Top

Published in:

Open Access 01-12-2021 | Multiple Sclerosis | Research

Smek1 deficiency exacerbates experimental autoimmune encephalomyelitis by activating proinflammatory microglia and suppressing the IDO1-AhR pathway

Authors: Ruo-Nan Duan, Chun-Lin Yang, Tong Du, Ai Liu, An-Ran Wang, Wen-Jie Sun, Xi Li, Jiang-Xia Li, Chuan-Zhu Yan, Qi-Ji Liu

Published in: Journal of Neuroinflammation | Issue 1/2021

Abstract

Background

Experimental autoimmune encephalomyelitis (EAE) is an animal disease model of multiple sclerosis (MS) that involves the immune system and central nervous system (CNS). However, it is unclear how genetic predispositions promote neuroinflammation in MS and EAE. Here, we investigated how partial loss-of-function of suppressor of MEK1 (SMEK1), a regulatory subunit of protein phosphatase 4, facilitates the onset of MS and EAE.

Methods

C57BL/6 mice were immunized with myelin oligodendrocyte glycoprotein 35-55 (MOG_35-55) to establish the EAE model. Clinical signs were recorded and pathogenesis was investigated after immunization. CNS tissues were analyzed by immunostaining, quantitative polymerase chain reaction (qPCR), western blot analysis, and enzyme-linked immunosorbent assay (ELISA). Single-cell analysis was carried out in the cortices and hippocampus. Splenic and lymph node cells were evaluated with flow cytometry, qPCR, and western blot analysis.

Results

Here, we showed that partial Smek1 deficiency caused more severe symptoms in the EAE model than in controls by activating myeloid cells and that Smek1 was required for maintaining immunosuppressive function by modulating the indoleamine 2,3-dioxygenase (IDO1)-aryl hydrocarbon receptor (AhR) pathway. Single-cell sequencing and an in vitro study showed that Smek1-deficient microglia and macrophages were preactivated at steady state. After MOG_35-55 immunization, microglia and macrophages underwent hyperactivation and produced increased IL-1β in Smek1^-/+ mice at the peak stage. Moreover, dysfunction of the IDO1-AhR pathway resulted from the reduction of interferon γ (IFN-γ), enhanced antigen presentation ability, and inhibition of anti-inflammatory processes in Smek1^-/+ EAE mice.

Conclusions

The present study suggests a protective role of Smek1 in autoimmune demyelination pathogenesis via immune suppression and inflammation regulation in both the immune system and the central nervous system. Our findings provide an instructive basis for the roles of Smek1 in EAE and broaden the understanding of the genetic factors involved in the pathogenesis of autoimmune demyelination.

Available only for authorised users

Cohen PT, Philp A, Vazquez-Martin C. Protein phosphatase 4--from obscurity to vital functions. FEBS Lett. 2005;579(15):3278–86. https://doi.org/10.1016/j.febslet.2005.04.070.CrossRefPubMed

Liao FH, Hsiao WY, Lin YC, Chan YC, Huang CY. T cell proliferation and adaptive immune responses are critically regulated by protein phosphatase 4. Cell Cycle. 2016;15(8):1073–83. https://doi.org/10.1080/15384101.2016.1156267.CrossRefPubMedPubMedCentral

Liao FH, Shui JW, Hsing EW, Hsiao WY, Lin YC, Chan YC, et al. Protein phosphatase 4 is an essential positive regulator for Treg development, function, and protective gut immunity. Cell Biosci. 2014;4(1):25. https://doi.org/10.1186/2045-3701-4-25.CrossRefPubMedPubMedCentral

Mendoza MC, Booth EO, Shaulsky G, Firtel RA. MEK1 and protein phosphatase 4 coordinate Dictyostelium development and chemotaxis. Mol Cell Biol. 2007;27(10):3817–27. https://doi.org/10.1128/MCB.02194-06.CrossRefPubMedPubMedCentral

Chang WH, Choi SH, Moon BS, Cai M, Lyu J, Bai J, et al. Smek1/2 is a nuclear chaperone and cofactor for cleaved Wnt receptor Ryk, regulating cortical neurogenesis. Proc Natl Acad Sci U S A. 2017;114(50):E10717–25. https://doi.org/10.1073/pnas.1715772114.CrossRefPubMedPubMedCentral

Moon BS, Yun HM, Chang WH, Steele BH, Cai M, Choi SH, et al. Smek promotes corticogenesis through regulating Mbd3's stability and Mbd3/NuRD complex recruitment to genes associated with neurogenesis. PLoS Biol. 2017;15(5):e2001220. https://doi.org/10.1371/journal.pbio.2001220.CrossRefPubMedPubMedCentral

Lyu J, Kim HR, Yamamoto V, Choi SH, Wei Z, Joo CK, et al. Protein phosphatase 4 and Smek complex negatively regulate Par3 and promote neuronal differentiation of neural stem/progenitor cells. Cell Rep. 2013;5(3):593–600. https://doi.org/10.1016/j.celrep.2013.09.034.CrossRefPubMed

Sen I, Zhou X, Chernobrovkin A, Puerta-Cavanzo N, Kanno T, Salignon J, et al. DAF-16/FOXO requires Protein Phosphatase 4 to initiate transcription of stress resistance and longevity promoting genes. Nat Commun. 2020;11(1):138. https://doi.org/10.1038/s41467-019-13931-7.CrossRefPubMedPubMedCentral

Bogie JF, Stinissen P, Hendriks JJ. Macrophage subsets and microglia in multiple sclerosis. Acta Neuropathol. 2014;128(2):191–213. https://doi.org/10.1007/s00401-014-1310-2.CrossRefPubMed

10.

Lucchinetti C, Bruck W, Parisi J, Scheithauer B, Rodriguez M, Lassmann H. Heterogeneity of multiple sclerosis lesions: implications for the pathogenesis of demyelination. Ann Neurol. 2000;47(6):707–17. https://doi.org/10.1002/1531-8249(200006)47:6<707::AID-ANA3>3.0.CO;2-Q.CrossRefPubMed

11.

Singh S, Metz I, Amor S, van der Valk P, Stadelmann C, Bruck W. Microglial nodules in early multiple sclerosis white matter are associated with degenerating axons. Acta Neuropathol. 2013;125(4):595–608. https://doi.org/10.1007/s00401-013-1082-0.CrossRefPubMedPubMedCentral

12.

Zrzavy T, Hametner S, Wimmer I, Butovsky O, Weiner HL, Lassmann H. Loss of ‘homeostatic’ microglia and patterns of their activation in active multiple sclerosis. Brain. 2017;140(7):1900–13. https://doi.org/10.1093/brain/awx113.CrossRefPubMedPubMedCentral

13.

Gosselin D, Skola D, Coufal NG, Holtman IR, Schlachetzki JCM, Sajti E, et al. An environment-dependent transcriptional network specifies human microglia identity. Science. 2017;356(6344):eaal3222. https://doi.org/10.1126/science.aal3222.CrossRefPubMedPubMedCentral

14.

Sosa RA, Murphey C, Ji N, Cardona AE, Forsthuber TG. The kinetics of myelin antigen uptake by myeloid cells in the central nervous system during experimental autoimmune encephalomyelitis. J Immunol. 2013;191(12):5848–57. https://doi.org/10.4049/jimmunol.1300771.CrossRefPubMed

15.

Merson TD, Binder MD, Kilpatrick TJ. Role of cytokines as mediators and regulators of microglial activity in inflammatory demyelination of the CNS. NeuroMolecular Med. 2010;12(2):99–132. https://doi.org/10.1007/s12017-010-8112-z.CrossRefPubMed

16.

Billiau A, Heremans H, Vandekerckhove F, Dijkmans R, Sobis H, Meulepas E, et al. Enhancement of experimental allergic encephalomyelitis in mice by antibodies against IFN-gamma. J Immunol. 1988;140:1506–10.CrossRefPubMed

17.

Panitch HS, Robert LH, John S, Kenneth PJ. Treatment of multiple sclerosis with gamma interferon: exacerbations associated with activation of the immune system. Neurology. 1987;37:1097–102.CrossRefPubMed

18.

Skurkovich S, Boiko A, Beliaeva I, Buglak A, Alekseeva T, Smirnova N, et al. Randomized study of antibodies to IFN-gamma and TNF-alpha in secondary progressive multiple sclerosis. Mult Scler. 2001;7(5):277–84. https://doi.org/10.1177/135245850100700502.CrossRefPubMed

19.

Voorthuis JA, Uitdehaag BM, De Groot CJ, Goede PH, van der Meide PH, Dijkstra CD. Suppression of experimental allergic encephalomyelitis by intraventricular administration of interferon-gamma in Lewis rats. Clin Exp Immunol. 1990;81(2):183–8. https://doi.org/10.1111/j.1365-2249.1990.tb03315.x.CrossRefPubMedPubMedCentral

20.

Heremans H, Dillen C, Groenen M, Martens E, Billiau A. Chronic relapsing experimental autoimmune encephalomyelitis (CREAE) in mice: enhancement by monoclonal antibodies against interferon-gamma. Eur J Immunol. 1996;26(10):2393–8. https://doi.org/10.1002/eji.1830261019.CrossRefPubMed

21.

Lublin FD, Knobler RL, Kalman B, Goldhaber M, Marini J, Perrault M, et al. Monoclonal anti-gamma interferon antibodies enhance experimental allergic encephalomyelitis. Autoimmunity. 1993;16(4):267–74. https://doi.org/10.3109/08916939309014645.CrossRefPubMed

22.

Ferber IA, Brocke S, Taylor-Edwards C, Ridgway W, Dinisco C, Steinman L, et al. Mice with a disrupted IFN-gamma gene are susceptible to the induction of experimental autoimmune encephalomyelitis (EAE). J Immunol. 1996;156:5–7.CrossRefPubMed

23.

Bonney S, Seitz S, Ryan CA, Jones KL, Clarke P, Tyler KL, et al. Gamma interferon alters junctional integrity via Rho kinase, resulting in blood-brain barrier leakage in experimental viral encephalitis. mBio. 2019;10.

24.

Kelchtermans H, Billiau A, Matthys P. How interferon-gamma keeps autoimmune diseases in check. Trends Immunol. 2008;29(10):479–86. https://doi.org/10.1016/j.it.2008.07.002.CrossRefPubMed

25.

Popko B, Corbin JG, Baerwald KD, Dupree J, Garcia AM. The effects of interferon-gamma on the central nervous system. Mol Neurobiol. 1997;14(1-2):19–35. https://doi.org/10.1007/BF02740619.CrossRefPubMedPubMedCentral

26.

Smith BC, Sinyuk M, Jenkins JE, Psenicka MW, Williams JL. The impact of regional astrocyte interferon-γ signaling during chronic autoimmunity: a novel role for the immunoproteasome. J Neuroinflammation. 2020;17(1):184. https://doi.org/10.1186/s12974-020-01861-x.CrossRefPubMedPubMedCentral

27.

Xiao BG, Wu XC, Yang JS, Xu LY, Liu X, Huang YM, et al. Therapeutic potential of IFN-gamma-modified dendritic cells in acute and chronic experimental allergic encephalomyelitis. Int Immunol. 2004;16(1):13–22. https://doi.org/10.1093/intimm/dxh003.CrossRefPubMed

28.

Amani H, Shahbazi MA, D'Amico C, Fontana F, Abbaszadeh S, Santos HA. Microneedles for painless transdermal immunotherapeutic applications. J Control Release. 2021;330:185–217. https://doi.org/10.1016/j.jconrel.2020.12.019.CrossRefPubMed

29.

Wang S, Qi Y, Gao X, Qiu W, Liu Q, Guo X, et al. Hypoxia-induced lncRNA PDIA3P1 promotes mesenchymal transition via sponging of miR-124-3p in glioma. Cell Death Dis. 2020;11(3):168. https://doi.org/10.1038/s41419-020-2345-z.CrossRefPubMedPubMedCentral

30.

Duan R, Liu Q, Li J, Bian X, Yuan Q, Li Y, et al. A de novo frameshift mutation in TNFAIP3 impairs A20 deubiquitination function to cause neuropsychiatric systemic lupus erythematosus. J Clin Immunol. 2019;39(8):795–804. https://doi.org/10.1007/s10875-019-00695-4.CrossRefPubMed

31.

Hayashi S, Lewis P, Pevny L, McMahon AP. Efficient gene modulation in mouse epiblast using a Sox2Cre transgenic mouse strain. Mech Dev. 2002;119(Suppl 1):S97–S101. https://doi.org/10.1016/S0925-4773(03)00099-6.CrossRefPubMed

32.

Campolo F, Gori M, Favaro R, Nicolis S, Pellegrini M, Botti F, et al. Essential role of Sox2 for the establishment and maintenance of the germ cell line. Stem Cells. 2013;31(7):1408–21. https://doi.org/10.1002/stem.1392.CrossRefPubMed

33.

Easley-Neal C, Foreman O, Sharma N, Zarrin AA, Weimer RM. CSF1R ligands IL-34 and CSF1 are differentially required for microglia development and maintenance in white and gray matter brain regions. Front Immunol. 2019;10:2199. https://doi.org/10.3389/fimmu.2019.02199.CrossRefPubMedPubMedCentral

34.

van der Poel M, Ulas T, Mizee MR, Hsiao CC, Miedema SSM, Adelia, et al. Transcriptional profiling of human microglia reveals grey-white matter heterogeneity and multiple sclerosis-associated changes. Nat Commun. 2019;10(1):1139. https://doi.org/10.1038/s41467-019-08976-7.CrossRefPubMedPubMedCentral

35.

Chitu V, Stanley ER. Regulation of embryonic and postnatal development by the CSF-1 receptor. Curr Top Dev Biol. 2017;123:229–75. https://doi.org/10.1016/bs.ctdb.2016.10.004.CrossRefPubMed

36.

Wylot B, Mieczkowski J, Niedziolka S, Kaminska B, Zawadzka M. Csf1 deficiency dysregulates glial responses to demyelination and disturbs CNS white matter remyelination. Cells. 2019;9(1). https://doi.org/10.3390/cells9010099.

37.

Borjini N, Fernandez M, Giardino L, Calza L. Cytokine and chemokine alterations in tissue, CSF, and plasma in early presymptomatic phase of experimental allergic encephalomyelitis (EAE), in a rat model of multiple sclerosis. J Neuroinflammation. 2016;13(1):291. https://doi.org/10.1186/s12974-016-0757-6.CrossRefPubMedPubMedCentral

38.

Gushchina S, Pryce G, Yip PK, Wu D, Pallier P, Giovannoni G, et al. Increased expression of colony-stimulating factor-1 in mouse spinal cord with experimental autoimmune encephalomyelitis correlates with microglial activation and neuronal loss. Glia. 2018;66(10):2108–25. https://doi.org/10.1002/glia.23464.CrossRefPubMed

39.

Yan X, Maixner DW, Li F, Weng HR. Chronic pain and impaired glial glutamate transporter function in lupus-prone mice are ameliorated by blocking macrophage colony-stimulating factor-1 receptors. J Neurochem. 2017;140(6):963–76. https://doi.org/10.1111/jnc.13952.CrossRefPubMedPubMedCentral

40.

Hwu P, Du MX, Lapointe R, Do M, Taylor MW, Young HA. Indoleamine 2,3-dioxygenase production by human dendritic cells results in the inhibition of T cell proliferation. J Immunol. 2000;164(7):3596–9. https://doi.org/10.4049/jimmunol.164.7.3596.CrossRefPubMed

41.

Platten M, Nollen EAA, Rohrig UF, Fallarino F, Opitz CA. Tryptophan metabolism as a common therapeutic target in cancer, neurodegeneration and beyond. Nat Rev Drug Discov. 2019;18(5):379–401. https://doi.org/10.1038/s41573-019-0016-5.CrossRefPubMed

42.

Opitz CA, Litzenburger UM, Sahm F, Ott M, Tritschler I, Trump S, et al. An endogenous tumour-promoting ligand of the human aryl hydrocarbon receptor. Nature. 2011;478(7368):197–203. https://doi.org/10.1038/nature10491.CrossRefPubMed

43.

Apetoh L, Quintana FJ, Pot C, Joller N, Xiao S, Kumar D, et al. The aryl hydrocarbon receptor interacts with c-Maf to promote the differentiation of type 1 regulatory T cells induced by IL-27. Nat Immunol. 2010;11(9):854–61. https://doi.org/10.1038/ni.1912.CrossRefPubMedPubMedCentral

44.

Piper CJM, Rosser EC, Oleinika K, Nistala K, Krausgruber T, Rendeiro AF, et al. Aryl hydrocarbon receptor contributes to the transcriptional program of IL-10-producing regulatory B cells. Cell Rep. 2019;29(7):1878–92 e1877. https://doi.org/10.1016/j.celrep.2019.10.018.CrossRefPubMedPubMedCentral

45.

Adams O, Besken K, Oberdorfer C, MacKenzie CR, Takikawa O, Daubener W. Role of indoleamine-2,3-dioxygenase in alpha/beta and gamma interferon-mediated antiviral effects against herpes simplex virus infections. J Virol. 2004;78(5):2632–6. https://doi.org/10.1128/JVI.78.5.2632-2636.2004.CrossRefPubMedPubMedCentral

46.

Du HX, Chen XG, Zhang L, Liu Y, Zhan CS, Chen J, et al. Microglial activation and neurobiological alterations in experimental autoimmune prostatitis-induced depressive-like behavior in mice. Neuropsychiatr Dis Treat. 2019;15:2231–45. https://doi.org/10.2147/NDT.S211288.CrossRefPubMedPubMedCentral

47.

Garrison AM, Parrott JM, Tunon A, Delgado J, Redus L, O'Connor JC. Kynurenine pathway metabolic balance influences microglia activity: Targeting kynurenine monooxygenase to dampen neuroinflammation. Psychoneuroendocrinology. 2018;94:1–10. https://doi.org/10.1016/j.psyneuen.2018.04.019.CrossRefPubMedPubMedCentral

48.

Arellano G, Ottum PA, Reyes LI, Burgos PI, Naves R. Stage-specific role of interferon-gamma in experimental autoimmune encephalomyelitis and multiple sclerosis. Front Immunol. 2015;6. https://doi.org/10.3389/fimmu.2015.00492.

49.

Rozman P, Svajger U. The tolerogenic role of IFN-gamma. Cytokine Growth Factor Rev. 2018;41:40–53. https://doi.org/10.1016/j.cytogfr.2018.04.001.CrossRefPubMed

Title: Smek1 deficiency exacerbates experimental autoimmune encephalomyelitis by activating proinflammatory microglia and suppressing the IDO1-AhR pathway
Authors: Ruo-Nan Duan
Chun-Lin Yang
Tong Du
Ai Liu
An-Ran Wang
Wen-Jie Sun
Xi Li
Jiang-Xia Li
Chuan-Zhu Yan
Qi-Ji Liu
Publication date: 01-12-2021
Publisher: BioMed Central
Keyword: Multiple Sclerosis
Published in: Journal of Neuroinflammation / Issue 1/2021
Electronic ISSN: 1742-2094
DOI: https://doi.org/10.1186/s12974-021-02193-0

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Smek1 deficiency exacerbates experimental autoimmune encephalomyelitis by activating proinflammatory microglia and suppressing the IDO1-AhR pathway

Abstract

Background

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2021

LncRNA HOXA-AS2 regulates microglial polarization via recruitment of PRC2 and epigenetic modification of PGC-1α expression

Systemic inflammation suppresses spinal respiratory motor plasticity via mechanisms that require serine/threonine protein phosphatase activity

Downregulation of metallothionein-2 contributes to oxaliplatin-induced neuropathic pain

Inflammation after spinal cord injury: a review of the critical timeline of signaling cues and cellular infiltration

CXCR5 down-regulation alleviates cognitive dysfunction in a mouse model of sepsis-associated encephalopathy: potential role of microglial autophagy and the p38MAPK/NF-κB/STAT3 signaling pathway

miR-1224 contributes to ischemic stroke-mediated natural killer cell dysfunction by targeting Sp1 signaling