Volume 44 Issue 2
Mar.  2023
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Wen-Yo Tu, Wentao Xu, Jianmin Zhang, Shuyuan Qi, Lei Bai, Chengyong Shen, Kejing Zhang. C9orf72 poly-GA proteins impair neuromuscular transmission. Zoological Research, 2023, 44(2): 331-340. doi: 10.24272/j.issn.2095-8137.2022.356
Citation: Wen-Yo Tu, Wentao Xu, Jianmin Zhang, Shuyuan Qi, Lei Bai, Chengyong Shen, Kejing Zhang. C9orf72 poly-GA proteins impair neuromuscular transmission. Zoological Research, 2023, 44(2): 331-340. doi: 10.24272/j.issn.2095-8137.2022.356

C9orf72 poly-GA proteins impair neuromuscular transmission

doi: 10.24272/j.issn.2095-8137.2022.356
Supplementary data to this article can be found online.
The authors declare that they have no competing interests.
K.Z. and C.S.: Conceptualization, supervision, resources, and writing. W.Y.T. and W.X.: Methodology, investigation, and writing. J.Z., S.Q., and L.B.: Investigation. All authors read and approved the final version of the manuscript.
#Authors contributed equally to this work
Funds:  This work was supported by the National Key Research and Development Program of China (2022YFF1000500 to K.Z. and 2021YFA1101100 to C.S.), Zhejiang Provincial Natural Science Foundation (LZ22C110002 to C.S.), and National Natural Science Foundation of China (32271031 to K.Z. and 82230038, 31871203, and 32071032 to C.S.)
More Information
  • Corresponding author: E-mail: cshen@zju.edu.cnkjzhang@zju.edu.cn
  • Received Date: 2022-12-14
  • Accepted Date: 2023-02-15
  • Published Online: 2023-02-17
  • Publish Date: 2023-03-18
  • Amyotrophic lateral sclerosis (ALS) is a devastating motoneuron disease, in which lower motoneurons lose control of skeletal muscles. Degeneration of neuromuscular junctions (NMJs) occurs at the initial stage of ALS. Dipeptide repeat proteins (DPRs) from G4C2 repeat-associated non-ATG (RAN) translation are known to cause C9orf72-associated ALS (C9-ALS). However, DPR inclusion burdens are weakly correlated with neurodegenerative areas in C9-ALS patients, indicating that DPRs may exert cell non-autonomous effects, in addition to the known intracellular pathological mechanisms. Here, we report that poly-GA, the most abundant form of DPR in C9-ALS, is released from cells. Local administration of poly-GA proteins in peripheral synaptic regions causes muscle weakness and impaired neuromuscular transmission in vivo. The NMJ structure cannot be maintained, as evidenced by the fragmentation of postsynaptic acetylcholine receptor (AChR) clusters and distortion of presynaptic nerve terminals. Mechanistic study demonstrated that extracellular poly-GA sequesters soluble Agrin ligands and inhibits Agrin-MuSK signaling. Our findings provide a novel cell non-autonomous mechanism by which poly-GA impairs NMJs in C9-ALS. Thus, targeting NMJs could be an early therapeutic intervention for C9-ALS.
  • Supplementary data to this article can be found online.
    The authors declare that they have no competing interests.
    K.Z. and C.S.: Conceptualization, supervision, resources, and writing. W.Y.T. and W.X.: Methodology, investigation, and writing. J.Z., S.Q., and L.B.: Investigation. All authors read and approved the final version of the manuscript.
    #Authors contributed equally to this work
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  • [1]
    Arimura S, Okada T, Tezuka T, et al. 2014. DOK7 gene therapy benefits mouse models of diseases characterized by defects in the neuromuscular junction. Science, 345(6203): 1505−1508. doi: 10.1126/science.1250744
    [2]
    Bai L, Tu WY, Xiao YT, et al. 2022. Motoneurons innervation determines the distinct gene expressions in multinucleated myofibers. Cell & Bioscience, 12(1): 140.
    [3]
    Bentzinger CF, Barzaghi P, Lin S, et al. 2005. Overexpression of mini-agrin in skeletal muscle increases muscle integrity and regenerative capacity in laminin-α2-deficient mice. The FASEB Journal, 19(8): 934−942. doi: 10.1096/fj.04-3376com
    [4]
    Bertrand A, Wen J, Rinaldi D, et al. 2018. Early cognitive, structural, and microstructural changes in presymptomatic C9orf72 Carriers younger than 40 years. JAMA Neurology, 75(2): 236−245. doi: 10.1001/jamaneurol.2017.4266
    [5]
    Bezakova G, Ruegg MA. 2003. New insights into the roles of agrin. Nature Reviews Molecular Cell Biology, 4(4): 295−309. doi: 10.1038/nrm1074
    [6]
    Brahic M, Bousset L, Bieri G, et al. 2016. Axonal transport and secretion of fibrillar forms of α-synuclein, Aβ42 peptide and HTTExon 1. Acta Neuropathologica, 131(4): 539−548. doi: 10.1007/s00401-016-1538-0
    [7]
    Burden SJ, Yumoto N, Zhang W. 2013. The role of MuSK in synapse formation and neuromuscular disease. Cold Spring Harbor Perspectives in Biology, 5(5): a009167.
    [8]
    Chand KK, Lee KM, Lee JD, et al. 2018. Defects in synaptic transmission at the neuromuscular junction precede motor deficits in a TDP-43Q331K transgenic mouse model of amyotrophic lateral sclerosis. The FASEB Journal, 32(5): 2676−2689. doi: 10.1096/fj.201700835R
    [9]
    Chang YJ, Jeng US, Chiang YL, et al. 2016. The glycine-alanine dipeptide repeat from C9orf72 hexanucleotide expansions forms toxic amyloids possessing cell-to-cell transmission properties. Journal of Biological Chemistry, 291(10): 4903−4911. doi: 10.1074/jbc.M115.694273
    [10]
    Chen AZ, Bai L, Zhong KK, et al. 2020. APC2CDH1 negatively regulates agrin signaling by promoting the ubiquitination and proteolytic degradation of DOK7. The FASEB Journal, 34(9): 12009−12023. doi: 10.1096/fj.202000485R
    [11]
    Cleary JD, Ranum LPW. 2017. New developments in RAN translation: insights from multiple diseases. Current Opinion in Genetics & Development, 44: 125−134.
    [12]
    DeJesus-Hernandez M, Mackenzie IR, Boeve BF, et al. 2011. Expanded GGGGCC hexanucleotide repeat in noncoding region of C9ORF72 causes chromosome 9p-linked FTD and ALS. Neuron, 72(2): 245−256. doi: 10.1016/j.neuron.2011.09.011
    [13]
    Flores BN, Dulchavsky ME, Krans A, et al. 2016. Distinct C9orf72-associated dipeptide repeat structures correlate with neuronal toxicity. PLoS One, 11(10): e0165084. doi: 10.1371/journal.pone.0165084
    [14]
    Freibaum BD, Lu YB, Lopez-Gonzalez R, et al. 2015. GGGGCC repeat expansion in C9orf72 compromises nucleocytoplasmic transport. Nature, 525(7567): 129−133. doi: 10.1038/nature14974
    [15]
    Gao FB, Almeida S, Lopez-Gonzalez R. 2017a. Dysregulated molecular pathways in amyotrophic lateral sclerosis-frontotemporal dementia spectrum disorder. The EMBO Journal, 36(20): 2931−2950. doi: 10.15252/embj.201797568
    [16]
    Gao FB, Richter JD, Cleveland DW. 2017b. Rethinking unconventional translation in neurodegeneration. Cell, 171(5): 994−1000. doi: 10.1016/j.cell.2017.10.042
    [17]
    Geevasinga N, Menon P, Özdinler PH, et al. 2016. Pathophysiological and diagnostic implications of cortical dysfunction in ALS. Nature Reviews Neurology, 12(11): 651−661. doi: 10.1038/nrneurol.2016.140
    [18]
    Gendron TF, Chew J, Stankowski JN, et al. 2017. Poly(GP) proteins are a useful pharmacodynamic marker for C9ORF72-associated amyotrophic lateral sclerosis. Science Translational Medicine, 9(383): eaai7866. doi: 10.1126/scitranslmed.aai7866
    [19]
    Gendron TF, van Blitterswijk M, Bieniek KF, et al. 2015. Cerebellar c9RAN proteins associate with clinical and neuropathological characteristics of C9ORF72 repeat expansion carriers. Acta Neuropathologica, 130(4): 559−573. doi: 10.1007/s00401-015-1474-4
    [20]
    Gilhus NE, Skeie GO, Romi F, et al. 2016. Myasthenia gravis - autoantibody characteristics and their implications for therapy. Nature Reviews Neurology, 12(5): 259−268. doi: 10.1038/nrneurol.2016.44
    [21]
    Haeusler AR, Donnelly CJ, Rothstein JD. 2016. The expanding biology of the C9orf72 nucleotide repeat expansion in neurodegenerative disease. Nature Reviews Neuroscience, 17(6): 383−395. doi: 10.1038/nrn.2016.38
    [22]
    Herranz-Martin S, Chandran J, Lewis K, et al. 2017. Viral delivery of C9orf72 hexanucleotide repeat expansions in mice leads to repeat-length-dependent neuropathology and behavioural deficits. Disease Models & Mechanisms, 10(7): 859−868.
    [23]
    Hettwer S, Lin S, Kucsera S, et al. 2014. Injection of a soluble fragment of neural agrin (NT-1654) considerably improves the muscle pathology caused by the disassembly of the neuromuscular junction. PLoS One, 9(2): e88739. doi: 10.1371/journal.pone.0088739
    [24]
    Jensen BK, Schuldi MH, McAvoy K, et al. 2020. Synaptic dysfunction induced by glycine-alanine dipeptides in C9orf72-ALS/FTD is rescued by SV2 replenishment. EMBO Molecular Medicine, 12(5): e10722.
    [25]
    Kim N, Stiegler AL, Cameron TO, et al. 2008. Lrp4 is a receptor for Agrin and forms a complex with MuSK. Cell, 135(2): 334−342. doi: 10.1016/j.cell.2008.10.002
    [26]
    Krishnan G, Raitcheva D, Bartlett D, et al. 2022. Poly(GR) and poly(GA) in cerebrospinal fluid as potential biomarkers for C9ORF72-ALS/FTD. Nature Communications, 13(1): 2799. doi: 10.1038/s41467-022-30387-4
    [27]
    Lee YB, Baskaran P, Gomez-Deza J, et al. 2017. C9orf72 poly GA RAN-translated protein plays a key role in amyotrophic lateral sclerosis via aggregation and toxicity. Human Molecular Genetics, 26(24): 4765−4777. doi: 10.1093/hmg/ddx350
    [28]
    Li L, Xiong WC, Mei L. 2018. Neuromuscular junction formation, aging, and disorders. Annual Review of Physiology, 80: 159−188. doi: 10.1146/annurev-physiol-022516-034255
    [29]
    Li WY, Wang Y, Zhai FG, et al. 2017. AAV-KLF7 promotes descending propriospinal neuron axonal plasticity after spinal cord injury. Neural Plasticity, 2017: 1621629.
    [30]
    Liu YJ, Pattamatta A, Zu T, et al. 2016. C9orf72 BAC mouse model with motor deficits and neurodegenerative features of ALS/FTD. Neuron, 90(3): 521−534. doi: 10.1016/j.neuron.2016.04.005
    [31]
    Mackenzie IR, Arzberger T, Kremmer E, et al. 2013. Dipeptide repeat protein pathology in C9ORF72 mutation cases: clinico-pathological correlations. Acta Neuropathologica, 126(6): 859−879. doi: 10.1007/s00401-013-1181-y
    [32]
    May S, Hornburg D, Schludi MH, et al. 2014. C9orf72 FTLD/ALS-associated Gly-Ala dipeptide repeat proteins cause neuronal toxicity and Unc119 sequestration. Acta Neuropathologica, 128(4): 485−503. doi: 10.1007/s00401-014-1329-4
    [33]
    Miyoshi S, Tezuka T, Arimura S, et al. 2017. DOK7 gene therapy enhances motor activity and life span in ALS model mice. EMBO Molecular Medicine, 9(7): 880−889. doi: 10.15252/emmm.201607298
    [34]
    Moloney EB, De Winter F, Verhaagen J. 2014. ALS as a distal axonopathy: molecular mechanisms affecting neuromuscular junction stability in the presymptomatic stages of the disease. Frontiers in Neuroscience, 8: 252.
    [35]
    Moretto E, Stuart S, Surana S, et al. 2022. The role of extracellular matrix components in the spreading of pathological protein aggregates. Frontiers in Cellular Neuroscience, 16: 844211. doi: 10.3389/fncel.2022.844211
    [36]
    Mori K, Weng SM, Arzberger T, et al. 2013. The C9orf72 GGGGCC repeat is translated into aggregating dipeptide-repeat proteins in FTLD/ALS. Science, 339(6125): 1335−1338. doi: 10.1126/science.1232927
    [37]
    Nguyen L, Montrasio F, Pattamatta A, et al. 2020. Antibody therapy targeting RAN proteins rescues C9 ALS/FTD phenotypes in C9orf72 mouse model. Neuron, 105(4): 645−662.e11. doi: 10.1016/j.neuron.2019.11.007
    [38]
    Ohno K, Ohkawara B, Ito M. 2017. Agrin-LRP4-MuSK signaling as a therapeutic target for myasthenia gravis and other neuromuscular disorders. Expert Opinion on Therapeutic Targets, 21(10): 949−958. doi: 10.1080/14728222.2017.1369960
    [39]
    O'Rourke JG, Bogdanik L, Yáñez A, et al. 2016. C9orf72 is required for proper macrophage and microglial function in mice. Science, 351(6279): 1324−1329. doi: 10.1126/science.aaf1064
    [40]
    Pérez-García MJ, Burden SJ. 2012. Increasing MuSK activity delays denervation and improves motor function in ALS mice. Cell Reports, 2(3): 497−502. doi: 10.1016/j.celrep.2012.08.004
    [41]
    Renton AE, Majounie E, Waite A, et al. 2011. A hexanucleotide repeat expansion in C9ORF72 is the cause of chromosome 9p21-linked ALS-FTD. Neuron, 72(2): 257−268. doi: 10.1016/j.neuron.2011.09.010
    [42]
    Rivner MH, Liu SY, Quarles B, et al. 2017. Agrin and low-density lipoprotein-related receptor protein 4 antibodies in amyotrophic lateral sclerosis patients. Muscle & Nerve, 55(3): 430−432.
    [43]
    Rodriguez Cruz PM, Palace J, Beeson D. 2014. Congenital myasthenic syndromes and the neuromuscular junction. Current Opinion in Neurology, 27(5): 566−575. doi: 10.1097/WCO.0000000000000134
    [44]
    Saberi S, Stauffer JE, Jiang J, et al. 2018. Sense-encoded poly-GR dipeptide repeat proteins correlate to neurodegeneration and uniquely co-localize with TDP-43 in dendrites of repeat-expanded C9orf72 amyotrophic lateral sclerosis. Acta Neuropathologica, 135(3): 459−474. doi: 10.1007/s00401-017-1793-8
    [45]
    Schludi MH, Becker L, Garrett L, et al. 2017. Spinal poly-GA inclusions in a C9orf72 mouse model trigger motor deficits and inflammation without neuron loss. Acta Neuropathologica, 134(2): 241−254. doi: 10.1007/s00401-017-1711-0
    [46]
    Schludi MH, May S, Grässer FA, et al. 2015. Distribution of dipeptide repeat proteins in cellular models and C9orf72 mutation cases suggests link to transcriptional silencing. Acta Neuropathologica, 130(4): 537−555. doi: 10.1007/s00401-015-1450-z
    [47]
    Shen CY, Chen YF, Liu HQ, et al. 2008. Hydrogen peroxide promotes Aβ production through JNK-dependent activation of γ-secretase. Journal of Biological Chemistry, 283(25): 17721−17730. doi: 10.1074/jbc.M800013200
    [48]
    Shen CY, Li L, Zhao K, et al. 2018. Motoneuron Wnts regulate neuromuscular junction development. eLife, 7: e34625. doi: 10.7554/eLife.34625
    [49]
    Shen CY, Lu YS, Zhang B, et al. 2013. Antibodies against low-density lipoprotein receptor-related protein 4 induce myasthenia gravis. The Journal of Clinical Investigation, 123(12): 5190−5202. doi: 10.1172/JCI66039
    [50]
    Shen CY, Xiong WC, Mei L. 2015. LRP4 in neuromuscular junction and bone development and diseases. Bone, 80: 101−108. doi: 10.1016/j.bone.2015.05.012
    [51]
    Taylor JP, Brown RH Jr, Cleveland DW. 2016. Decoding ALS: from genes to mechanism. Nature, 539(7628): 197−206. doi: 10.1038/nature20413
    [52]
    Tu WY, Xu WT, Zhang KJ, et al. 2021. Whole-mount staining of neuromuscular junctions in adult mouse diaphragms with a sandwich-like apparatus. Journal of Neuroscience Methods, 350: 109016. doi: 10.1016/j.jneumeth.2020.109016
    [53]
    Tzartos JS, Zisimopoulou P, Rentzos M, et al. 2014. LRP4 antibodies in serum and CSF from amyotrophic lateral sclerosis patients. Annals of Clinical and Translational Neurology, 1(2): 80−87. doi: 10.1002/acn3.26
    [54]
    Westergard T, Jensen BK, Wen XM, et al. 2016. Cell-to-cell transmission of dipeptide repeat proteins linked to C9orf72-ALS/FTD. Cell Reports, 17(3): 645−652. doi: 10.1016/j.celrep.2016.09.032
    [55]
    Wu HT, Xiong WC, Mei L. 2010. To build a synapse: signaling pathways in neuromuscular junction assembly. Development, 137(7): 1017−1033. doi: 10.1242/dev.038711
    [56]
    Xiao YT, Zhang JM, Shu XQ, et al. 2020. Loss of mitochondrial protein CHCHD10 in skeletal muscle causes neuromuscular junction impairment. Human Molecular Genetics, 29(11): 1784−1796. doi: 10.1093/hmg/ddz154
    [57]
    Yamakawa M, Ito D, Honda T,et al. 2015. Characterization of the dipeptide repeat protein in the molecular pathogenesis of c9FTD/ALS. Human Molecular Genetics, 24(6): 1630−1645. doi: 10.1093/hmg/ddu576
    [58]
    Zhang B, Luo SW, Wang Q, et al. 2008. LRP4 serves as a coreceptor of agrin. Neuron, 60(2): 285−297. doi: 10.1016/j.neuron.2008.10.006
    [59]
    Zhang B, Shen CY, Bealmear B, et al. 2014. Autoantibodies to agrin in myasthenia gravis patients. PLoS One, 9(3): e91816. doi: 10.1371/journal.pone.0091816
    [60]
    Zhang K, Donnelly CJ, Haeusler AR, et al. 2015. The C9orf72 repeat expansion disrupts nucleocytoplasmic transport. Nature, 525(7567): 56−61. doi: 10.1038/nature14973
    [61]
    Zhang KJ, Wang AL, Zhong KK, et al. 2021. UBQLN2-HSP70 axis reduces poly-Gly-Ala aggregates and alleviates behavioral defects in the C9ORF72 animal model. Neuron, 109(12): 1949−1962.e6. doi: 10.1016/j.neuron.2021.04.023
    [62]
    Zhang YJ, Gendron TF, Grima JC, et al. 2016. C9ORF72 poly(GA) aggregates sequester and impair HR23 and nucleocytoplasmic transport proteins. Nature Neuroscience, 19(5): 668−677. doi: 10.1038/nn.4272
    [63]
    Zhao K, Shen CY, Lu YS, et al. 2017. Muscle yap is a regulator of neuromuscular junction formation and regeneration. Journal of Neuroscience, 37(13): 3465−3477. doi: 10.1523/JNEUROSCI.2934-16.2017
    [64]
    Zhou J, Li J, Rosenbaum DM, et al. 2017a. The prolyl 4-hydroxylase inhibitor GSK360A decreases post-stroke brain injury and sensory, motor, and cognitive behavioral deficits. PLoS One, 12(9): e0184049. doi: 10.1371/journal.pone.0184049
    [65]
    Zhou QH, Lehmer C, Michaelsen M, et al. 2017b. Antibodies inhibit transmission and aggregation of C9orf72 poly-GA dipeptide repeat proteins. EMBO Molecular Medicine, 9(5): 687−702. doi: 10.15252/emmm.201607054
    [66]
    Zu T, Gibbens B, Doty NS, et al. 2011. Non-ATG-initiated translation directed by microsatellite expansions. Proceedings of the National Academy of Sciences of the United States of America, 108(1): 260−265.
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