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    Topic review

    Stability of CD39+CD103+CD8+ T Cells

    Subjects: Oncology
    View times: 113
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    Definition

    Tumor-infiltrating CD8+ T cells (TIL) are of the utmost importance in anti-tumor immunity. CD103 defines tumor-resident memory T cells (TRM cells) associated with improved survival and response to immune checkpoint blockade (ICB) across human tumors. Co-expression of CD39 and CD103 marks tumor-specific TRM with enhanced cytolytic potential, suggesting that CD39+CD103+ TRM could be a suitable biomarker for immunotherapy. However, little is known about the transcriptional activity of TRM cells in situ. We analyzed CD39+CD103+ TRM cells sorted from human high-grade endometrial cancers (n = 3) using mRNA sequencing. Cells remained untreated or were incubated with PMA/ionomycin (activation), actinomycin D (a platinum-like chemotherapeutic that inhibits transcription), or a combination of the two. Resting CD39+CD103+ TRM cells were transcriptionally active and expressed a characteristic TRM signature. Activated CD39+CD103+ TRM cells differentially expressed PLEK, TWNK, and FOS, and cytokine genes IFNG, TNF, IL2, CSF2 (GM-CSF), and IL21. Findings were confirmed using qPCR and cytokine production was validated by flow cytometry of cytotoxic TIL. We studied transcript stability and found that PMA-responsive genes and mitochondrial genes were particularly stable.

    In conclusion, CD39+CD103+ TRM cells are transcriptionally active TRM cells with a polyfunctional, reactivation-responsive repertoire. Secondly, we hypothesize that differential regulation of transcript stability potentiates rapid responses upon TRM reactivation in tumors.

    1. Introduction

    The influence of tumor-infiltrating lymphocytes (TIL) on cancer prognosis is widely recognized, and TIL are studied in a wide variety of solid tumors. The composition of the immune infiltrate is of the utmost importance, as the immune architecture mainly determines whether the balance tips towards an anti-tumor or pro-tumor immune response [1]. CD103, the αE subunit of integrin αEβ7, defines intra-epithelial resident memory T cells (TRM cells) with increased cytolytic potential, improved immune synapse formation, and increased tumor antigen sensitivity [2][3][4]. CD8+ T cells upregulate CD103 upon combined TCR stimulation and TGF-β signaling [5][6][7]. Even though TGF-β production is commonly attributed to dendritic cells and T regulatory cells, differentiated CD103+ TRM are also capable of self-producing activated TGF-β1 to maintain CD103 expression on their cell surface [8]. CD103+ TRM are associated with prolonged survival in many solid tumors [4][6][9][10][11][12][13], including endometrial cancer (EC). CD103+ TRM, both in tumor and non-tumor tissue, are marked by expression of PDCD1ITGAECXCR6, and SPRY1 in lung cancer [19]. Tumor-resident CD103+ TRM expressed a unique genotype compared to non-tumor CD103+ TRM, characterized by expression of ENTPD1 (CD39) [20]. Indeed, bystander, i.e., non-tumor specific T cells, lack CD39 expression [21]. CD39, also known as ectonucleoside triphosphate diphosphohydrolase 1 (ENTPD1), catalyzes the phosphohydrolysis of extracellular ATP and ADP to eventually synthesize immunosuppressive adenosine. CD39 is upregulated on activated T cells [22], regulates T cell activation and polarization, and is considered an immunosuppressive marker associated with T cell exhaustion [23][24]. CD39 has therefore been put forth as an immunoregulatory checkpoint and a new therapeutic target in cancer [25]. A further specification of TRM cells in endometrial cancer might therefore be relevant, as CD39 and CD103 co-expression identifies tumor-resident, clonally expanded, tumor antigen-specific T cells with superior cytolytic capacity [19][20]. Moreover, tumor-resident CD103+ TRM differentially express immune checkpoints such as CTLA4, TIM3, LAG3, and TIGIT, indicating T cell exhaustion due to excess antigen stimulation [9][18][19][26]. In line with this, CD103+ TRM are linked to response to immune checkpoint blockade (ICB) [27][28]. Thus, it can be hypothesized that patients with both a sufficient number of TRM cells and a specific subtype of TRM cells are likely to respond to immunotherapy/immune checkpoint blockade. However, little is known about transcriptional activity of TRM cells in situ.

    2. Brief Summary

    We studied the transcriptional profile of high-grade endometrial cancer CD39+CD103+ TRM cells in situ, after T cell activation, and after transcriptional inhibition with actinomycin D in order to elucidate core elements necessary for successful T cell reactivation. Moreover, we studied the immune profile of TIL in the context of pretreatment with actinomycin D, a platinum-like chemotherapeutic. Resting CD39+CD103+ TRM cells were transcriptionally active and expressed a characteristic tissue-resident transcriptional profile, including several immune checkpoints. Upon activation, TRM cells upregulated markers of T cell activation, cytolytic activity, and cytokine production. Activated CD39+CD103+ TRM cells differentially expressed PLEK, TWNK, and FOS, and cytokine genes IFNG, TNF, IL2, CSF2(GM-CSF), and IL21. Secondly, we studied transcript stability and found that PMA-responsive immune genes and mitochondrial genes were particularly stable. 

    Taken together, our findings showed that CD39+CD103+ TRM cells in high-grade endometrial cancer are polyfunctional T cells with a reactivation-responsive repertoire, despite their exhausted phenotype. Secondly, CD39+CD103+ TRM showed increased transcript stability of PMA-responsive and mitochondrial genes, which may potentiate rapid responses upon TRM cell reactivation in tumors. Thirdly, TRM cells seem to incorporate activation-induced negative feedback mechanisms that halt TRM activation, and targeting these may be of interest in optimizing response to immune checkpoint blockade.

    This entry is adapted from 10.3390/ijms21113770

    References

    1. Barnes, T.A.; Amir, E. HYPE or HOPE: The prognostic value of infiltrating immune cells in cancer. Br. J. Cancer 2017.
    2. Franciszkiewicz, K.; Le Floc’H, A.; Boutet, M.; Vergnon, I.; Schmitt, A.; Mami-Chouaib, F. CD103 or LFA-1 engagement at the immune synapse between cytotoxic T cells and tumor cells promotes maturation and regulates T-cell effector functions. Cancer Res. 2013, 73, 617–628.
    3. Le Floc’h, A.; Jalil, A.; Franciszkiewicz, K.; Validire, P.; Vergnon, I.; Mami-Chouaib, F. Minimal engagement of CD103 on cytotoxic T lymphocytes with an E-cadherin-Fc molecule triggers lytic granule polarization via a phospholipase Cγ-dependent pathway. Cancer Res. 2011, 71, 328–338.
    4. Komdeur, F.L.; Prins, T.M.; van de Wall, S.; Plat, A.; Wisman, G.B.A.; Hollema, H.; Daemen, T.; Church, D.N.; de Bruyn, M.; Nijman, H.W. CD103+ tumor-infiltrating lymphocytes are tumor-reactive intraepithelial CD8+ T cells associated with prognostic benefit and therapy response in cervical cancer. Oncoimmunology 2017.
    5. Mokrani, M.; Klibi, J.; Bluteau, D.; Bismuth, G.; Mami-Chouaib, F. Smad and NFAT Pathways Cooperate To Induce CD103 Expression in Human CD8 T Lymphocytes. J. Immunol. 2014, 192, 2471–2479.
    6. Komdeur, F.L.; Wouters, M.C.A.; Workel, H.H.; Tijans, A.M.; Terwindt, A.L.J.; Brunekreeft, K.L.; Plat, A.; Klip, H.G.; Eggink, F.A.; Leffers, N.; et al. CD103+ intraepithelial T cells in high-grade serous ovarian cancer are phenotypically diverse TCRαβ+ CD8αβ+ T cells that can be targeted for cancer immunotherapy. Oncotarget 2016.
    7. Franciszkiewicz, K.; Le Floc’h, A.; Jalil, A.; Vigant, F.; Robert, T.; Vergnon, I.; Mackiewicz, A.; Benihoud, K.; Validire, P.; Chouaib, S.; et al. Intratumoral induction of CD103 triggers tumor-specific CTL function and CCR5-dependent T-cell retention. Cancer Res. 2009, 69, 6249–6255.
    8. Abd Hamid, M.; Colin-York, H.; Khalid-Alham, N.; Browne, M.; Cerundolo, L.; Chen, J.L.; Yao, X.; Rosendo-Machado, S.; Waugh, C.; Maldonado-Perez, D.; et al. Self-Maintaining CD103+ Cancer-Specific T Cells Are Highly Energetic with Rapid Cytotoxic and Effector Responses. Cancer Immunol. Res. 2020.
    9. Ganesan, A.-P.; Clarke, J.; Wood, O.; Garrido-Martin, E.M.; Chee, S.J.; Mellows, T.; Samaniego-Castruita, D.; Singh, D.; Seumois, G.; Alzetani, A.; et al. Tissue-resident memory features are linked to the magnitude of cytotoxic T cell responses in human lung cancer. Nat. Immunol. 2017, 18, 940–950.
    10. Wang, B.; Wu, S.; Zeng, H.; Liu, Z.; Dong, W.; He, W.; Chen, X.; Dong, X.; Zheng, L.; Lin, T.; et al. CD103+ Tumor Infiltrating Lymphocytes Predict a Favorable Prognosis in Urothelial Cell Carcinoma of the Bladder. J. Urol. 2015, 194, 556–562.
    11. Djenidi, F.; Adam, J.; Goubar, A.; Durgeau, A.; Meurice, G.; de Montpreville, V.; Validire, P.; Besse, B.; Mami-Chouaib, F. CD8+CD103+ Tumor-Infiltrating Lymphocytes Are Tumor-Specific Tissue-Resident Memory T Cells and a Prognostic Factor for Survival in Lung Cancer Patients. J. Immunol. 2015, 194, 3475–3486.
    12. Webb, J.R.; Milne, K.; Watson, P.; DeLeeuw, R.J.; Nelson, B.H. Tumor-infiltrating lymphocytes expressing the tissue resident memory marker cd103 are associated with increased survival in high-grade serous ovarian cancer. Clin. Cancer Res. 2014, 20, 434–444.
    13. Workel, H.H.; Komdeur, F.L.; Wouters, M.C.A.; Plat, A.; Klip, H.G.; Eggink, F.A.; Wisman, G.B.A.; Arts, H.J.G.; Oonk, M.H.M.; Mourits, M.J.E.; et al. CD103 defines intraepithelial CD8+ PD1+ tumour-infiltrating lymphocytes of prognostic significance in endometrial adenocarcinoma. Eur. J. Cancer 2016, 60, 1–11.
    14. Soslow, R.A.; Tornos, C.; Park, K.J.; Malpica, A.; Matias-Guiu, X.; Oliva, E.; Parkash, V.; Carlson, J.; Glenn McCluggage, W.; Blake Gilks, C. Endometrial Carcinoma Diagnosis: Use of FIGO Grading and Genomic Subcategories in Clinical Practice: Recommendations of the International Society of Gynecological Pathologists. Int. J. Gynecol. Pathol. 2019.
    15. McAlpine, J.; Leon-Castillo, A.; Bosse, T. The rise of a novel classification system for endometrial carcinoma; integration of molecular subclasses. J. Pathol. 2018.
    16. Eggink, F.A.; Van Gool, I.C.; Leary, A.; Pollock, P.M.; Crosbie, E.J.; Mileshkin, L.; Jordanova, E.S.; Adam, J.; Freeman-Mills, L.; Church, D.N.; et al. Immunological profiling of molecularly classified high-risk endometrial cancers identifies POLE-mutant and microsatellite unstable carcinomas as candidates for checkpoint inhibition. Oncoimmunology 2016, 6, e1264565.
    17. Ramchander, N.C.; Ryan, N.A.J.; Walker, T.D.J.; Harries, L.; Bolton, J.; Bosse, T.; Evans, D.G.; Crosbie, E.J. Distinct Immunological Landscapes Characterize Inherited and Sporadic Mismatch Repair Deficient Endometrial Cancer. Front. Immunol. 2020.
    18. Workel, H.H.; Lubbers, J.M.; Arnold, R.; Prins, T.M.; van der Vlies, P.; de Lange, K.; Bosse, T.; van Gool, I.C.; Eggink, F.A.; Wouters, M.C.A.; et al. A Transcriptionally Distinct CXCL13+CD103+CD8+ T-cell Population Is Associated with B-cell Recruitment and Neoantigen Load in Human Cancer. Cancer Immunol. Res. 2019.
    19. Clarke, J.; Panwar, B.; Madrigal, A.; Singh, D.; Gujar, R.; Wood, O.; Chee, S.J.; Eschweiler, S.; King, E.V.; Awad, A.S.; et al. Single-cell transcriptomic analysis of tissue-resident memory T cells in human lung cancer. J. Exp. Med. 2019.
    20. Duhen, T.; Duhen, R.; Montler, R.; Moses, J.; Moudgil, T.; De Miranda, N.F.; Goodall, C.P.; Blair, T.C.; Fox, B.A.; McDermott, J.E.; et al. Co-expression of CD39 and CD103 identifies tumor-reactive CD8 T cells in human solid tumors. Nat. Commun. 2018.
    21. Simoni, Y.; Becht, E.; Fehlings, M.; Loh, C.Y.; Koo, S.L.; Teng, K.W.W.; Yeong, J.P.S.; Nahar, R.; Zhang, T.; Kared, H.; et al. Bystander CD8+ T cells are abundant and phenotypically distinct in human tumour infiltrates. Nature 2018.
    22. Raczkowski, F.; Rissiek, A.; Ricklefs, I.; Heiss, K.; Schumacher, V.; Wundenberg, K.; Haag, F.; Koch-Nolte, F.; Tolosa, E.; Mittrücker, H.W. Cd39 is upregulated during activation of mouse and human t cells and attenuates the immune response to listeria monocytogenes. PLoS ONE 2018.
    23. Canale, F.P.; Ramello, M.C.; Núñez, N.; Furlan, C.L.A.; Bossio, S.N.; Serrán, M.G.; Boari, J.T.; Del Castillo, A.; Ledesma, M.; Sedlik, C.; et al. CD39 expression defines cell exhaustion in tumor-infiltrating CD8+ T cells. Cancer Res. 2018.
    24. Takenaka, M.C.; Robson, S.; Quintana, F.J. Regulation of the T Cell Response by CD39. Trends Immunol. 2016.
    25. Allard, B.; Longhi, M.S.; Robson, S.C.; Stagg, J. The ectonucleotidases CD39 and CD73: Novel checkpoint inhibitor targets. Immunol. Rev. 2017.
    26. Savas, P.; Virassamy, B.; Ye, C.; Salim, A.; Mintoff, C.P.; Caramia, F.; Salgado, R.; Byrne, D.J.; Teo, Z.L.; Dushyanthen, S.; et al. Single-cell profiling of breast cancer T cells reveals a tissue-resident memory subset associated with improved prognosis. Nat. Med. 2018.
    27. Park, S.L.; Buzzai, A.; Rautela, J.; Hor, J.L.; Hochheiser, K.; Effern, M.; McBain, N.; Wagner, T.; Edwards, J.; McConville, R.; et al. Tissue-resident memory CD8+ T cells promote melanoma–immune equilibrium in skin. Nature 2019.
    28. Edwards, J.; Wilmott, J.S.; Madore, J.; Gide, T.N.; Quek, C.; Tasker, A.; Ferguson, A.; Chen, J.; Hewavisenti, R.; Hersey, P.; et al. CD103+ tumor-resident CD8+ T cells are associated with improved survival in immunotherapy-naïve melanoma patients and expand significantly during anti-PD-1 treatment. Clin. Cancer Res. 2018
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