Myeloid-derived Suppressor Cells Activate Liver Natural Killer Cells in a Murine Model in Uveal Melanoma

Abstract

Objective

Elevated myeloid-derived suppressor cells (MDSCs) in many malignancies are associated with the increased risk for metastases and poor prognosis. Therefore, a mouse model of intraocular melanoma was established to explore how MDSCs influence liver metastases.

Methods

In this study, murine B16LS melanoma cells were transplanted into the posterior compartment (PC) of the eye of C57BL/6 mice. Leucocytes from the liver of naive mice and mice bearing melanoma liver metastasis were isolated using isotonic Percoll centrifugation, examined by flow cytometry for their expression of Gr1, CD11b, F4/80, RAE-1, and Mult-1, and further isolated for MDSCs and natural killer (NK) cells. The effects of MDSCs on NK cells were tested by coculturing and assessing the ability of NK cells to produce interferon-gamma (IFN-γ) by ELISA and NK cell cytotoxicity by ³H-thymidine incorporation assay. The impact of IFN-γ on liver metastases was examined via selectively depleting IFN-γ in vivo.

Results

The results showed that mice with liver metastases had increased levels of CD11b⁺Gr1⁺F4/80⁺ as well as CD11b⁺Gr1⁺F4/80⁻ MDSCs. MDSCs significantly enhanced the generation of IFN-γ together with the cytotoxicity of the NK cells. Furthermore, these effects were cell-cell contact-dependent. Although IFN-γ was not of a toxic nature to the melanoma cells, it profoundly inhibited B16LS cell proliferation. Depleting IFN-γ in vivo led to increased liver metastases.

Conclusion

All these findings first revealed that MDSCs accumulated in liver metastasis of intraocular melanoma could activate the NK cells to produce an effective anti-tumor immune response. Thus, the MDSCs’ performance in different tumor models would need more investigation to boost current immunotherapy modalities.

Keywords

myeloid-derived suppressor cells / natural killer cells / IFN-γ / liver metastases

References

[1] GrossniklausHE. Progression of ocular melanoma metastasis to the liver: the 2012 Zimmerman lecture. JAMA Ophthalmol, 2013, 131(4): 462-469

[2] JakošT, PišlarA, JewettA, et al.. Myeloid-Derived Suppressor Cells Hamper Natural Killer Cell Activity in Cancer: Role of Peptidases. Crit Rev Immunol, 2021, 41(2): 77-99

[3] JonesNM, YangH, ZhangQ, et al.. Natural killer cells and pigment epithelial-derived factor control the infiltrative and nodular growth of hepatic metastases in an Orthotopic murine model of ocular melanoma. BMC Cancer, 2019, 19(1): 484

[4] GuiaS, FenisA, VivierE, et al.. Activating and inhibitory receptors expressed on innate lymphoid cells. Semin Immunopathol, 2018, 40(4): 331-341

[5] MalhotraA, ShankerA. NK cells: immune crosstalk and therapeutic implications. Immunotherapy, 2011, 3(10): 1143-1166

[6] ShimasakiN, JainA, CampanaD. NK cells for cancer immunotherapy. Nat Rev Drug Discov, 2020, 19(3): 200-218

[7] TuminoN, Di PaceAL, BesiF, et al.. Interaction Between MDSC and NK Cells in Solid and Hematological Malignancies: Impact on HSCT. Front Immunol, 2021, 12: 638841

[8] HegdeS, LeaderAM, MeradM. MDSC: Markers, development, states, and unaddressed complexity. Immunity, 2021, 54(5): 875-884

[9] VanhaverC, van der BruggenP, BrugerAM. MDSC in Mice and Men: Mechanisms of Immunosuppression in Cancer. J Clin Med, 2021, 10(13): 2872

[10] NauschN, GalaniIE, SchleckerE, et al.. Mononuclear myeloid-derived “suppressor” cells express RAE-1 and activate natural killer cells. Blood, 2008, 112(10): 4080-4089

[11] ZalfaC, PaustS. Natural Killer Cell Interactions With Myeloid Derived Suppressor Cells in the Tumor Microenvironment and Implications for Cancer Immunotherapy. Front Immunol, 2021, 12: 633205

[12] LiuC, YuS, KappesJ, et al.. Expansion of spleen myeloid suppressor cells represses NK cell cytotoxicity in tumor-bearing host. Blood, 2007, 109(10): 4336-4342

[13] RuscianoD, LorenzoniP, BurgerMM. Paracrine growth response as a major determinant in liver-specific colonization by in vivo selected B16 murine melanoma cells. Invasion Metastasis, 1993, 13(4): 212-224

[14] DiazCE, RuscianoD, DithmarS, et al.. B16LS9 melanoma cells spread to the liver from the murine ocular posterior compartment (PC). Curr Eye Res, 1999, 18(2): 125-129

[15] YangW, LiH, MayhewE, et al.. NKT cell exacerbation of liver metastases arising from melanomas transplanted into either the eyes or spleens of mice. Invest Ophthalmol Vis Sci, 2011, 52(6): 3094-3102

[16] WuZ, ZhangH, WuM, et al.. Targeting the NKG2D/NKG2D-L axis in acute myeloid leukemia. Biomed Pharmacother, 2021, 137: 111299

[17] ZhangQF, YinWW, XiaY, et al.. Liver-infiltrating CD11b-CD27- NK subsets account for NK-cell dysfunction in patients with hepatocellular carcinoma and are associated with tumor progression. Cell Mol Immunol, 2017, 14(10): 819-829

[18] ZhuangL, FultonRJ, RettmanP, et al.. Activity of IL-12/15/18 primed natural killer cells against hepatocellular carcinoma. Hepatol Int, 2019, 13(1): 75-83

[19] NiC, WuP, ZhuX, et al.. IFN-γ selectively exerts proapoptotic effects on tumor-initiating label-retaining colon cancer cells. Cancer Lett, 2013, 336(1): 174-184

[20] SubleskiJJ, HallVL, BackTC, et al.. Enhanced antitumor response by divergent modulation of natural killer and natural killer T cells in the liver. Cancer Res, 2006, 66(22): 11005-11012

[21] FranksSE, WolfsonB, HodgeJW. Natural Born Killers: NK Cells in Cancer Therapy. Cancers (Basel), 2020, 12(8): 2131

[22] OhlK, TenbrockK. Reactive Oxygen Species as Regulators of MDSC-Mediated Immune Suppression. Front Immunol, 2018, 9: 2499

[23] ChaiE, ZhangL, LiC. LOX-1+ PMN-MDSC enhances immune suppression which promotes glioblastoma multiforme progression. Cancer Manag Res, 2019, 11: 7307-7315

[24] TesiRJ. MDSC; the Most Important Cell You Have Never Heard Of. Trends Pharmacol Sci, 2019, 40(1): 4-7

[25] MeyerC, CagnonL, Costa-NunesCM, et al.. Frequencies of circulating MDSC correlate with clinical outcome of melanoma patients treated with ipilimumab. Cancer Immunol Immunother, 2014, 63(3): 247-257

[26] MarvelD, GabrilovichDI. Myeloid-derived suppressor cells in the tumor microenvironment: expect the unexpected. J Clin Invest, 2015, 125(9): 3356-3364

[27] NajjarYG, FinkeJH. Clinical perspectives on targeting of myeloid derived suppressor cells in the treatment of cancer. Front Oncol, 2013, 3: 49

[28] LiH, HanY, GuoQ, et al.. Cancer-expanded myeloid-derived suppressor cells induce anergy of NK cells through membrane-bound TGF-beta 1. J Immunol, 2009, 182(1): 240-249

[29] AngelicolaS, RuzziF, LanduzziL, et al.. IFN-γ and CD38 in Hyperprogressive Cancer Development. Cancers (Basel), 2021, 13(2): 309

[30] SarmadiP, TunaliG, Esendagli-YilmazG, et al.. CRAM-A indicates IFN-γ-associated inflammatory response in breast cancer. Mol Immunol, 2015, 68(2PtC): 692-698

[31] ImaiD, YoshizumiT, OkanoS, et al.. IFN-γ Promotes Epithelial-Mesenchymal Transition and the Expression of PD-L1 in Pancreatic Cancer. J Surg Res, 2019, 240: 115-123