Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Glycogen synthase kinase 3 controls T-cell exhaustion by regulating NFAT activation

Cellular & Molecular Immunology volume 20, pages 1127–1139 (2023)Cite this article

Subjects

Abstract

Cellular immunity mediated by CD8+ T cells plays an indispensable role in bacterial and viral clearance and cancers. However, persistent antigen stimulation of CD8+ T cells leads to an exhausted or dysfunctional cellular state characterized by the loss of effector function and high expression of inhibitory receptors during chronic viral infection and in tumors. Numerous studies have shown that glycogen synthase kinase 3 (GSK3) controls the function and development of immune cells, but whether GSK3 affects CD8+ T cells is not clearly elucidated. Here, we demonstrate that mice with deletion of Gsk3α and Gsk3β in activated CD8+ T cells (DKO) exhibited decreased CTL differentiation and effector function during acute and chronic viral infection. In addition, DKO mice failed to control tumor growth due to the upregulated expression of inhibitory receptors and augmented T-cell exhaustion in tumor-infiltrating CD8+ T cells. Strikingly, anti-PD-1 immunotherapy substantially restored tumor rejection in DKO mice. Mechanistically, GSK3 regulates T-cell exhaustion by suppressing TCR-induced nuclear import of NFAT, thereby in turn dampening NFAT-mediated exhaustion-related gene expression, including TOX/TOX2 and PD-1. Thus, we uncovered the molecular mechanisms underlying GSK3 regulation of CTL differentiation and T-cell exhaustion in anti-tumor immune responses.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

Data availability

The data that support the findings of this study are available from the corresponding authors upon request.

References

  1. Chen DS, Mellman I. Oncology meets immunology: the cancer-immunity cycle. Immunity. 2013;39:1–10.

    Article  PubMed  Google Scholar 

  2. Denton AE, et al. Differentiation-dependent functional and epigenetic landscapes for cytokine genes in virus-specific CD8 + T cells. Proc Natl Acad Sci USA. 2011;108:15306–11.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Jenkins MR, et al. Cell cycle-related acquisition of cytotoxic mediators defines the progressive differentiation to effector status for virus-specific CD8 + T cells. J Immunol. 2008;181:3818–22.

    Article  CAS  PubMed  Google Scholar 

  4. Wherry EJ. T cell exhaustion. Nat Immunol. 2011;12:492–9.

    Article  CAS  PubMed  Google Scholar 

  5. Doering TA, et al. Network analysis reveals centrally connected genes and pathways involved in CD8 + T cell exhaustion versus memory. Immunity. 2012;37:1130–44.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Day CL, et al. PD-1 expression on HIV-specific T cells is associated with T-cell exhaustion and disease progression. Nature. 2006;443:350–4.

    Article  CAS  PubMed  Google Scholar 

  7. Wherry EJ, Kurachi M. Molecular and cellular insights into T cell exhaustion. Nat Rev Immunol. 2015;15:486–99.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Topalian SL, Drake CG, Pardoll DM. Immune checkpoint blockade: a common denominator approach to cancer therapy. Cancer Cell. 2015;27:450–61.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. He R, et al. PD-1 and CTLA-4 inhibitors in combination vs. alone for the treatment of advanced melanoma: a systematic review and meta-analysis. Med (Baltim). 2022;101:e30561.

    Article  CAS  Google Scholar 

  10. Nguyen LT, Ohashi PS. Clinical blockade of PD1 and LAG3–potential mechanisms of action. Nat Rev Immunol. 2015;15:45–56.

    Article  CAS  PubMed  Google Scholar 

  11. Araki K, Youngblood B, Ahmed R. Programmed cell death 1-directed immunotherapy for enhancing T-cell function. Cold Spring Harb Symp Quant Biol. 2013;78:239–47.

    Article  PubMed  Google Scholar 

  12. Martinez GJ, et al. The transcription factor NFAT promotes exhaustion of activated CD8( + ) T cells. Immunity. 2015;42:265–78.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Man K, et al. Transcription factor IRF4 promotes CD8( + ) T cell exhaustion and limits the development of memory-like T cells during chronic infection. Immunity. 2017;47:1129–41 e5.

    Article  CAS  PubMed  Google Scholar 

  14. Chen Y, et al. BATF regulates progenitor to cytolytic effector CD8( + ) T cell transition during chronic viral infection. Nat Immunol. 2021;22:996–1007.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Seo H, et al. BATF and IRF4 cooperate to counter exhaustion in tumor-infiltrating CAR T cells. Nat Immunol. 2021;22:983–95.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Tsui C, et al. MYB orchestrates T cell exhaustion and response to checkpoint inhibition. Nature. 2022;609:354–60.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Muller MR, Rao A. NFAT, immunity and cancer: a transcription factor comes of age. Nat Rev Immunol. 2010;10:645–56.

    Article  PubMed  Google Scholar 

  18. Seo H, et al. TOX and TOX2 transcription factors cooperate with NR4A transcription factors to impose CD8( + ) T cell exhaustion. Proc Natl Acad Sci USA. 2019;116:12410–12415.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Khan O, et al. TOX transcriptionally and epigenetically programs CD8( + ) T cell exhaustion. Nature. 2019;571:211–18.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Scott AC, et al. TOX is a critical regulator of tumour-specific T cell differentiation. Nature. 2019;571:270–74.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Alfei F, et al. TOX reinforces the phenotype and longevity of exhausted T cells in chronic viral infection. Nature. 2019;571:265–69.

    Article  CAS  PubMed  Google Scholar 

  22. Yao C, et al. Single-cell RNA-seq reveals TOX as a key regulator of CD8( + ) T cell persistence in chronic infection. Nat Immunol. 2019;20:890–901.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Fiol CJ, et al. Formation of protein kinase recognition sites by covalent modification of the substrate. Molecular mechanism for the synergistic action of casein kinase II and glycogen synthase kinase 3. J Biol Chem. 1987;262:14042–8.

    Article  CAS  PubMed  Google Scholar 

  24. Frame S, Cohen P, Biondi RM. A common phosphate binding site explains the unique substrate specificity of GSK3 and its inactivation by phosphorylation. Mol Cell. 2001;7:1321–7.

    Article  CAS  PubMed  Google Scholar 

  25. ter Haar E, et al. Structure of GSK3beta reveals a primed phosphorylation mechanism. Nat Struct Biol. 2001;8:593–6.

    Article  PubMed  Google Scholar 

  26. Ying QL, et al. The ground state of embryonic stem cell self-renewal. Nature. 2008;453:519–23.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Beals CR, et al. Nuclear export of NF-ATc enhanced by glycogen synthase kinase-3. Science. 1997;275:1930–4.

    Article  CAS  PubMed  Google Scholar 

  28. Beurel E, Michalek SM, Jope RS. Innate and adaptive immune responses regulated by glycogen synthase kinase-3 (GSK3). Trends Immunol. 2010;31:24–31.

    Article  CAS  PubMed  Google Scholar 

  29. Ohteki T, et al. Negative regulation of T cell proliferation and interleukin 2 production by the serine threonine kinase GSK-3. J Exp Med. 2000;192:99–104.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Taylor A, et al. Glycogen synthase kinase 3 inactivation drives T-bet-mediated downregulation of co-receptor PD-1 to enhance CD8(+) cytolytic T cell responses. Immunity. 2016;44:274–86.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Rudd CE, Chanthong K, Taylor A. Small molecule inhibition of GSK-3 specifically inhibits the transcription of inhibitory co-receptor LAG-3 for enhanced anti-tumor immunity. Cell Rep. 2020;30:2075–82 e4.

    Article  CAS  PubMed  Google Scholar 

  32. Pearce EL, et al. Control of effector CD8 + T cell function by the transcription factor Eomesodermin. Science. 2003;302:1041–3.

    Article  CAS  PubMed  Google Scholar 

  33. Intlekofer AM, et al. Effector and memory CD8 + T cell fate coupled by T-bet and eomesodermin. Nat Immunol. 2005;6:1236–44.

    Article  CAS  PubMed  Google Scholar 

  34. Liang C, et al. TOX as a potential target for immunotherapy in lymphocytic malignancies. Biomark Res. 2021;9:20.

    Article  PubMed  PubMed Central  Google Scholar 

  35. Clevers H, Nusse R. Wnt/beta-catenin signaling and disease. Cell. 2012;149:1192–205.

    Article  CAS  PubMed  Google Scholar 

  36. Liu C, et al. Glycogen synthase kinase 3 drives thymocyte egress by suppressing beta-catenin activation of Akt. Sci Adv. 2021;7:eabg6262.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Crabtree GR, Olson EN. NFAT signaling: choreographing the social lives of cells. Cell. 2002;109:S67–79.

    Article  CAS  PubMed  Google Scholar 

  38. Kurachi M, et al. The transcription factor BATF operates as an essential differentiation checkpoint in early effector CD8 + T cells. Nat Immunol. 2014;15:373–83.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Chen L, et al. Structure of the DNA-binding domains from NFAT, Fos and Jun bound specifically to DNA. Nature. 1998;392:42–8.

    Article  CAS  PubMed  Google Scholar 

  40. Duran J, et al. GSK-3beta/NFAT signaling is involved in testosterone-induced cardiac myocyte hypertrophy. PLoS One. 2016;11:e0168255.

    Article  PubMed  PubMed Central  Google Scholar 

  41. Kim MS, et al. Nerve growth factor (NGF) regulates activity of nuclear factor of activated T-cells (NFAT) in neurons via the phosphatidylinositol 3-kinase (PI3K)-Akt-glycogen synthase kinase 3beta (GSK3beta) pathway. J Biol Chem. 2014;289:31349–60.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank all members of the Liu Labs (Xiamen University) for the discussion and technical assistance. We thank Xiaohong Ma and Xiufeng Sun at the Xiamen University Flow Cytometry Core and Suqin Wu in the XMU Laboratory Animal Center for technical assistance.

Funding

This study was supported by the National Natural Science Foundation of China (31770953, 81830047, and 81961138008 to CX and 32070877 to W-HL), 1000 Young Talents Program of China (NX), and the Fundamental Research Funds for the Central Universities of China-Xiamen University (20720170064 to CX). The funders had no role in the study design, data collection, and analysis, decision to publish, or preparation of the manuscript.

Author information

Author notes

  1. Changchun Xiao

    Present address: Sanofi Institute for Biomedical Research, Suzhou, Jiangsu, 215123, China

Authors and Affiliations

  1. State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Faculty of Medicine and Life Science, Xiamen University, Xiamen, 361102, Fujian, China

    Yubing Fu, Jinjia Wang, Kunyu Liao, Xianjun Gao, Ronghan Tang, Binbin Fan, Yazhen Hong, Nengming Xiao & Wen-Hsien Liu

  2. Department of Cell Biology, School of Life Science, Anhui Medical University, Hefei, 230031, Anhui, China

    Chenfeng Liu

Authors
  1. Yubing Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jinjia Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Chenfeng Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kunyu Liao
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Xianjun Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ronghan Tang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Binbin Fan
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yazhen Hong
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Nengming Xiao
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Changchun Xiao
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Wen-Hsien Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

YF conceived the study. YF, JW, and CL performed most of the mouse, cell, and molecular biology experiments. YF and JW performed RNA sequencing experiments. RT and KL analyzed the RNA sequencing data. YF, JW, and XG performed several of the molecular experiments. W-HL cosupervised the project. YF and W-HL wrote the manuscript with input from all authors.

Corresponding authors

Correspondence to Yubing Fu or Wen-Hsien Liu.

Ethics declarations

Competing interests

The authors declare no competing interests.

Supplementary information

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Fu, Y., Wang, J., Liu, C. et al. Glycogen synthase kinase 3 controls T-cell exhaustion by regulating NFAT activation. Cell Mol Immunol 20, 1127–1139 (2023). https://doi.org/10.1038/s41423-023-01075-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41423-023-01075-0

Keywords

Search

Advanced search

Quick links