TY - JOUR
T1 - TBK1 phosphorylates mutant Huntingtin and suppresses its aggregation and toxicity in Huntington's disease models
AU - Hegde, Ramanath Narayana
AU - Chiki, Anass
AU - Petricca, Lara
AU - Martufi, Paola
AU - Arbez, Nicolas
AU - Mouchiroud, Laurent
AU - Auwerx, Johan
AU - Landles, Christian
AU - Bates, Gillian P.
AU - Singh-Bains, Malvindar K.
AU - Dragunow, Mike
AU - Curtis, Maurice A.
AU - Faull, Richard L.M.
AU - Ross, Christopher A.
AU - Caricasole, Andrea
AU - Lashuel, Hilal A.
N1 - Funding Information:
This work was supported by funding from CHDI and EPFL and NINDS (R01 NS086452 and R21 NS083365). We are grateful to Anne‐Laure Mahul‐Mellier, Galina Limorenko, Johannes Burtscher, Niran Maharjan, Senthil Kumar Thangaraj, and Somanath Jagannath, EPFL, for critical review of the manuscript. We thank Rajasekhar Kolla for preparing the graphical abstract. We are grateful to staff at the Bio‐imaging Core Facility (BioP, EPFL) for their technical support. We thank Driss Boudeffa, EPFL, for the production of lentiviral vectors; Jonathan Ricci, EPFL, for assisting kinase validation; and Elena Gasparotto and Lorène Aeschbach, EPFL, for assisting with neuronal cell culture experiments. We thank Naveed Ziari and Kevin S. Hof, EPFL‐LISP‐IBI‐SV, for assisting with experiments in Fig 5 B, C, H, I and Appendix Fig S2 and initial help with the setting up of model. We thank the proteomics core facility at EPFL's School of Life Sciences and Functional Genomics Center Zurich (FGCZ), ETH Zurich, for their support with the mass spectrometry analysis. We are grateful to Prof. Elise A. Kikis, The University of the South, Sewanee, Tennessee, for the kind gift of HD model and Prof. Shizuo Akira, WPI Immunology Frontier Research Center, Osaka University, Japan, for the kind gift of mouse model used in this study. in vitro C. elegans C. elegans Tbk1 −/−
Funding Information:
This work was supported by funding from CHDI and EPFL and NINDS (R01 NS086452 and R21 NS083365). We are grateful to Anne-Laure Mahul-Mellier, Galina Limorenko, Johannes Burtscher, Niran Maharjan, Senthil Kumar Thangaraj, and Somanath Jagannath, EPFL, for critical review of the manuscript. We thank Rajasekhar Kolla for preparing the graphical abstract. We are grateful to staff at the Bio-imaging Core Facility (BioP, EPFL) for their technical support. We thank Driss Boudeffa, EPFL, for the production of lentiviral vectors; Jonathan Ricci, EPFL, for assisting in?vitro kinase validation; and Elena Gasparotto and Lor?ne Aeschbach, EPFL, for assisting with neuronal cell culture experiments. We thank Naveed Ziari and Kevin S. Hof, EPFL-LISP-IBI-SV, for assisting with experiments in Fig?5B, C, H, I and Appendix?Fig S2 and initial help with the setting up of C.?elegans model. We thank the proteomics core facility at EPFL's School of Life Sciences and Functional Genomics Center Zurich (FGCZ), ETH Zurich, for their support with the mass spectrometry analysis. We are grateful to Prof. Elise A. Kikis, The University of the South, Sewanee, Tennessee, for the kind gift of HD C.?elegans model and Prof. Shizuo Akira, WPI Immunology Frontier Research Center, Osaka University, Japan, for the kind gift of Tbk1?/? mouse model used in this study.
Publisher Copyright:
© 2020 The Authors. Published under the terms of the CC BY NC ND 4.0 license
PY - 2020/9/1
Y1 - 2020/9/1
N2 - Phosphorylation of the N-terminal domain of the huntingtin (HTT) protein has emerged as an important regulator of its localization, structure, aggregation, clearance and toxicity. However, validation of the effect of bona fide phosphorylation in vivo and assessing the therapeutic potential of targeting phosphorylation for the treatment of Huntington's disease (HD) require the identification of the enzymes that regulate HTT phosphorylation. Herein, we report the discovery and validation of a kinase, TANK-binding kinase 1 (TBK1), that efficiently phosphorylates full-length and N-terminal HTT fragments in vitro (at S13/S16), in cells (at S13) and in vivo. TBK1 expression in HD models (cells, primary neurons, and Caenorhabditis elegans) increases mutant HTT exon 1 phosphorylation and reduces its aggregation and cytotoxicity. We demonstrate that the TBK1-mediated neuroprotective effects are due to phosphorylation-dependent inhibition of mutant HTT exon 1 aggregation and an increase in autophagic clearance of mutant HTT. These findings suggest that upregulation and/or activation of TBK1 represents a viable strategy for the treatment of HD by simultaneously lowering mutant HTT levels and blocking its aggregation.
AB - Phosphorylation of the N-terminal domain of the huntingtin (HTT) protein has emerged as an important regulator of its localization, structure, aggregation, clearance and toxicity. However, validation of the effect of bona fide phosphorylation in vivo and assessing the therapeutic potential of targeting phosphorylation for the treatment of Huntington's disease (HD) require the identification of the enzymes that regulate HTT phosphorylation. Herein, we report the discovery and validation of a kinase, TANK-binding kinase 1 (TBK1), that efficiently phosphorylates full-length and N-terminal HTT fragments in vitro (at S13/S16), in cells (at S13) and in vivo. TBK1 expression in HD models (cells, primary neurons, and Caenorhabditis elegans) increases mutant HTT exon 1 phosphorylation and reduces its aggregation and cytotoxicity. We demonstrate that the TBK1-mediated neuroprotective effects are due to phosphorylation-dependent inhibition of mutant HTT exon 1 aggregation and an increase in autophagic clearance of mutant HTT. These findings suggest that upregulation and/or activation of TBK1 represents a viable strategy for the treatment of HD by simultaneously lowering mutant HTT levels and blocking its aggregation.
KW - Huntington's disease
KW - TBK1
KW - autophagy
KW - huntingtin phosphorylation
KW - reducing aggregation
UR - http://www.scopus.com/inward/record.url?scp=85088928225&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85088928225&partnerID=8YFLogxK
U2 - 10.15252/embj.2020104671
DO - 10.15252/embj.2020104671
M3 - Article
C2 - 32757223
AN - SCOPUS:85088928225
SN - 0261-4189
VL - 39
JO - EMBO Journal
JF - EMBO Journal
IS - 17
M1 - e104671
ER -