4 attenuates disease.
Figure 3: Vulnerable motor neurons in ALS have higher expression of Epha4.
Figure 4: EPHA4 attenuates disease progression in humans with ALS and in motor axonopathies induced by overexpression of mutant TDP-43 and knockdown of Smn1.

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Acknowledgements

This work has been supported by grants from the Flanders Institute for Biotechnology (VIB), the University of Leuven (GOA 11/014), Life Sciences Research Partners, Research Foundation Flanders (FWO-Vlaanderen; G.0695.10), the Interuniversity Attraction Poles program P7/16 of the Belgian Federal Science Policy Office, the Robert Packard Center for ALS Research, the Association Belge contre les Maladies Musculaires, the ALS League Belgium and the Thierry Latran Foundation. The research leading to these results has received funding from the European Community's Health Seventh Framework Programme (FP7/2007-2013) under grant agreement number 259867. W.R. is supported by the E. von Behring Chair for Neuromuscular and Neurodegenerative Disorders at the University of Leuven. A.V.H. is supported by the Agency for Innovation by Science and Technology in Flanders (IWT-Vlaanderen). R.L., P.V.D., V.T. and B.D. hold a clinical investigatorship of FWO-Vlaanderen. R.L. is supported by Research Fund KU Leuven. P.G.-P. is supported by the Alfonso Martin Escudero Foundation. A.A.-C. receives salary support from the UK National Institute for Health Research (NIHR) Dementia Biomedical Research Unit at South London and Maudsley National Health Service (NHS) Foundation Trust and King's College London. The views expressed are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health. R.H.B. Jr. is supported by the US National Institute for Neurological Disease and Stroke (NINDS) (5RO1-NS050557-05) and NINDS American Recovery and Reinvestment Act Award RC2-NS070-342 and acknowledges generous support from the Angel Fund, the ALS Association, P2ALS, Project ALS, the Pierre L. de Bourgknecht ALS Research Foundation and the ALS Therapy Alliance.

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W.R. designed the study, analyzed data and wrote the manuscript. A.V.H. designed and performed experiments, analyzed data and wrote the manuscript. R.L. collected samples, analyzed data and designed experiments. P.V.D. and L.V.D.B. collected samples and designed experiments. L.S., M.T., P.G.-P., K.A.S., A.S.L., T.P., E.P., A.G. and P.W.v.V. performed experiments. P.M.A., A.A.-C., O.H., J.H.V., L.H.v.d.B., V.T., R.H.B. Jr. and B.D. collected samples and analyzed data. A.M.T. provided mice. All authors discussed the results and implications and commented on the manuscript at all stages.

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Correspondence to Wim Robberecht.

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Supplementary Tables 1–10 and Supplementary Figures 1–8 (PDF 1399 kb)

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Van Hoecke, A., Schoonaert, L., Lemmens, R. et al. EPHA4 is a disease modifier of amyotrophic lateral sclerosis in animal models and in humans. Nat Med 18, 1418–1422 (2012). https://doi.org/10.1038/nm.2901

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