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Gastric cancer mesenchymal stem cells via the CXCR2/HK2/PD-L1 pathway mediate immunosuppression

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Abstract

Background

Anti-PD-1 immunotherapy has emerged as an important therapeutic modality in advanced gastric cancer (GC). However, drug resistance frequently develops, limiting its effectiveness.

Methods

The role of gastric cancer mesenchymal stem cells (GCMSCs) in anti-PD-1 resistance was evaluated in vivo in NPGCD34+ or NCGPBMC xenograft mouse model. In addition, we investigated CD8+T cell infiltration and effector function by spectral cytometry and IHC. The effects of GCMSCs conditional medium (GCMSC-CM) on GC cell lines were characterized at the level of the proteome, secretome using western blot, and ELISA assays.

Results

We reported that GCMSCs mediated tolerance mechanisms contribute to tumor immunotherapy tolerance. GCMSC-CM attenuated the antitumor activity of PD-1 antibody and inhibited immune response in humanized mouse model. In GC cells under serum deprivation and hypoxia, GCMSC-CM promoted GC cells proliferation via upregulating PD-L1 expression. Mechanistically, GCMSC-derived IL-8 and AKT-mediated phosphorylation facilitated HK2 nuclear localization. Phosphorylated-HK2 promoted PD-L1 transcription by binding to HIF-1α. What is more, GCMSC-CM also induced lactate overproduction in GC cells in vitro and xenograft tumors in vivo, leading to impaired function of CD8+ T cells. Furthermore, CXCR1/2 receptor depletion, CXCR2 receptor antagonist AZD5069 and IL-8 neutralizing antibody application also significantly reversed GCMSCs mediated immunosuppression, restoring the antitumor capacity of PD-1 antibody.

Conclusions

Our findings reveal that blocking GCMSCs-derived IL-8/CXCR2 pathway decreasing PD-L1 expression and lactate production, improving antitumor efficacy of anti-PD-1 immunotherapy, may be of value for the treatment of advanced gastric carcinoma.

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Data availability

The data supporting the findings of this study are available from the corresponding author upon reasonable request, e-mail: zhuwei@ujs.edu.cn.

Abbreviations

TME:

Tumor microenvironment

PD-1:

Programmed death-1

PD-L1:

Programmed death ligand-1

HK2:

Hexokinase II

PBMCs:

Peripheral blood mononuclear cells

MSCs:

Mesenchymal stem cells

GCMSCs:

Gastric cancer mesenchymal stem cells

BMMSCs:

Bone marrow mesenchymal stem cells

HIF-1α:

Hypoxia-inducible factor-1α

CXCR1:

C-X-C motif chemokine receptor 1

CXCR2:

C-X-C motif chemokine receptor 2

IL-8:

Interleukin-8

IL-1:

Interleukin-1

IFN-γ:

Interferon γ

References

  1. Janjigian YY, Bendell J, Calvo E, Kim JW, Ascierto PA, Sharma P, et al. CheckMate-032 study: efficacy and safety of nivolumab and nivolumab plus ipilimumab in patients with metastatic esophagogastric cancer. J Clin Oncol. 2018;36(28):2836–44. https://doi.org/10.1200/JCO.2017.76.6212.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Shitara K, Ozguroglu M, Bang YJ, Di Bartolomeo M, Mandala M, Ryu MH, et al. Pembrolizumab versus paclitaxel for previously treated, advanced gastric or gastro-oesophageal junction cancer (KEYNOTE-061): a randomised, open-label, controlled, phase 3 trial. Lancet. 2018;392(10142):123–33. https://doi.org/10.1016/S0140-6736(18)31257-1.

    Article  CAS  PubMed  Google Scholar 

  3. Nanda R, Chow LQ, Dees EC, Berger R, Gupta S, Geva R, et al. Pembrolizumab in patients with advanced triple-negative breast cancer: phase Ib KEYNOTE-012 study. J Clin Oncol. 2016;34(21):2460–7. https://doi.org/10.1200/JCO.2015.64.8931.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Wieser V, Adolph TE, Enrich B, Kuliopulos A, Kaser A, Tilg H, et al. Reversal of murine alcoholic steatohepatitis by pepducin-based functional blockade of interleukin-8 receptors. Gut. 2017;66(5):930–8. https://doi.org/10.1136/gutjnl-2015-310344.

    Article  CAS  PubMed  Google Scholar 

  5. Eruslanov EB, Bhojnagarwala PS, Quatromoni JG, Stephen TL, Ranganathan A, Deshpande C, et al. Tumor-associated neutrophils stimulate T cell responses in early-stage human lung cancer. J Clin Investig. 2014;124(12):5466–80. https://doi.org/10.1172/JCI77053.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Liu Y, Zhang Y, Wang S, Dong QZ, Shen Z, Wang W, et al. Prospero-related homeobox 1 drives angiogenesis of hepatocellular carcinoma through selectively activating interleukin-8 expression. Hepatology. 2017;66(6):1894–909. https://doi.org/10.1002/hep.29337.

    Article  CAS  PubMed  Google Scholar 

  7. Shang A, Gu C, Zhou C, Yang Y, Chen C, Zeng B, et al. Exosomal KRAS mutation promotes the formation of tumor-associated neutrophil extracellular traps and causes deterioration of colorectal cancer by inducing IL-8 expression. Cell Commun Signal. 2020;18(1):52. https://doi.org/10.1186/s12964-020-0517-1.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Siu MKY, Jiang YX, Wang JJ, Leung THY, Ngu SF, Cheung ANY, et al. PDK1 promotes ovarian cancer metastasis by modulating tumor-mesothelial adhesion, invasion, and angiogenesis via alpha5beta1 integrin and JNK/IL-8 signaling. Oncogenesis. 2020;9(2):24. https://doi.org/10.1038/s41389-020-0209-0.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Schalper KA, Carleton M, Zhou M, Chen T, Feng Y, Huang SP, et al. Elevated serum interleukin-8 is associated with enhanced intratumor neutrophils and reduced clinical benefit of immune-checkpoint inhibitors. Nat Med. 2020;26(5):688–92. https://doi.org/10.1038/s41591-020-0856-x.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Yuen KC, Liu LF, Gupta V, Madireddi S, Keerthivasan S, Li C, et al. High systemic and tumor-associated IL-8 correlates with reduced clinical benefit of PD-L1 blockade. Nat Med. 2020;26(5):693–8. https://doi.org/10.1038/s41591-020-0860-1.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Sun L, Wang Q, Chen B, Zhao Y, Shen B, Wang H, et al. Gastric cancer mesenchymal stem cells derived IL-8 induces PD-L1 expression in gastric cancer cells via STAT3/mTOR-c-Myc signal axis. Cell Death Dis. 2018;9(9):928. https://doi.org/10.1038/s41419-018-0988-9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Wang Q, Huang C, Ding Y, Wen S, Wang X, Guo S, et al. Inhibition of CCCTC Binding Factor-Programmed Cell Death Ligand 1 Axis Suppresses Emergence of Chemoresistance Induced by Gastric Cancer-Derived Mesenchymal Stem Cells. Front Immunol. 2022;13:884373. https://doi.org/10.3389/fimmu.2022.884373.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Donnelly JM, Engevik AC, Engevik M, Schumacher MA, Xiao C, Yang L, et al. Gastritis promotes an activated bone marrow-derived mesenchymal stem cell with a phenotype reminiscent of a cancer-promoting cell. Dig Dis Sci. 2014;59(3):569–82. https://doi.org/10.1007/s10620-013-2927-z.

    Article  PubMed  Google Scholar 

  14. Sun L, Wang Q, Chen B, Zhao Y, Shen B, Wang X, et al. Human gastric cancer mesenchymal stem cell-derived IL15 contributes to tumor cell epithelial-mesenchymal transition via upregulation tregs ratio and PD-1 expression in CD4(+)T cell. Stem Cells Dev. 2018;27(17):1203–14. https://doi.org/10.1089/scd.2018.0043.

    Article  CAS  PubMed  Google Scholar 

  15. Chen B, Cai T, Huang C, Zang X, Sun L, Guo S, et al. G6PD-NF-kappaB-HGF signal in gastric cancer-associated mesenchymal stem cells promotes the proliferation and metastasis of gastric cancer cells by upregulating the expression of HK2. Front Oncol. 2021;11:648706. https://doi.org/10.3389/fonc.2021.648706.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Shitara K, Ajani JA, Moehler M, Garrido M, Gallardo C, Shen L, et al. Nivolumab plus chemotherapy or ipilimumab in gastro-oesophageal cancer. Nature. 2022;603(7903):942–8. https://doi.org/10.1038/s41586-022-04508-4.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Gordan JD, Thompson CB, Simon MC. HIF and c-Myc: sibling rivals for control of cancer cell metabolism and proliferation. Cancer Cell. 2007;12(2):108–13. https://doi.org/10.1016/j.ccr.2007.07.006.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Wang ZH, Peng WB, Zhang P, Yang XP, Zhou Q. Lactate in the tumour microenvironment: From immune modulation to therapy. EBioMedicine. 2021;73:103627. https://doi.org/10.1016/j.ebiom.2021.103627.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Thomas GE, Egan G, Garcia-Prat L, Botham A, Voisin V, Patel PS, et al. The metabolic enzyme hexokinase 2 localizes to the nucleus in AML and normal haematopoietic stem and progenitor cells to maintain stemness. Nat Cell Biol. 2022;24(6):872–84. https://doi.org/10.1038/s41556-022-00925-9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Palsson-McDermott EM, Curtis AM, Goel G, Lauterbach MA, Sheedy FJ, Gleeson LE, et al. Pyruvate kinase M2 regulates Hif-1alpha activity and IL-1beta induction and is a critical determinant of the warburg effect in LPS-activated macrophages. Cell Metab. 2015;21(1):65–80. https://doi.org/10.1016/j.cmet.2014.12.005.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Ippolito L, Morandi A, Giannoni E, Chiarugi P. Lactate: a metabolic driver in the tumour landscape. Trends Biochem Sci. 2019;44(2):153–66. https://doi.org/10.1016/j.tibs.2018.10.011.

    Article  CAS  PubMed  Google Scholar 

  22. Watson MJ, Vignali PDA, Mullett SJ, Overacre-Delgoffe AE, Peralta RM, Grebinoski S, et al. Metabolic support of tumour-infiltrating regulatory T cells by lactic acid. Nature. 2021;591(7851):645–51. https://doi.org/10.1038/s41586-020-03045-2.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Xin Q, Wang H, Li Q, Liu S, Qu K, Liu C, et al. Lactylation: a passing fad or the future of posttranslational modification. Inflammation. 2022;45(4):1419–29. https://doi.org/10.1007/s10753-022-01637-w.

    Article  CAS  PubMed  Google Scholar 

  24. Liu H, Zhao Q, Tan L, Wu X, Huang R, Zuo Y, et al. Neutralizing IL-8 potentiates immune checkpoint blockade efficacy for glioma. Cancer Cell. 2023;41(4):693-710 e8. https://doi.org/10.1016/j.ccell.2023.03.004.

    Article  CAS  PubMed  Google Scholar 

  25. Wang M, Chen B, Sun XX, Zhao XD, Zhao YY, Sun L, et al. Gastric cancer tissue-derived mesenchymal stem cells impact peripheral blood mononuclear cells via disruption of Treg/Th17 balance to promote gastric cancer progression. Exp Cell Res. 2017;361(1):19–29. https://doi.org/10.1016/j.yexcr.2017.09.036.

    Article  CAS  PubMed  Google Scholar 

  26. Wang M, Zhao X, Qiu R, Gong Z, Huang F, Yu W, et al. Lymph node metastasis-derived gastric cancer cells educate bone marrow-derived mesenchymal stem cells via YAP signaling activation by exosomal Wnt5a. Oncogene. 2021;40(12):2296–308. https://doi.org/10.1038/s41388-021-01722-8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Gao QZ, Cui LJ, Huang C, Zhou CL, Chen B, Wang QQ, et al. Gastric cancer-derived exosomes induce PD-L1 expression on human bone marrow mesenchymal stem cells through the AKT-c-Myc signal axis. All Life. 2022;15(1):442–51. https://doi.org/10.1080/26895293.2022.2058098.

    Article  CAS  Google Scholar 

  28. Sun L, Huang C, Zhu M, Guo S, Gao Q, Wang Q, et al. Gastric cancer mesenchymal stem cells regulate PD-L1–CTCF enhancing cancer stem cell-like properties and tumorigenesis. Theranostics. 2020;10(26):11950–62. https://doi.org/10.7150/thno.49717.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Zhai J, Shen J, Xie G, Wu J, He M, Gao L, et al. Cancer-associated fibroblasts-derived IL-8 mediates resistance to cisplatin in human gastric cancer. Cancer Lett. 2019;454:37–43. https://doi.org/10.1016/j.canlet.2019.04.002.

    Article  CAS  PubMed  Google Scholar 

  30. Wu TD, Madireddi S, de Almeida PE, Banchereau R, Chen YJ, Chitre AS, et al. Peripheral T cell expansion predicts tumour infiltration and clinical response. Nature. 2020;579(7798):274–8. https://doi.org/10.1038/s41586-020-2056-8.

    Article  CAS  PubMed  Google Scholar 

  31. Lee YS, Choi I, Ning Y, Kim NY, Khatchadourian V, Yang D, et al. Interleukin-8 and its receptor CXCR2 in the tumour microenvironment promote colon cancer growth, progression and metastasis. Br J Cancer. 2012;106(11):1833–41. https://doi.org/10.1038/bjc.2012.177.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Jeffery CJ. Moonlighting proteins. Trends Biochem Sci. 1999;24(1):8–11. https://doi.org/10.1016/s0968-0004(98)01335-8.

    Article  CAS  PubMed  Google Scholar 

  33. Wang Z, Li M, Jiang H, Luo S, Shao F, Xia Y, et al. Fructose-1,6-bisphosphatase 1 functions as a protein phosphatase to dephosphorylate histone H3 and suppresses PPARalpha-regulated gene transcription and tumour growth. Nat Cell Biol. 2022;24(11):1655–65. https://doi.org/10.1038/s41556-022-01009-4.

    Article  CAS  PubMed  Google Scholar 

  34. Yang W, Xia Y, Hawke D, Li X, Liang J, Xing D, et al. PKM2 phosphorylates histone H3 and promotes gene transcription and tumorigenesis. Cell. 2012;150(4):685–96. https://doi.org/10.1016/j.cell.2012.07.018.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Roberts DJ, Tan-Sah VP, Smith JM, Miyamoto S. Akt phosphorylates HK-II at Thr-473 and increases mitochondrial HK-II association to protect cardiomyocytes. J Biol Chem. 2013;288(33):23798–806. https://doi.org/10.1074/jbc.M113.482026.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Li H, Lu S, Chen Y, Zheng L, Chen L, Ding H, et al. AKT2 phosphorylation of hexokinase 2 at T473 promotes tumorigenesis and metastasis in colon cancer cells via NF-kappaB, HIF1alpha, MMP2, and MMP9 upregulation. Cell Signal. 2019;58:99–110. https://doi.org/10.1016/j.cellsig.2019.03.011.

    Article  CAS  PubMed  Google Scholar 

  37. Chen G, Huang AC, Zhang W, Zhang G, Wu M, Xu W, et al. Exosomal PD-L1 contributes to immunosuppression and is associated with anti-PD-1 response. Nature. 2018;560(7718):382–6. https://doi.org/10.1038/s41586-018-0392-8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Le DT, Durham JN, Smith KN, Wang H, Bartlett BR, Aulakh LK, et al. Mismatch repair deficiency predicts response of solid tumors to PD-1 blockade. Science. 2017;357(6349):409–13. https://doi.org/10.1126/science.aan6733.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

The authors thank AstraZeneca (China) Co. Ltd., for providing AZD5069 drug during the research.

Funding

This work was supported by the National Natural Science Foundation of China No. 81972313 (WZ), No. 81972822 (BS), No. 82203547 (LS), the Bethune Charitable Foundation No. G-X-2019-0101-12 (BS).

Author information

Author notes

  1. Chao Huang and Bin Chen contributed equally to this work.

  2. Bo Shen and Wei Zhu jointly supervised this work.

Authors and Affiliations

  1. School of Medicine, Jiangsu University, 301 Xuefu Road, Jingkou District, Zhenjiang, Jiangsu, China

    Chao Huang, Bin Chen, Li Sun, Yuanyuan Zhao, Qiuzhi Gao, Qianqian Wang, Mei Wang, Xu Zhang, Wenrong Xu & Wei Zhu

  2. Department of Oncology, Jiangsu Cancer Hospital and Jiangsu Institute of Cancer Research and, The Affiliated Cancer Hospital of Nanjing Medical University, Nanjing, Jiangsu, China

    Xin Wang & Bo Shen

  3. Department of Laboratory Medicine, Affiliated Taizhou People’s Hospital of Nanjing Medical University, Taizhou, Jiangsu, China

    Juan Xu & Chenglin Zhou

  4. Department of Oncology, Affiliated Hospital of Jiangsu University, Zhenjiang, Jiangsu, China

    Deqiang Wang

  5. Department of Gastrointestinal Surgery, Affiliated People’s Hospital of Jiangsu University, Zhenjiang, Jiangsu, China

    Zhihong Chen

  6. Department of Clinical Laboratory, Affiliated Hospital of Jining Medical University, Jining, Shandong, China

    Bin Chen

  7. Department of Clinical Laboratory, Kunshan First People′s Hospital, Kunshan, China

    Li Sun

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  1. Chao Huang
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  3. Xin Wang
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Contributions

CH and BC: conceptualization, methodology, validation, data curation, formal analysis, writing—original draft preparation. YYZ, QQW, LS, and QZG: validation, formal analysis. ZHC, CLZ, DQW, JX, XW: resources. MW, XZ and WRX: conceptualization. WZ and BS: writing—review and editing, supervision, funding acquisition.

Corresponding author

Correspondence to Wei Zhu.

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Conflict of interest

The authors declare no potential conflicts of interest.

Ethical approval

This study involves human subjects, and all samples were obtained following individual informed consent and ethical approval by the Ethics Committee of the Affiliated Cancer Hospital of Nanjing Medical University ID#: 2021ke-010. The subjects gave informed consent to participate in the study before taking part. The animal study was approved by the Ethics Committee of Jiangsu University. All procedures were conducted following the guidelines of the National Institute of Health regarding the care and use of laboratory animals (NIH Publication No. 8023, revised 1978) ID#: UJS-IACUC-AP-2022022812.

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Huang, C., Chen, B., Wang, X. et al. Gastric cancer mesenchymal stem cells via the CXCR2/HK2/PD-L1 pathway mediate immunosuppression. Gastric Cancer 26, 691–707 (2023). https://doi.org/10.1007/s10120-023-01405-1

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