Abstract
Age at onset in spinocerebellar ataxia type 3 (SCA3/MJD) is incompletely explained by the size of the CAG tract at the ATXN3 gene, implying the existence of genetic modifiers. A role of inflammation in SCA3 has been postulated, involving altered cytokines levels; promoter variants leading to alterations in cytokines expression could influence onset. Using blood from 86 SCA3 patients and 106 controls, this work aimed to analyse promoter variation of four cytokines (IL1A, IL1B, IL6 and TNF) and to investigate the association between variants detected and their transcript levels, evaluated by quantitative PCR. Moreover, the effect of APOE isoforms, known to modulate cytokines, was investigated. Correlations between cytokine variants and onset were tested; the cumulative modifier effects of cytokines and APOE were analysed. Patients carrying the IL6*C allele had a significant earlier onset (4 years in average) than patients carrying the G allele, in agreement with lower mRNA levels produced by IL6*C carriers. The presence of APOE*ɛ2 allele seems to anticipate onset in average 10 years in patients carrying the IL6*C allele; a larger number of patients will be needed to confirm this result. These results highlight the pertinence of conducting further research on the role of cytokines as SCA3 modulators, pointing to the presence of shared mechanisms involving IL6 and APOE.
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References
Bettencourt, C., Fialho, R. N., Santos, C., Montiel, R., Bruges-Armas, J., MacIel, P., & Lima, M. (2008). Segregation distortion of wild-type alleles at the Machado–Joseph disease locus: A study in normal families from the Azores islands (Portugal). Journal of Human Genetics, 53(4), 333–339. doi:10.1007/s10038-008-0261-7.
Bettencourt, C., & Lima, M. (2011). Machado–Joseph Disease: From first descriptions to new perspectives. Orphanet Journal of Rare Diseases, 6, 35. doi:10.1186/1750-1172-6-35.
Bettencourt, C., Raposo, M., Kazachkova, N., Cymbron, T., Santos, C., Kay, T., et al. (2011). The APOE ε2 allele increases the risk of earlier age at onset in Machado–Joseph disease. Archives of Neurology, 68(12), 1580–1583. doi:10.1001/archneurol.2011.636.
da Silva Carvalho, G., Saute, J., Haas, C., Torrez, V., Brochier, A., Souza, G., et al. (2015). Cytokines in Machado–Joseph disease/spinocerebellar ataxia 3. Cerebellum (London, England). doi:10.1007/s12311-015-0719-z.
Dominici, R., Cattaneo, M., Malferrari, G., Archi, D., Mariani, C., Grimaldi, L., & Biunno, I. (2002). Cloning and functional analysis of the allelic polymorphism in the transcription regulatory region of interleukin-1 alpha. Immunogenetics, 54(2), 82–86. doi:10.1007/s00251-002-0445-9.
Evers, M. M., Toonen, L. J. A., & van Roon-Mom, W. M. C. (2014). Ataxin-3 protein and RNA toxicity in spinocerebellar ataxia type 3: Current insights and emerging therapeutic strategies. Molecular Neurobiology, 49(3), 1513–1531. doi:10.1007/s12035-013-8596-2.
Evert, B., Schelhaas, J., Fleischer, H., de Vos, R., Brunt, E., Stenzel, W., et al. (2006). Neuronal intranuclear inclusions, dysregulation of cytokine expression and cell death in spinocerebellar ataxia type 3. Clinical Neuropathology, 25(6), 272–281.
Evert, B., Vogt, I., Kindermann, C., Ozimek, L., de Vos, R., Brunt, E., et al. (2001). Inflammatory genes are upregulated in expanded ataxin-3-expressing cell lines and spinocerebellar ataxia type 3 brains. Journal of Neuroscience, 21(15), 5389–5396.
Evert, B., Vogt, I., Vieira-Saecker, A., Ozimek, L., de Vos, R., Brunt, E., et al. (2003). Gene expression profiling in ataxin-3 expressing cell lines reveals distinct effects of normal and mutant ataxin-3. Journal of Neuropathology and Experimental Neurology, 62(10), 1006–1018.
Fishman, D., Faulds, G., Jeffery, R., Mohamed-Ali, V., Yudkin, J., Humphries, S., & Woo, P. (1998). The effect of novel polymorphisms in the interleukin-6 (IL-6) gene on IL-6 transcription and plasma IL-6 levels, and an association with systemic-onset juvenile chronic arthritis. Journal of Clinical Investigation, 102(7), 1369–1376. doi:10.1172/JCI2629.
França, M. C., Emmel, V. E., D’Abreu, A., Maurer-Morelli, C. V., Secolin, R., Bonadia, L. C., et al. (2012). Normal ATXN3 allele but not CHIP polymorphisms modulates age at onset in Machado–Joseph disease. Frontiers in Neurology, 3, 164. doi:10.3389/fneur.2012.00164.
Hall, S., Perregaux, D., Gabel, C., Woodworth, T., Durham, L., Huizinga, T., et al. (2004). Correlation of polymorphic variation in the promoter region of the interleukin-1 beta gene with secretion of interleukin-1 beta protein. Arthritis and Rheumatism, 50(6), 1976–1983. doi:10.1002/art.20310.
IBM Corp. (2013). IBM SPSS statistics for windows, Version 220. Armonk, NY: IBM Corp.
Jardim, L., Silveira, I., Pereira, M. L., do Céu Moreira, M., Mendonça, P., Sequeiros, J., & Giugliani, R. (2003). Searching for modulating effects of SCA2, SCA6 and DRPLA CAG tracts on the Machado-Joseph disease (SCA3) phenotype. Acta Neurologica Scandinavica, 107(3), 211–214.
Long, Z., Chen, Z., Wang, C., Huang, F., Peng, H., Hou, X., et al. (2015). Two novel SNPs in ATXN3 3′ UTR may decrease age at onset of SCA3/MJD in Chinese patients. PLoS ONE, 10(2), e0117488. doi:10.1371/journal.pone.0117488.
Maciel, P., Costa, M. C., Ferro, A., Rousseau, M., Santos, C. S., Gaspar, C., et al. (2001). Improvement in the molecular diagnosis of Machado–Joseph disease. Archives of Neurology, 58(11), 1821–1827. doi:10.1001/archneur.58.11.1821.
Nishimura, M., Kawakami, H., Maruyama, H., Izumi, Y., Kuno, S., Kaji, R., & Nakamura, S. (2001). Influence of interleukin-1beta gene polymorphism on age-at-onset of spinocerebellar ataxia 6 (SCA6) in Japanese patients. Neuroscience Letters, 307(2), 128–130.
Olejniczak, M., Urbanek, M., & Krzyzosiak, W. (2015). The role of the immune system in triplet repeat expansion diseases. Mediators of Inflammation, 2015, 1–11. doi:10.1155/2015/873860.
Peng, H., Wang, C., Chen, Z., Sun, Z., Jiao, B., Li, K., et al. (2014). APOE ε2 allele may decrease the age at onset in patients with spinocerebellar ataxia type 3 or Machado–Joseph disease from the Chinese Han population. Neurobiology of Aging, 35(9), 2179-e15. doi:10.1016/j.neurobiolaging.2014.03.020.
Raposo, M., Bettencourt, C., Maciel, P., Gao, F., Ramos, A., Kazachkova, N., et al. (2015a). Novel candidate blood-based transcriptional biomarkers of Machado–Joseph disease. Movement Disorders, 30(7), 968–975. doi:10.1002/mds.26238.
Raposo, M., Ramos, A., Bettencourt, C., & Lima, M. (2015b). Replicating studies of genetic modifiers in spinocerebellar ataxia type 3: Can homogeneous cohorts aid? Brain. doi:10.1093/brain/awv206.
Siebert, M., Donis, K. C., Socal, M., Rieder, C., Emmel, V. E., Vairo, F., et al. (2012). Glucocerebrosidase gene variants in parkinsonian patients with Machado–Joseph/spinocerebellar ataxia 3. Parkinsonism and Related Disorders, 18(2), 185–190. doi:10.1016/j.parkreldis.2011.09.024.
Tezenas du Montcel, S., Durr, A., Bauer, P., Figueroa, K., Ichikawa, Y., Brussino, A., et al. (2014). Modulation of the age at onset in spinocerebellar ataxia by CAG tracts in various genes. Brain, 137(Pt 9), 2444–2455. doi:10.1093/brain/awu174.
Wilson, A. G., Symons, J. A., McDowell, T. L., McDevitt, H. O., & Duff, G. W. (1997). Effects of a polymorphism in the human tumor necrosis factor alpha promoter on transcriptional activation. Proceedings of the National Academy of Sciences of the United States of America, 94(7), 3195–3199.
Zhang, H., Wu, L.-M., & Wu, J. (2011). Cross-talk between apolipoprotein E and cytokines. Mediators of Inflammation, 2011, 1–10. doi:10.1155/2011/949072.
Acknowledgments
This work was funded by FEDER funds through the Operational Competitiveness Programme—COMPETE and by National Funds through FCT—Fundação para a Ciência e a Tecnologia under the project FCOMP-01-0124-FEDER-028753 (PTDC/DTP/PIC/0370/2012). A PhD fellowship M3.1.2/F/006/2011 (MR) and postdoctoral fellowships M3.1.7/F/031/2011 (AR) and M3.1.7/F/002/2008 (NK) were supported by Fundo Regional para a Ciência (FRC), Governo dos Açores. CB was supported by the Wellcome Trust (UK).
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12017_2016_8416_MOESM1_ESM.docx
Supplementary Table 1. Multiplex PCR primers, size of amplified fragments, restriction enzymes, size of restriction fragments as well as PCR-RFLP reactions mixture and conditions (DOCX 21 kb)
12017_2016_8416_MOESM2_ESM.docx
Supplementary Table 2. Genotypic, allelic frequencies and p values for the Hardy–Weinberg equilibrium test for each cytokine loci studied in all and unrelated SCA3 patients and population-matched controls. Differentiation exact test p values are also shown (DOCX 19 kb)
12017_2016_8416_MOESM3_ESM.docx
Supplementary Table 3. Genetic and clinical features of the 86 SCA3 patients divided by cytokines alleles, as well as presence/absence of APOE*ɛ2 allele (DOCX 17 kb)
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Raposo, M., Bettencourt, C., Ramos, A. et al. Promoter Variation and Expression Levels of Inflammatory Genes IL1A, IL1B, IL6 and TNF in Blood of Spinocerebellar Ataxia Type 3 (SCA3) Patients. Neuromol Med 19, 41–45 (2017). https://doi.org/10.1007/s12017-016-8416-8
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DOI: https://doi.org/10.1007/s12017-016-8416-8