Abstract
CFTR, a chloride channel and ion channel regulator studied mostly in epithelial cells, has been reported to participate in immune regulation and likely affect the risk of cancer development. However, little is known about the effects of CFTR on the differentiation and function of γδ T cells. In this study, we observed that CFTR was functionally expressed on the cell surface of γδ T cells. Genetic deletion and pharmacological inhibition of CFTR both increased IFN-γ release by peripheral γδ T cells and potentiated the cytolytic activity of these cells against tumor cells both in vitro and in vivo. Interestingly, the molecular mechanisms underlying the regulation of γδ T cell IFN-γ production by CFTR were either TCR dependent or related to Ca2+ influx. CFTR was recruited to TCR immunological synapses and attenuated Lck-P38 MAPK-c-Jun signaling. In addition, CFTR was found to modulate TCR-induced Ca2+ influx and membrane potential (Vm)-induced Ca2+ influx and subsequently regulate the calcineurin-NFATc1 signaling pathway in γδ T cells. Thus, CFTR serves as a negative regulator of IFN-γ production in γδ T cells and the function of these cells in antitumor immunity. Our investigation suggests that modification of the CFTR activity of γδ T cells may be a potential immunotherapeutic strategy for cancer.
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References
Cahalan, M. D. & Chandy, K. G. The functional network of ion channels in T lymphocytes. Immunol. Rev. 231, 59–87 (2009).
Cai, X., Wang, X., Patel, S. & Clapham, D. E. Insights into the early evolution of animal calcium signaling machinery: a unicellular point of view. Cell Calcium 57, 166–173 (2015).
Clapham, D. E. Calcium signaling. Cell 131, 1047–1058 (2007).
Feske, S., Concepcion, A. R. & Coetzee, W. A. Eye on ion channels in immune cells. Sci. Signal. 12, 572 (2019).
Feske, S. Calcium signalling in lymphocyte activation and disease. Nat. Rev. Immunol. 7, 690–702 (2007).
Feske, S., Skolnik, E. Y. & Prakriya, M. Ion channels and transporters in lymphocyte function and immunity. Nat. Rev. Immunol. 12, 532–547 (2012).
Shaw, P. J., Qu, B., Hoth, M. & Feske, S. Molecular regulation of CRAC channels and their role in lymphocyte function. Cell. Mol. Life Sci. 70, 2637–2656 (2013).
Maul-Pavicic, A. et al. ORAI1-mediated calcium influx is required for human cytotoxic lymphocyte degranulation and target cell lysis. Proc. Natl Acad. Sci. USA 108, 3324–3329 (2011).
Shaw, P. J. & Feske, S. Regulation of lymphocyte function by ORAI and STIM proteins in infection and autoimmunity. J. Physiol. 590, 4157–4167 (2012).
Li, F. Y. et al. Second messenger role for Mg2+ revealed by human T-cell immunodeficiency. Nature 475, 471–476 (2011).
Chandy, K. G., DeCoursey, T. E., Cahalan, M. D., McLaughlin, C. & Gupta, S. Voltage-gated potassium channels are required for human T lymphocyte activation. J. Exp. Med. 160, 369–385 (1984).
DeCoursey, T. E., Chandy, K. G., Gupta, S. & Cahalan, M. D. Voltage-dependent ion channels in T-lymphocytes. J. Neuroimmunol. 10, 71–95 (1985).
Launay, P. et al. TRPM4 regulates calcium oscillations after T cell activation. Science 306, 1374–1377 (2004).
Launay, P. et al. TRPM4 is a Ca2+-activated nonselective cation channel mediating cell membrane depolarization. Cell 109, 397–407 (2002).
Chimote, A. A. et al. A defect in KCa3.1 channel activity limits the ability of CD8(+) T cells from cancer patients to infiltrate an adenosine-rich microenvironment. Sci. Signal. 11, 527 (2018).
Crottes, D. et al. Immature human dendritic cells enhance their migration through KCa3.1 channel activation. Cell Calcium 59, 198–207 (2016).
Kuras, Z., Yun, Y. H., Chimote, A. A., Neumeier, L. & Conforti, L. KCa3.1 and TRPM7 channels at the uropod regulate migration of activated human T cells. PloS ONE 7, e43859 (2012).
Riordan, J. R. et al. Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA. Science 245, 1066–1073 (1989).
Chen, J. H., Schulman, H. & Gardner, P. A cAMP-regulated chloride channel in lymphocytes that is affected in cystic fibrosis. Science 243, 657–660 (1989).
Shah, V. S. et al. Airway acidification initiates host defense abnormalities in cystic fibrosis mice. Science 351, 503–507 (2016).
Pankow, S. et al. F508 CFTR interactome remodelling promotes rescue of cystic fibrosis. Nature 528, 510–516 (2015).
Duan, Y. et al. Keratin K18 increases cystic fibrosis transmembrane conductance regulator (CFTR) surface expression by binding to its C-terminal hydrophobic patch. J. Biol. Chem. 287, 40547–40559 (2012).
Feske, S., Wulff, H. & Skolnik, E. Y. Ion channels in innate and adaptive immunity. Annu. Rev. Immunol. 33, 291–353 (2015).
Puga Molina, L. C. et al. CFTR/ENaC-dependent regulation of membrane potential during human sperm capacitation is initiated by bicarbonate uptake through NBC. J. Biol. Chem. 293, 9924–9936 (2018).
Wei, L. et al. The C-terminal part of the R-domain, but not the PDZ binding motif, of CFTR is involved in interaction with Ca(2+)-activated Cl- channels. Pflug. Arch. 442, 280–285 (2001).
Ogura, T. et al. ClC-3B, a novel ClC-3 splicing variant that interacts with EBP50 and facilitates expression of CFTR-regulated ORCC. FASEB J. 16, 863–865 (2002).
Welling, P. A. & Ho, K. A comprehensive guide to the ROMK potassium channel: form and function in health and disease. Am. J. Physiol. Ren. Physiol. 297, F849–F863 (2009).
Mueller, C. et al. Lack of cystic fibrosis transmembrane conductance regulator in CD3+ lymphocytes leads to aberrant cytokine secretion and hyperinflammatory adaptive immune responses. Am. J. Respir. Cell Mol. Biol. 44, 922–929 (2011).
Allard, J. B. et al. Aspergillus fumigatus generates an enhanced Th2-biased immune response in mice with defective cystic fibrosis transmembrane conductance regulator. J. Immunol. 177, 5186–5194 (2006).
Dorsey, J. & Gonska, T. Bacterial overgrowth, dysbiosis, inflammation, and dysmotility in the Cystic Fibrosis intestine. J. Cyst. Fibros. 16(Suppl 2), S14–S23 (2017).
Riquelme, S. A. et al. Cystic fibrosis transmembrane conductance regulator attaches tumor suppressor PTEN to the membrane and promotes anti pseudomonas aeruginosa immunity. Immunity 47, 1169–81 e7 (2017).
Holderness, J., Hedges, J. F., Ramstead, A. & Jutila, M. A. Comparative biology of gammadelta T cell function in humans, mice, and domestic animals. Annu. Rev. Anim. Biosci. 1, 99–124 (2013).
Chien, Y. H., Meyer, C. & Bonneville, M. gammadelta T cells: first line of defense and beyond. Annu. Rev. Immunol. 32, 121–155 (2014).
Serre, K. & Silva-Santos, B. Molecular mechanisms of differentiation of murine pro-inflammatory gammadelta T cell subsets. Front. Immunol. 4, 431 (2013).
Sutton, C. E. et al. Interleukin-1 and IL-23 induce innate IL-17 production from gammadelta T cells, amplifying Th17 responses and autoimmunity. Immunity 31, 331–341 (2009).
Ferrick, D. A. et al. Differential production of interferon-gamma and interleukin-4 in response to Th1- and Th2-stimulating pathogens by gamma delta T cells in vivo. Nature 373, 255–257 (1995).
Born, W. K., Yin, Z., Hahn, Y. S., Sun, D. & O’Brien, R. L. Analysis of gamma delta T cell functions in the mouse. J. Immunol. 184, 4055–4061 (2010).
Gao, Y. et al. Gamma delta T cells provide an early source of interferon gamma in tumor immunity. J. Exp. Med. 198, 433–442 (2003).
He, W. et al. Naturally activated V gamma 4 gamma delta T cells play a protective role in tumor immunity through expression of eomesodermin. J. Immunol. 185, 126–133 (2010).
Chen, L. et al. Epigenetic and transcriptional programs lead to default IFN-gamma production by gammadelta T cells. J. Immunol. 178, 2730–2736 (2007).
Lo Presti, E. et al. Current advances in gammadelta T cell-based tumor immunotherapy. Front. Immunol. 8, 1401 (2017).
Alnaggar, M. et al. Allogenic Vgamma9Vdelta2 T cell as new potential immunotherapy drug for solid tumor: a case study for cholangiocarcinoma. J. Immunother. Cancer 7, 36 (2019).
Hayes, S. M., Shores, E. W. & Love, P. E. An architectural perspective on signaling by the pre-, alphabeta and gammadelta T cell receptors. Immunol. Rev. 191, 28–37 (2003).
Rigau, M. et al. Butyrophilin 2A1 is essential for phosphoantigen reactivity by gammadelta T cells. Science 367, eaay5516 (2020).
Bertin, S. et al. The ion channel TRPV1 regulates the activation and proinflammatory properties of CD4(+) T cells. Nat. Immunol. 15, 1055–1063 (2014).
Guggino, W. B. & Stanton, B. A. New insights into cystic fibrosis: molecular switches that regulate CFTR. Nat. Rev. Mol. Cell Biol. 7, 426–436 (2006).
Roumier, A. et al. The membrane-microfilament linker ezrin is involved in the formation of the immunological synapse and in T cell activation. Immunity 15, 715–728 (2001).
Calabia-Linares, C. et al. Endosomal clathrin drives actin accumulation at the immunological synapse. J. Cell Sci. 124(Pt 5), 820–830 (2011).
Roncagalli, R. et al. Quantitative proteomics analysis of signalosome dynamics in primary T cells identifies the surface receptor CD6 as a Lat adaptor-independent TCR signaling hub. Nat. Immunol. 15, 384–392 (2014).
Chakraborty, A. K. & Weiss, A. Insights into the initiation of TCR signaling. Nat. Immunol. 15, 798–807 (2014).
Kouakanou, L. et al. Vitamin C promotes the proliferation and effector functions of human gammadelta T cells. Cell Mol. Immunol. 17, 462–473 (2019).
Carding, S. R. & Egan, P. J. Gammadelta T cells: functional plasticity and heterogeneity. Nat. Rev. Immunol. 2, 336–345 (2002).
Yin, Z. et al. Dominance of IL-12 over IL-4 in gamma delta T cell differentiation leads to default production of IFN-gamma: failure to down-regulate IL-12 receptor beta 2-chain expression. J. Immunol. 164, 3056–3064 (2000).
Yin, Z. et al. T-Bet expression and failure of GATA-3 cross-regulation lead to default production of IFN-gamma by gammadelta T cells. J. Immunol. 168, 1566–1571 (2002).
Sun, G. et al. gammadelta T cells provide the early source of IFN-gamma to aggravate lesions in spinal cord injury. J. Exp. Med. 215, 521–535 (2018).
Yang, Q. et al. Roles of mTORC1 and mTORC2 in controlling gammadelta T1 and gammadelta T17 differentiation and function. Cell Death Differ. 27, 2248–2262 (2020).
Ponzetto, A., Holton, J. & Lucia, U. Cancer risk in patients with cystic fibrosis. Gastroenterology 154, 2282–2283 (2018).
Liu, M. et al. Treatment of human T-cell acute lymphoblastic leukemia cells with CFTR inhibitor CFTRinh-172. Leuk. Res. 86, 106225 (2019).
Yamada, A. et al. Risk of gastrointestinal cancers in patients with cystic fibrosis: a systematic review and meta-analysis. Lancet Oncol. 19, 758–767 (2018).
Abraham, J. M. & Taylor, C. J. Cystic fibrosis & disorders of the large intestine: DIOS, constipation, and colorectal cancer. J. Cyst. Fibros. 16(Suppl 2), S40–S49 (2017).
Cao, G. et al. mTOR inhibition potentiates cytotoxicity of Vgamma4 gammadelta T cells via up-regulating NKG2D and TNF-alpha. J. Leukoc. Biol. 100, 1181–1189 (2016).
McEwen, G. D. et al. Subcellular spectroscopic markers, topography and nanomechanics of human lung cancer and breast cancer cells examined by combined confocal Raman microspectroscopy and atomic force microscopy. Analyst 138, 787–797 (2013).
Acknowledgements
This work was supported by grants from the National Natural Science Foundation of China (31420103901 to Z.Y., 31830021 to Z.Y., 31970830 to J.H., 81702876 to X.L., 31500734 to Y.D., and 31700753 to G.C.), grants from the Guangzhou Municipal Science and Technology Bureau (201904010090 to J.H. and 201906010085 to X.L.), and a grant from the Health Commission of Guangdong Province (A2019520 to J.H.).
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Y.D., G.L., and M.X. designed and performed experiments and data analysis. X.Q. and J.T. performed the patch clamp experiment and data analysis. Z.J., Q.Y., M.D., and Z.Lei helped with in vitro cell expansion. Y.H. performed the AFM experiment. Z.Li helped with data analysis and drawing of the proposed model. Z.Liu and Q.W. helped with animal breeding. X.L., G.C., W.K.Z., P.H., L.Z., and R.A.F. contributed to data analysis. Z.Y. and J.H. designed the research and wrote the manuscript.
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Duan, Y., Li, G., Xu, M. et al. CFTR is a negative regulator of γδ T cell IFN-γ production and antitumor immunity. Cell Mol Immunol 18, 1934–1944 (2021). https://doi.org/10.1038/s41423-020-0499-3
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DOI: https://doi.org/10.1038/s41423-020-0499-3