A YAP/TAZ-CD54 axis is required for CXCR2⁻CD44⁻ tumor-specific neutrophils to suppress gastric cancer

Abstract

As an important part of tumor microenvironment, neutrophils are poorly understood due to their spatiotemporal heterogeneity in tumorigenesis. Here we defined, at single-cell resolution, CD44⁻CXCR2⁻ neutrophils as tumor-specific neutrophils (tsNeus) in both mouse and human gastric cancer (GC). We uncovered a Hippo regulon in neutrophils with unique YAP signature genes (e.g., ICAM1, CD14, EGR1) distinct from those identified in epithelial and/or cancer cells. Importantly, knockout of YAP/TAZ in neutrophils impaired their differentiation into CD54⁺ tsNeus and reduced their antitumor activity, leading to accelerated GC progression. Moreover, the relative amounts of CD54⁺ tsNeus were found to be negatively associated with GC progression and positively associated with patient survival. Interestingly, GC patients receiving neoadjuvant chemotherapy had increased numbers of CD54⁺ tsNeus. Furthermore, pharmacologically enhancing YAP activity selectively activated neutrophils to suppress refractory GC, with no significant inflammation-related side effects. Thus, our work characterized tumor-specific neutrophils in GC and revealed an essential role of YAP/TAZ-CD54 axis in tsNeus, opening a new possibility to develop neutrophil-based antitumor therapeutics.

Keywords

neutrophils / YAP / TAZ / gastric cancer

References

[1] Aibar S, Gonzalez-Blas CB, Moerman T et al. SCENIC: single-cell regulatory network inference and clustering. Nat Methods 2017;14:1083–1086.

[2] Albrengues J, Shields MA, Ng D et al. Neutrophil extracellular traps produced during inflammation awaken dormant cancer cells in mice. Science 2018;361:eaao4227.

[3] Ballesteros I, Rubio-Ponce A, Genua M et al. Co-option of neutrophil fates by tissue environments. Cell 2020.

[4] Beinert T, Münzing S, Possinger K et al. Increased expression of the tetraspanins CD53 and CD63 on apoptotic human neutrophils. J Leukoc Biol 2000;67:369–373.

[5] Blaisdell A, Crequer A, Columbus D et al. Neutrophils oppose uterine epithelial carcinogenesis via debridement of hypoxic tumor cells. Cancer Cell 2015;28:785–799.

[6] Borregaard N. Neutrophils, from marrow to microbes. Immunity 2010;33:657–670.

[7] Borregaard, N, Sorensen OE, Theilgaard-Monch K. Neutrophil granules: a library of innate immunity proteins. Trends Immunol 2007;28:340–345.

[8] Cools-Lartigue J, Spicer J, McDonald B et al. Neutrophil extracellular traps sequester circulating tumor cells and promote metastasis. J Clin Invest 2013;123:3446–3458.

[9] Cowland JB, Borregaard N. Granulopoiesis and granules of human neutrophils. Immunol Rev 2016;273:11–28.

[10] Cui C, Chakraborty K, Tang XA et al. Neutrophil elastase selectively kills cancer cells and attenuates tumorigenesis. Cell 2021;184:3163–3177.

[11] Dinh HQ, Eggert T, Meyer MA et al. Coexpression of CD71 and CD117 identifies an early unipotent neutrophil progenitor population in human bone marrow. Immunity 2020; 53:319–334.

[12] Eash KJ, Greenbaum AM, Gopalan PK et al. CXCR2 and CXCR4 antagonistically regulate neutrophil trafficking from murine bone marrow. J Clin Invest 2010;120:2423–2431.

[13] Elliott ER, Van Ziffle JA, Scapini P et al. Deletion of Syk in neutrophils prevents immune complex arthritis. J Immunol 2011;187:4319–4330.

[14] Eruslanov EB, Bhojnagarwala PS, Quatromoni JG et al. Tumor-associated neutrophils stimulate T cell responses in early-stage human lung cancer. J Clin Invest 2014;124:5466–5480.

[15] Evrard M, Kwok IWH, Chong SZ et al. Developmental analysis of bone marrow neutrophils reveals populations specialized in expansion, trafficking, and effector functions. Immunity 2018a;48;364–379.

[16] Evrard M, Kwok IWH, Chong SZ et al. Developmental analysis of bone marrow neutrophils reveals populations specialized in expansion, trafficking, and effector functions. Immunity 2018b;48:364–379.

[17] Fan F, He Z, Kong LL et al. Pharmacological targeting of kinases MST1 and MST2 augments tissue repair and regeneration. Sci Transl Med 2016;8:352ra108.

[18] Finisguerra V, Di Conza G, Di Matteo M et al. MET is required for the recruitment of anti-tumoural neutrophils. Nature 2015;522:349–353.

[19] Geng J, Yu S, Zhao H et al. The transcriptional coactivator TAZ regulates reciprocal differentiation of TH17 cells and Treg cells. Nat Immunol 2017;18:800–812.

[20] Gershkovitz M, Caspi Y, Fainsod-Levi T et al. TRPM2 Mediates Neutrophil Killing of Disseminated Tumor Cells. Cancer Res 2018;78:2680–2690.

[21] Granot Z, Henke E, Comen EA et al. Tumor entrained neutrophils inhibit seeding in the premetastatic lung. Cancer Cell 2011;20:300–314.

[22] Harvey KF, Zhang X, Thomas DM. The Hippo pathway and human cancer. Nat Rev Cancer 2013;13:246–257.

[23] Hock H, Hamblen MJ, Rooke HM et al. Intrinsic requirement for zinc finger transcription factor Gfi-1 in neutrophil differentiation. Immunity 2003;18:109–120.

[24] Huang B, Pan PY, Li Q et al. Gr-1+CD115+ immature myeloid suppressor cells mediate the development of tumor-induced T regulatory cells and T-cell anergy in tumor-bearing host. Cancer Res 2006;66:1123–1131.

[25] Jaillon S, Ponzetta A, Di Mitri D et al. Neutrophil diversity and plasticity in tumour progression and therapy. Nat Rev Cancer 2020;20:485–503.

[26] Jiao S, Wang H, Shi Z et al. A peptide mimicking VGLL4 function acts as a YAP antagonist therapy against gastric cancer. Cancer Cell 2014;25:166–180.

[27] Jiao S, Zhang Z, Li C et al. The kinase MST4 limits inflammatory responses through direct phosphorylation of the adaptor TRAF6. Nat Immunol 2015;16:246–257.

[28] Kolaczkowska E, Kubes P. Neutrophil recruitment and function in health and inflammation. Nat Rev Immunol 2013;13:159–175.

[29] Kwok I, Becht E, Xia Y et al. Combinatorial single-cell analyses of granulocyte- monocyte progenitor heterogeneity reveals an early uni-potent neutrophil progenitor. Immunity 2020;53:303–318.

[30] Liew PX, Kubes P. The neutrophil’s role during health and disease. Physiol Rev 2019;99:1223–1248.

[31] McDonald B, McAvoy EF, Lam F et al. Interaction of CD44 and hyaluronan is the dominant mechanism for neutrophil sequestration in inflamed liver sinusoids. J Exp Med 2008;205:915–927.

[32] Mensurado S, Rei M, Lança T et al. Tumor-associated neutrophils suppress pro-tumoral IL-17+ γδ T cells through induction of oxidative stress. PLoS Biol 2018;16:e2004990.

[33] Metkar S, Kim KS, Silver J et al. Differential expression of CD14-dependent and independent pathways for chemokine induction regulates neutrophil trafficking in infection. J Leukoc Biol 2012;92:389–396.

[34] Németh T, Sperandio M, Mócsai A. Neutrophils as emerging therapeutic targets. Nat Rev Drug Discovery 2020;19:253–275.

[35] Ng LG, Ostuni R, Hidalgo A. Heterogeneity of neutrophils. Nat Rev Immunol 2019;19:255–265.

[36] Ni X, Tao J, Barbi J et al. YAP is essential for treg-mediated suppression of antitumor immunity. Cancer Discov 2018;8:1026–1043.

[37] Nozawa H, Chiu C, Hanahan D. Infiltrating neutrophils mediate the initial angiogenic switch in a mouse model of multistage carcinogenesis. Proc Natl Acad Sci USA 2006;103:12493.

[38] Ostrand-Rosenberg S, Sinha P. Myeloid-derived suppressor cells: linking inflammation and cancer. J Immunol 2009;182:4499–4506.

[39] Pan D. The hippo signaling pathway in development and cancer. Dev Cell 2010;19:491–505.

[40] Ponzetta A, Carriero R, Carnevale S et al. Neutrophils driving unconventional t cells mediate resistance against murine sarcomas and selected human tumors. Cell 2019;178:346–360.

[41] Qiu X, Mao Q, Tang Y et al. Reversed graph embedding resolves complex single-cell trajectories. Nat Methods 2017;14:979–982.

[42] Ren X, Zhong G, Zhang Q et al. Reconstruction of cell spatial organization from single-cell RNA sequencing data based on ligand-receptor mediated self-assembly. Cell Res 2020;30:763–778.

[43] Shaul ME, Fridlender ZG. Tumour-associated neutrophils in patients with cancer. Nat Rev Clin Oncol 2019;16:601–620.

[44] Shojaei F, Singh M, Thompson JD et al. Role of Bv8 in neutrophil-dependent angiogenesis in a transgenic model of cancer progression. Proc Natl Acad Sci USA 2008;105:2640–2645.

[45] Singhal S, Bhojnagarwala PS, O’Brien S et al. Origin and role of a subset of tumor-associated neutrophils with antigen-presenting cell features in early-stage human lung cancer. Cancer Cell 2016;30:120–135.

[46] Spiegel A, Brooks MW, Houshyar S et al. Neutrophils suppress intraluminal NK cell-mediated tumor cell clearance and enhance extravasation of disseminated carcinoma cells. Cancer Discov 2016;6:630–649.

[47] Szczerba BM, Castro-Giner F, Vetter M et al. Neutrophils escort circulating tumour cells to enable cell cycle progression. Nature 2019;566:553–557.

[48] Tang Y, Fang G, Guo F et al. Selective inhibition of STRN3-containing PP2A phosphatase restores Hippo tumor-suppressor activity in gastric cancer. Cancer Cell 2020;38:115–128.

[49] van der Maaten L, Hinton G. Visualizing data using t-SNE. J Mach Learn Res 2008;9:2579–2605.

[50] Veglia F, Hashimoto A, Dweep H et al. Analysis of classical neutrophils and polymorphonuclear myeloid-derived suppressor cells in cancer patients and tumor-bearing mice. J Exp Med 2021;218.

[51] Wang TT, Zhao YL, Peng LS et al. Tumour-activated neutrophils in gastric cancer foster immune suppression and disease progression through GM-CSF-PD-L1 pathway. Gut 2017;66:1900–1911.

[52] Xie X, Shi Q, Wu P et al. Single-cell transcriptome profiling reveals neutrophil heterogeneity in homeostasis and infection. Nat Immunol 2020a;21:1119–1133.

[53] Xie X, Shi Q, Wu P et al. Single-cell transcriptome profiling reveals neutrophil heterogeneity in homeostasis and infection. Nat Immunol 2020b;21:1119–1133.

[54] Yamanaka R, Barlow C, Lekstrom-Himes J et al. Impaired granulopoiesis, myelodysplasia, and early lethality in CCAAT/enhancer binding protein epsilon-deficient mice. Proc Natl Acad Sci USA 1997;94:13187–13192.

[55] Yang L, Liu Q, Zhang X et al. DNA of neutrophil extracellular traps promotes cancer metastasis via CCDC25. Nature 2020;583:133–138.

[56] Youn JI, Collazo M, Shalova IN et al. Characterization of the nature of granulocytic myeloid-derived suppressor cells in tumor-bearing mice. J Leukoc Biol 2012;91:167–181.

[57] Yu FX, Meng Z, Plouffe SW et al. Hippo pathway regulation of gastro-intestinal tissues. Annu Rev Physiol 2015;77:201–227.

[58] Zanconato F, Forcato M, Battilana G et al. Genome-wide association between YAP/TAZ/TEAD and AP-1 at enhancers drives oncogenic growth. Nat Cell Biol 2015;17:1218–1227.

[59] Zhang H, Liu CY, Zha ZY et al. TEAD transcription factors mediate the function of TAZ in cell growth and epithelial-mesenchymal transition. J Biol Chem 2009;284:13355–13362.

[60] Zhao B, Tumaneng K, Guan KL. The Hippo pathway in organ size control, tissue regeneration and stem cell self-renewal. Nat Cell Biol 2011;13:877–883.

[61] Zhao B, Ye X, Yu J et al. TEAD mediates YAP-dependent gene induction and growth control. Genes Dev 2008;22:1962–1971.

[62] Zhu YP, Padgett L, Dinh HQ et al. Identification of an early unipotent neutrophil progenitor with pro-tumoral activity in mouse and human bone marrow. Cell Rep 2018;24:2329–2341.

[63] Zilionis R, Engblom C, Pfirschke C et al. Single-cell transcriptomics of human and mouse lung cancers reveals conserved myeloid populations across individuals and species. Immunity 2019;50:1317–1334.

RIGHTS & PERMISSIONS

The Author(s) 2022. Published by Oxford University Press on behalf of Higher Education Press.