Human colorectal cancer: upregulation of the adaptor protein Rai in TILs leads to cell dysfunction by sustaining GSK-3 activation and PD-1 expression.
CRC
Molecular adaptor
PD-1
T cell
Tumor microenvironment
Journal
Cancer immunology, immunotherapy : CII
ISSN: 1432-0851
Titre abrégé: Cancer Immunol Immunother
Pays: Germany
ID NLM: 8605732
Informations de publication
Date de publication:
04 Jan 2024
04 Jan 2024
Historique:
received:
21
07
2023
accepted:
12
12
2023
medline:
4
1
2024
pubmed:
4
1
2024
entrez:
4
1
2024
Statut:
epublish
Résumé
The immunosuppressive tumor microenvironment (TME) of colorectal cancer (CRC) is a major hurdle for immune checkpoint inhibitor-based therapies. Hence characterization of the signaling pathways driving T cell exhaustion within TME is a critical need for the discovery of novel therapeutic targets and the development of effective therapies. We previously showed that (i) the adaptor protein Rai is a negative regulator of T cell receptor signaling and T helper 1 (Th1)/Th17 cell differentiation; and (ii) Rai deficiency is implicated in the hyperactive phenotype of T cells in autoimmune diseases. The expression level of Rai was measured by qRT-PCR in paired peripheral blood T cells and T cells infiltrating tumor tissue and the normal adjacent tissue in CRC patients. The impact of hypoxia-inducible factor (HIF)-1α on Rai expression was evaluated in T cells exposed to hypoxia and by performing chromatin immunoprecipitation assays and RNA interference assays. The mechanism by which upregulation of Rai in T cells promotes T cell exhaustion were evaluated by flow cytometric, qRT-PCR and western blot analyses. We show that Rai is a novel HIF-1α-responsive gene that is upregulated in tumor infiltrating lymphocytes of CRC patients compared to patient-matched circulating T cells. Rai upregulation in T cells promoted Programmed cell Death protein (PD)-1 expression and impaired antigen-dependent degranulation of CD8 Our data identify Rai as a hitherto unknown regulator of the TME-induced exhausted phenotype of human T cells.
Sections du résumé
BACKGROUND
BACKGROUND
The immunosuppressive tumor microenvironment (TME) of colorectal cancer (CRC) is a major hurdle for immune checkpoint inhibitor-based therapies. Hence characterization of the signaling pathways driving T cell exhaustion within TME is a critical need for the discovery of novel therapeutic targets and the development of effective therapies. We previously showed that (i) the adaptor protein Rai is a negative regulator of T cell receptor signaling and T helper 1 (Th1)/Th17 cell differentiation; and (ii) Rai deficiency is implicated in the hyperactive phenotype of T cells in autoimmune diseases.
METHODS
METHODS
The expression level of Rai was measured by qRT-PCR in paired peripheral blood T cells and T cells infiltrating tumor tissue and the normal adjacent tissue in CRC patients. The impact of hypoxia-inducible factor (HIF)-1α on Rai expression was evaluated in T cells exposed to hypoxia and by performing chromatin immunoprecipitation assays and RNA interference assays. The mechanism by which upregulation of Rai in T cells promotes T cell exhaustion were evaluated by flow cytometric, qRT-PCR and western blot analyses.
RESULTS
RESULTS
We show that Rai is a novel HIF-1α-responsive gene that is upregulated in tumor infiltrating lymphocytes of CRC patients compared to patient-matched circulating T cells. Rai upregulation in T cells promoted Programmed cell Death protein (PD)-1 expression and impaired antigen-dependent degranulation of CD8
CONCLUSIONS
CONCLUSIONS
Our data identify Rai as a hitherto unknown regulator of the TME-induced exhausted phenotype of human T cells.
Identifiants
pubmed: 38175205
doi: 10.1007/s00262-023-03614-0
pii: 10.1007/s00262-023-03614-0
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
2Subventions
Organisme : ERC Synergy
ID : ERC 951329
Organisme : Fondazione AIRC per la ricerca sul cancro ETS
ID : IG 2017-ID. 20148
Informations de copyright
© 2023. The Author(s).
Références
Ciardiello F, Ciardiello D, Martini G, Napolitano S, Tabernero J, Cervantes A (2022) Clinical management of metastatic Colorectal cancer in the era of precision medicine. CA A Cancer J Clinicians 72(4):372–401
doi: 10.3322/caac.21728
Dekker E, Tanis PJ, Vleugels JLA, Kasi PM, Wallace MB (2019) Colorectal cancer. The Lancet 394(10207):1467–1480
doi: 10.1016/S0140-6736(19)32319-0
Ganesh K, Stadler ZK, Cercek A, Mendelsohn RB, Shia J, Segal NH et al (2019) Immunotherapy in Colorectal cancer: rationale, challenges and potential. Nat Rev Gastroenterol Hepatol 16(6):361–375
pubmed: 30886395
pmcid: 7295073
doi: 10.1038/s41575-019-0126-x
Le DT, Durham JN, Smith KN, Wang H, Bartlett BR, Aulakh LK et al (2017) Mismatch repair deficiency predicts response of solid tumors to PD-1 blockade. Science 357(6349):409–413
pubmed: 28596308
pmcid: 5576142
doi: 10.1126/science.aan6733
Schumacher TN, Schreiber RD (2015) Neoantigens in cancer immunotherapy. Science 348(6230):69–74
pubmed: 25838375
doi: 10.1126/science.aaa4971
Verma NK, Wong BHS, Poh ZS, Udayakumar A, Verma R, Goh RKJ et al (2022) Obstacles for T-lymphocytes in the tumour microenvironment: therapeutic challenges, advances and opportunities beyond immune checkpoint. eBioMedicine 83:104216
pubmed: 35986950
pmcid: 9403334
doi: 10.1016/j.ebiom.2022.104216
Thommen DS, Schumacher TN (2018) T cell dysfunction in Cancer. Cancer Cell 33(4):547–562
pubmed: 29634943
pmcid: 7116508
doi: 10.1016/j.ccell.2018.03.012
Ahmadzadeh M, Johnson LA, Heemskerk B, Wunderlich JR, Dudley ME, White DE et al (2009) Tumor antigen–specific CD8 T cells infiltrating the Tumor express high levels of PD-1 and are functionally impaired. Blood 114(8):1537–1544
pubmed: 19423728
pmcid: 2927090
doi: 10.1182/blood-2008-12-195792
Scharping NE, Rivadeneira DB, Menk AV, Vignali PDA, Ford BR, Rittenhouse NL et al (2021) Mitochondrial stress induced by continuous stimulation under hypoxia rapidly drives T cell exhaustion. Nat Immunol 22(2):205–215
pubmed: 33398183
pmcid: 7971090
doi: 10.1038/s41590-020-00834-9
Zhang Y, Ertl HCJ (2016) Starved and asphyxiated: how can CD8 + T cells within a Tumor Microenvironment prevent Tumor Progression. Front Immunol. https://doi.org/10.3389/fimmu.2016.00032
doi: 10.3389/fimmu.2016.00032
pubmed: 28138326
pmcid: 5183739
Doedens AL, Phan AT, Stradner MH, Fujimoto JK, Nguyen JV, Yang E et al (2013) Hypoxia-inducible factors enhance the effector responses of CD8 + T cells to persistent antigen. Nat Immunol 14(11):1173–1182
pubmed: 24076634
pmcid: 3977965
doi: 10.1038/ni.2714
Bailey CM, Liu Y, Liu M, Du X, Devenport M, Zheng P et al (2022) Targeting HIF-1α abrogates PD-L1–mediated immune evasion in Tumor microenvironment but promotes tolerance in normal tissues. J Clin Invest 132(9):e150846
pubmed: 35239514
pmcid: 9057613
doi: 10.1172/JCI150846
Pietrobon V, Marincola FM (2021) Hypoxia and the phenomenon of immune exclusion. J Transl Med 19(1):9
pubmed: 33407613
pmcid: 7788724
doi: 10.1186/s12967-020-02667-4
Jayaprakash P, Ai M, Liu A, Budhani P, Bartkowiak T, Sheng J et al (2018) Targeted hypoxia reduction restores T cell infiltration and sensitizes Prostate cancer to immunotherapy. J Clin Invest 128(11):5137–5149
pubmed: 30188869
pmcid: 6205399
doi: 10.1172/JCI96268
Taylor A, Harker JA, Chanthong K, Stevenson PG, Zuniga EI, Rudd CE (2016) Glycogen synthase kinase 3 inactivation drives T-bet-mediated downregulation of co-receptor PD-1 to enhance CD8 + cytolytic T cell responses. Immunity 44(2):274–286
pubmed: 26885856
pmcid: 4760122
doi: 10.1016/j.immuni.2016.01.018
Steele L, Mannion AJ, Shaw G, Maclennan KA, Cook GP, Rudd CE et al (2021) Non-redundant activity of GSK-3α and GSK-3β in T cell-mediated Tumor rejection. iScience 24(6):102555
pubmed: 34142056
pmcid: 8188550
doi: 10.1016/j.isci.2021.102555
Ohteki T, Parsons M, Zakarian A, Jones RG, Nguyen LT, Woodgett JR et al (2000) Negative regulation of T cell proliferation and interleukin 2 production by the serine threonine kinase Gsk-3. J Exp Med 192(1):99–104
pubmed: 10880530
pmcid: 1887707
doi: 10.1084/jem.192.1.99
Taylor A, Rothstein D, Rudd CE (2018) Small-molecule inhibition of PD-1 transcription is an effective alternative to antibody blockade in Cancer Therapy. Cancer Res 78(3):706–717
pubmed: 29055015
doi: 10.1158/0008-5472.CAN-17-0491
Rudd CE, Chanthong K, Taylor A (2020) Small molecule inhibition of GSK-3 specifically inhibits the transcription of inhibitory co-receptor LAG-3 for enhanced anti-tumor immunity. Cell Rep 30(7):2075–2082e4
pubmed: 32075731
doi: 10.1016/j.celrep.2020.01.076
Ferro M, Savino MT, Ortensi B, Finetti F, Genovese L, Masi G et al (2011) The Shc family protein adaptor, Rai, negatively regulates T cell antigen receptor signaling by inhibiting ZAP-70 recruitment and activation. Alberola-Ila J, editor. PLoS ONE 6(12):e29899
Savino MT, Ortensi B, Ferro M, Ulivieri C, Fanigliulo D, Paccagnini E et al (2009) Rai acts as a negative regulator of autoimmunity by inhibiting antigen receptor signaling and lymphocyte activation. J Immunol 182(1):301–308
pubmed: 19109161
doi: 10.4049/jimmunol.182.1.301
Savino MT, Ulivieri C, Emmi G, Prisco D, De Falco G, Ortensi B et al (2013) The shc family protein adaptor, Rai, acts as a negative regulator of Th17 and Th1 cell development. J Leukoc Biol 93(4):549–559
pubmed: 23345394
doi: 10.1189/jlb.0712331
Chicaybam L, Sodre AL, Curzio BA, Bonamino MH (2013) An efficient low cost method for gene transfer to T lymphocytes. Fang D, editor. PLoS ONE 8(3):e60298
Monaci S, Aldinucci C, Rossi D, Giuntini G, Filippi I, Ulivieri C et al (2020) Hypoxia shapes autophagy in LPS-activated dendritic cells. Front Immunol 11:573646
pubmed: 33329536
pmcid: 7734254
doi: 10.3389/fimmu.2020.573646
Staller P, Sulitkova J, Lisztwan J, Moch H, Oakeley EJ, Krek W (2003) Chemokine receptor CXCR4 downregulated by Von Hippel–Lindau tumour suppressor pVHL. Nature 425(6955):307–311
pubmed: 13679920
doi: 10.1038/nature01874
Filippi I, Morena E, Aldinucci C, Carraro F, Sozzani S, Naldini A (2014) Short-term Hypoxia enhances the migratory capability of dendritic cell through HIF-1α and PI3K/Akt pathway: dendritic cell migration under hypoxia. J Cell Physiol 229(12):2067–2076
pubmed: 24818793
doi: 10.1002/jcp.24666
Xu K, Zhan Y, Yuan Z, Qiu Y, Wang H, Fan G et al (2019) Hypoxia induces drug resistance in colorectal cancer through the HIF-1α/miR-338-5p/IL-6 feedback loop. Mol Ther 27(10):1810–1824
pubmed: 31208913
pmcid: 6822233
doi: 10.1016/j.ymthe.2019.05.017
McKeown SR (2014) Defining normoxia, physoxia and hypoxia in tumours—implications for treatment response. BJR 87(1035):20130676
pubmed: 24588669
pmcid: 4064601
doi: 10.1259/bjr.20130676
Palazon A, Goldrath AW, Nizet V, Johnson RS (2014) HIF transcription factors, inflammation, and immunity. Immunity 41(4):518–528
pubmed: 25367569
pmcid: 4346319
doi: 10.1016/j.immuni.2014.09.008
Golstein P, Griffiths GM (2018) An early history of T cell-mediated cytotoxicity. Nat Rev Immunol 18(8):527–535
pubmed: 29662120
doi: 10.1038/s41577-018-0009-3
Russo E, Gloria LD, Nannini G, Meoni G, Niccolai E, Ringressi MN et al (2023) From adenoma to CRC stages: the oral-gut microbiome axis as a source of potential microbial and metabolic biomarkers of malignancy. Neoplasia 40:100901
pubmed: 37058886
pmcid: 10130693
doi: 10.1016/j.neo.2023.100901
Nannini G, De Luca V, D’Ambrosio C, Scaloni A, Taddei A, Ringressi MN et al (2022) A comparative study of carbonic anhydrase activity in lymphocytes from colorectal cancer tissues and adjacent healthy counterparts. J Enzyme Inhib Med Chem 37(1):1651–1655
pubmed: 35695123
pmcid: 9225793
doi: 10.1080/14756366.2022.2085694
Niccolai E, Russo E, Baldi S, Ricci F, Nannini G, Pedone M et al (2021) Significant and conflicting correlation of IL-9 with prevotella and bacteroides in human colorectal cancer. Front Immunol 11:573158
pubmed: 33488574
pmcid: 7820867
doi: 10.3389/fimmu.2020.573158
Qiao Y, Jiang X, Li Y, Wang K, Chen R, Liu J et al (2023) Identification of a hypoxia-related gene prognostic signature in Colorectal cancer based on bulk and single-cell RNA-seq. Sci Rep 13(1):2503
pubmed: 36781976
pmcid: 9925779
doi: 10.1038/s41598-023-29718-2
Semenza GL (2014) Oxygen sensing, hypoxia-inducible factors, and disease pathophysiology. Annu Rev Pathol Mech Dis 9(1):47–71
doi: 10.1146/annurev-pathol-012513-104720
Colegio OR, Chu NQ, Szabo AL, Chu T, Rhebergen AM, Jairam V et al (2014) Functional polarization of tumour-associated macrophages by tumour-derived lactic acid. Nature 513(7519):559–563
pubmed: 25043024
pmcid: 4301845
doi: 10.1038/nature13490
Fischbeck AJ, Ruehland S, Ettinger A, Paetzold K, Masouris I, Noessner E et al (2020) Tumor lactic acidosis: protecting tumor by inhibiting cytotoxic activity through motility arrest and bioenergetic silencing. Front Oncol 10:589434
pubmed: 33364193
pmcid: 7753121
doi: 10.3389/fonc.2020.589434
Wei TT, Lin YT, Tang SP, Luo CK, Tsai CT, Shun CT et al (2020) Metabolic targeting of HIF-1α potentiates the therapeutic efficacy of oxaliplatin in colorectal cancer. Oncogene 39(2):414–427
pubmed: 31477841
doi: 10.1038/s41388-019-0999-8
Niu H, Wang H (2023) TOX regulates T lymphocytes differentiation and its function in tumor. Front Immunol 14:990419
pubmed: 36969216
pmcid: 10035881
doi: 10.3389/fimmu.2023.990419
Kim CG, Jang M, Kim Y, Leem G, Kim KH, Lee H et al (2019) VEGF-A drives TOX-dependent T cell exhaustion in anti–PD-1–resistant microsatellite stable colorectal cancers. Sci Immunol 4(41):eaay0555
pubmed: 31704735
doi: 10.1126/sciimmunol.aay0555
Shurin MR, Umansky V (2022) Cross-talk between HIF and PD-1/PD-L1 pathways in carcinogenesis and therapy. J Clin Invest 132(9):e159473
pubmed: 35499071
pmcid: 9057611
doi: 10.1172/JCI159473
Feldhoff LM, Rueda CM, Moreno-Fernandez ME, Sauer J, Jackson CM, Chougnet CA et al (2017) IL-1β induced HIF-1α inhibits the differentiation of human FOXP3 + T cells. Sci Rep 7(1):465
pubmed: 28352109
pmcid: 5428734
doi: 10.1038/s41598-017-00508-x
Clambey ET, McNamee EN, Westrich JA, Glover LE, Campbell EL, Jedlicka P et al (2012) Hypoxia-inducible factor-1 alpha–dependent induction of FoxP3 drives regulatory T-cell abundance and function during inflammatory hypoxia of the mucosa. Proc Natl Acad Sci USA 109(41)
Liikanen I, Lauhan C, Quon S, Omilusik K, Phan AT, Bartrolí LB et al (2021) Hypoxia-inducible factor activity promotes antitumor effector function and tissue residency by CD8 + T cells. J Clin Invest 131(7):e143729
pubmed: 33792560
pmcid: 8011896
doi: 10.1172/JCI143729
Bannoud N, Dalotto-Moreno T, Kindgard L, García PA, Blidner AG, Mariño KV et al (2021) Hypoxia supports differentiation of terminally exhausted CD8 T cells. Front Immunol 12:660944
pubmed: 34025660
pmcid: 8137905
doi: 10.3389/fimmu.2021.660944
Scharping N, Delgoffe G (2016) Tumor microenvironment metabolism: a new checkpoint for anti-tumor immunity. Vaccines 4(4):46
pubmed: 27929420
pmcid: 5192366
doi: 10.3390/vaccines4040046
Elliot TAE, Jennings EK, Lecky DAJ, Thawait N, Flores-Langarica A, Copland A et al (2021) Antigen and checkpoint receptor engagement recalibrates T cell receptor signal strength. Immunity 54(11):2481–2496e6
pubmed: 34534438
pmcid: 8585507
doi: 10.1016/j.immuni.2021.08.020