Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Cell fusion upregulates PD-L1 expression for evasion from immunosurveillance

Cancer Gene Therapy volume 31, pages 158–173 (2024)Cite this article

Subjects

Abstract

MSCs (mesenchymal stem cells), responsible for tissue repair, rarely undergo cell fusion with somatic cells. Here, we show that ~5% of bladder cancer cells (UMUC-3) fuses with bone marrow-derived MSC (BM-MSC) in co-culture and maintains high tumorigenicity. In eleven fusion cell clones that have been established, Mb-scale deletions carried by the bladder cancer cells are mostly absent in the fusion cells, but copy number gains contributed by the cancer cells have stayed. Fusion cells exhibit increased populations of mitotic cells with 3-polar spindles, indicative of genomic instability. They grow faster in vitro and exhibit higher colony formation in anchorage-independent growth assay in soft agar than the parent UMUC-3 does. Fusion cells develop tumors, after 4 weeks of time lag, as efficiently as the parent UMUC-3 does in xenograft experiments. 264 genes are identified whose expression is specifically altered in the fusion cells. Many of them are interferon-stimulated genes (ISG), but are activated in a manner independent of interferon. Among them, we show that PD-L1 is induced in fusion cells, and its knockout decreases tumorigenesis in a xenograft model. PD-L1 is induced in a manner independent of STAT1 known to regulate PD-L1 expression, but is regulated by histone modification, and is likely to inhibit phagocytosis by PD1-expressing macrophages, thus protecting cancer cells from immunological attacks. The fusion cells overexpress multiple cytokines including CCL2 that cause tumor progression by converting infiltrating macrophages to tumor-associated-macrophage (TAM). The results present mechanisms of how cell fusion promotes tumorigenesis, revealing a novel link between cell fusion and PD-L1, and underscore the efficacy of cancer immunotherapy.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: UMUC-3 cells (human urothelial carcinoma) fuse with BM-MSC to form fusion cells with full chromosome complements.
Fig. 2: Genome stability of fusion cells.
Fig. 3: Fusion cells are tumorigenic in vitro and in vivo.
Fig. 4: Genes involved in type I interferon signal pathway are activated in fusion cells.
Fig. 5: Cell fusion-induced upregulation of PD-L1 expression.
Fig. 6: PD-L1 expression in fusion cells is reduced by inhibition of BRD4.
Fig. 7: PD-L1 knockdown in the fusion cells results in decreased tumorigenicity in xenograft model.

Similar content being viewed by others

Data availability

The data that support the finding of this study are openly available in figshare at https://doi.org/10.6084/m9.figshare.20140976 and in bioRxiv at https://doi.org/10.1101/2022.06.14.496068.

References

  1. Hernández JM, Podbilewicz B. The hallmarks of cell-cell fusion. Development. 2017;144:4481–95.

    Article  PubMed  Google Scholar 

  2. Zhou X, Merchak K, Lee W, Grande JP, Cascalho M, Platt JL. Cell fusion connects oncogenesis with tumor evolution. Am J Pathol. 2015;185:2049–60.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  3. Oren-Suissa M, Podbilewicz B. Cell fusion during development. Trends Cell Biol. 2007;17:537–46.

    Article  PubMed  CAS  Google Scholar 

  4. Weiler J, Dittmar T. Cell fusion in human cancer: the dark matter hypothesis. Cells. 2019;8:132. https://doi.org/10.3390/cells8020132.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  5. Gast CE, Silk AD, Zarour L, Riegler L, Burkhart JG, Gustafson KT, et al. Cell fusion potentiates tumor heterogeneity and reveals circulating hybrid cells that correlate with stage and survival. Sci Adv. 2018;4:eaat7828.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Yin L, Hu P, Shi X, Qian W, Zhau HE, Pandol SJ, et al. Cancer cell’s neuroendocrine feature can be acquired through cell-cell fusion during cancer-neural stem cell interaction. Sci Rep. 2020;10:1216.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  7. Zhang LN, Zhang DD, Yang L, Gu YX, Zuo QP, Wang HY, et al. Roles of cell fusion between mesenchymal stromal/stem cells and malignant cells in tumor growth and metastasis. FEBS J. 2021;288:1447–56.

    Article  PubMed  CAS  Google Scholar 

  8. Pittenger MF, Discher DE, Péault BM, Phinney DG, Hare JM, Caplan AI. Mesenchymal stem cell perspective: cell biology to clinical progress. npj Regenerative Med. 2019;4:22.

    Article  Google Scholar 

  9. Karnoub AE, Dash AB, Vo AP, Sullivan A, Brooks MW, Bell GW, et al. Mesenchymal stem cells within tumour stroma promote breast cancer metastasis. Nature. 2007;449:557–63.

    Article  PubMed  CAS  Google Scholar 

  10. Alvarez-Dolado M, Pardal R, Garcia-Verdugo JM, Fike JR, Lee HO, Pfeffer K, et al. Fusion of bone-marrow-derived cells with Purkinje neurons, cardiomyocytes and hepatocytes. Nature. 2003;425:968–73.

    Article  PubMed  CAS  Google Scholar 

  11. Nygren JM, Jovinge S, Breitbach M, Säwén P, Röll W, Hescheler J, et al. Bone marrow-derived hematopoietic cells generate cardiomyocytes at a low frequency through cell fusion, but not transdifferentiation. Nat Med. 2004;10:494–501.

    Article  PubMed  CAS  Google Scholar 

  12. Rizvi AZ, Swain JR, Davies PS, Bailey AS, Decker AD, Willenbring H, et al. Bone marrow-derived cells fuse with normal and transformed intestinal stem cells. Proc Natl Acad Sci USA. 2006;103:6321–5.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  13. Pawelek JM, Chakraborty AK. Fusion of tumour cells with bone marrow-derived cells: a unifying explanation for metastasis. Nat Rev Cancer. 2008;8:377–86.

    Article  PubMed  CAS  Google Scholar 

  14. Johansson CB, Youssef S, Koleckar K, Holbrook C, Doyonnas R, Corbel SY, et al. Extensive fusion of haematopoietic cells with Purkinje neurons in response to chronic inflammation. Nat Cell Biol. 2008;10:575–83.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  15. Rappa G, Mercapide J, Lorico A. Spontaneous formation of tumorigenic hybrids between breast cancer and multipotent stromal cells is a source of tumor heterogeneity. Am J Pathol. 2012;180:2504–15.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  16. Nygren JM, Liuba K, Breitbach M, Stott S, Thorén L, Roell W, et al. Myeloid and lymphoid contribution to non-haematopoietic lineages through irradiation-induced heterotypic cell fusion. Nat Cell Biol. 2008;10:584–92.

    Article  PubMed  CAS  Google Scholar 

  17. Delespaul L, Merle C, Lesluyes T, Lagarde P, Le Guellec S, Perot G, et al. Fusion-mediated chromosomal instability promotes aneuploidy patterns that resemble human tumors. Oncogene. 2019;38:6083–94.

    Article  PubMed  CAS  Google Scholar 

  18. Feliciano D, Ott CM, Espinosa-Medina I, Weigel AV, Benedetti L, Milano KM, et al. YAP1 nuclear efflux and transcriptional reprograming follow membrane diminution upon VSV-G-induced cell fusion. Nat Commun. 2021;12:4502.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  19. Ku JWK, Chen Y, Lim BJW, Gasser S, Crasta KC, Gan YH. Bacterial-induced cell fusion is a danger signal triggering cGAS-STING pathway via micronuclei formation. Proc Natl Acad Sci USA. 2020;117:15923–34.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  20. Holm CK, Jensen SB, Jakobsen MR, Cheshenko N, Horan KA, Moeller HB, et al. Virus-cell fusion as a trigger of innate immunity dependent on the adaptor STING. Nat Immunol. 2012;13:737–43.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  21. Schoggins JW. Interferon-Stimulated Genes: What Do They All Do? Annu Rev Virol. 2019;6:567–84.

    Article  PubMed  CAS  Google Scholar 

  22. Kim D, Pertea G, Trapnell C, Pimentel H, Kelley R, Salzberg SL. TopHat2: accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions. Genome Biol. 2013;14:R36.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 2014;15:550.

    Article  PubMed  PubMed Central  Google Scholar 

  24. Benjamini Y, Hochberg Y. Controlling the false discovery rate: a practical and powerful approach to multiple testing. J R Stat Soc Series B. (Methodological). 1995;57:289–300.

  25. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, et al. Multilineage potential of adult human mesenchymal stem cells. Science. 1999;284:143–7.

    Article  PubMed  CAS  Google Scholar 

  26. Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini FC, Krause DS, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006;8:315–7.

    Article  PubMed  CAS  Google Scholar 

  27. Tajima Y, Saito S, Ohno K, Tsukimura T, Tsujino S, Sakuraba H. Biochemical and structural study on a S529V mutant acid alpha-glucosidase responsive to pharmacological chaperones. J Hum Genet. 2011;56:440–6.

    Article  PubMed  CAS  Google Scholar 

  28. Sanjana NE, Shalem O, Zhang F. Improved vectors and genome-wide libraries for CRISPR screening. Nat Methods. 2014;11:783–4.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  29. Tomayko MM, Reynolds CP. Determination of subcutaneous tumor size in athymic (nude) mice. Cancer Chemother Pharmacol. 1989;24:148–54.

    Article  PubMed  CAS  Google Scholar 

  30. Baker MJ, Goldstein AM, Gordon PL, Harbaugh KS, Mackley HB, Glantz MJ, et al. An interstitial deletion within 9p21.3 and extending beyond CDKN2A predisposes to melanoma, neural system tumours and possible haematological malignancies. J Med Genet. 2016;53:721–7.

    Article  PubMed  CAS  Google Scholar 

  31. Frigerio S, Disciglio V, Manoukian S, Peissel B, Della Torre G, Maurichi A, et al. A large de novo9p21.3 deletion in a girl affected by astrocytoma and multiple melanoma. BMC Med Genet. 2014;15:59.

    Article  PubMed  PubMed Central  Google Scholar 

  32. Tanaka M, Grossman HB. In vivo gene therapy of human bladder cancer with PTEN suppresses tumor growth, downregulates phosphorylated Akt, and increases sensitivity to doxorubicin. Gene Ther. 2003;10:1636–42.

    Article  PubMed  CAS  Google Scholar 

  33. Aguirre-Ghiso JA, Sosa MS. Emerging topics on disseminated cancer cell dormancy and the paradigm of metastasis. Annu Rev Cancer Biol. 2018;2:377–93.

    Article  Google Scholar 

  34. Li X, Yao W, Yuan Y, Chen P, Li B, Li J, et al. Targeting of tumour-infiltrating macrophages via CCL2/CCR2 signalling as a therapeutic strategy against hepatocellular carcinoma. Gut. 2017;66:157–67.

    Article  PubMed  CAS  Google Scholar 

  35. BÜHring H-J, Battula VL, Treml S, Schewe B, Kanz L, Vogel W. Novel markers for the prospective isolation of human MSC. Ann N Y Acad Sci. 2007;1106:262–71. https://doi.org/10.1196/annals.1392.000

    Article  PubMed  CAS  Google Scholar 

  36. Miwa H, Era T. Tracing the destiny of mesenchymal stem cells from embryo to adult bone marrow and white adipose tissue via Pdgfrα expression. Development. 2018;145:dev155879. https://doi.org/10.1242/dev.155879.

    Article  PubMed  CAS  Google Scholar 

  37. Soliman H, Theret M, Scott W, Hill L, Underhill TM, Hinz B, et al. Multipotent stromal cells: One name, multiple identities. Cell Stem Cell. 2021;28:1690–707.

    Article  PubMed  CAS  Google Scholar 

  38. Miyamoto H, Yang Z, Chen Y-T, Ishiguro H, Uemura H, Kubota Y, et al. Promotion of bladder cancer development and progression by androgen receptor signals. JNCI: J Natl Cancer Inst. 2007;99:558–68.

    Article  PubMed  CAS  Google Scholar 

  39. Burr ML, Sparbier CE, Chan Y-C, Williamson JC, Woods K, Beavis PA, et al. CMTM6 maintains the expression of PD-L1 and regulates anti-tumour immunity. Nature. 2017;549:101–5.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  40. Mezzadra R, Sun C, Jae LT, Gomez-Eerland R, de Vries E, Wu W, et al. Identification of CMTM6 and CMTM4 as PD-L1 protein regulators. Nature. 2017;549:106–10.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  41. Zhang J, Bu X, Wang H, Zhu Y, Geng Y, Nihira NT, et al. Cyclin D–CDK4 kinase destabilizes PD-L1 via cullin 3–SPOP to control cancer immune surveillance. Nature. 2018;553:91–95.

    Article  PubMed  CAS  Google Scholar 

  42. Li C-W, Lim S-O, Xia W, Lee H-H, Chan L-C, Kuo C-W, et al. Glycosylation and stabilization of programmed death ligand-1 suppresses T-cell activity. Nat Commun. 2016;7:12632.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  43. Garcia-Diaz A, Shin DS, Moreno BH, Saco J, Escuin-Ordinas H, Rodriguez GA, et al. Interferon receptor signaling pathways regulating PD-L1 and PD-L2 expression. Cell Rep. 2019;29:3766.

    Article  PubMed  CAS  Google Scholar 

  44. Kwak G, Kim D, Nam GH, Wang SY, Kim IS, Kim SH, et al. Programmed cell death protein ligand-1 silencing with polyethylenimine-dermatan sulfate complex for dual inhibition of melanoma growth. ACS Nano. 2017;11:10135–46.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  45. Pelleitier M, Montplaisir S. The nude mouse: a model of deficient T-cell function. Methods Achiev Exp Pathol. 1975;7:149–66.

    PubMed  CAS  Google Scholar 

  46. Gordon SR, Maute RL, Dulken BW, Hutter G, George BM, McCracken MN, et al. PD-1 expression by tumour-associated macrophages inhibits phagocytosis and tumour immunity. Nature. 2017;545:495–9.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  47. Qian BZ, Li J, Zhang H, Kitamura T, Zhang J, Campion LR, et al. CCL2 recruits inflammatory monocytes to facilitate breast-tumour metastasis. Nature. 2011;475:222–5.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  48. He G, Dhar D, Nakagawa H, Font-Burgada J, Ogata H, Jiang Y, et al. Identification of liver cancer progenitors whose malignant progression depends on autocrine IL-6 signaling. Cell. 2013;155:384–96.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  49. Sun C, Mezzadra R, Schumacher TN. Regulation and Function of the PD-L1 Checkpoint. Immunity. 2018;48:434–52.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  50. Li CW, Lim SO, Chung EM, Kim YS, Park AH, Yao J, et al. Eradication of triple-negative breast cancer cells by targeting glycosylated PD-L1. Cancer Cell. 2018;33:187–201.e110.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  51. Zhang J, Bu X, Wang H, Zhu Y, Geng Y, Nihira NT, et al. Cyclin D-CDK4 kinase destabilizes PD-L1 via cullin 3-SPOP to control cancer immune surveillance. Nature. 2018;553:91–95.

    Article  PubMed  CAS  Google Scholar 

  52. Lim SO, Li CW, Xia W, Cha JH, Chan LC, Wu Y, et al. Deubiquitination and Stabilization of PD-L1 by CSN5. Cancer Cell. 2016;30:925–39.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  53. Chan LC, Li CW, Xia W, Hsu JM, Lee HH, Cha JH, et al. IL-6/JAK1 pathway drives PD-L1 Y112 phosphorylation to promote cancer immune evasion. J Clin Investig. 2019;129:3324–38.

    Article  PubMed  PubMed Central  Google Scholar 

  54. Sun LL, Yang RY, Li CW, Chen MK, Shao B, Hsu JM, et al. Inhibition of ATR downregulates PD-L1 and sensitizes tumor cells to T cell-mediated killing. Am J Cancer Res. 2018;8:1307–16.

    PubMed  PubMed Central  CAS  Google Scholar 

  55. Maniecki MB, Etzerodt A, Ulhøi BP, Steiniche T, Borre M, Dyrskjøt L, et al. Tumor-promoting macrophages induce the expression of the macrophage-specific receptor CD163 in malignant cells. Int J Cancer. 2012;131:2320–31.

    Article  PubMed  CAS  Google Scholar 

  56. Rubio C, Avendaño-Ortiz J, Ruiz-Palomares R, Karaivanova V, Alberquilla O, Sánchez-Domínguez R, et al. Toward tumor fight and tumor microenvironment remodeling: PBA induces cell cycle arrest and reduces tumor hybrid cells’ pluripotency in bladder cancer. Cancers. 2022;14:287.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  57. Yu H, Liu T, Zhao Z, Chen Y, Zeng J, Liu S, et al. Mutations in 3′-long terminal repeat of HERV-W family in chromosome 7 upregulate syncytin-1 expression in urothelial cell carcinoma of the bladder through interacting with c-Myb. Oncogene. 2014;33:3947–58.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

We thank Yoshinobu Iguchi and Kazunari Sekiyama for the preparation of cryosections. We also thank Yasumasa Nishito for assistance in the analyses with array-CGH, Daisuke Yamane for insightful discussion and comments. We thank Hiroyuki Sasanuma for critical reading of the manuscript. We thank the members of our laboratory for helpful discussion. This work was supported by JSPS KAKENHI Grant-in-Aid for Scientific Research (A) Grant Number 20H00463 (to HM) and by KAKENHI Grant-in-Aid for Scientific Research (C) Grant Number 18K07220 (to YT) and KAKENHI Grant-in-Aid for Challenging Exploratory Research Grant Number 16K15262 (to YT), respectively.

Author information

Authors and Affiliations

  1. Genome Dynamics Project, Department of Basic Medical Sciences, Tokyo Metropolitan Institute of Medical Science, Kamikitazawa, Setagaya-ku, Tokyo, 156-8506, Japan

    Youichi Tajima & Hisao Masai

  2. Center for Medical Research Cooperation, Tokyo Metropolitan Institute of Medical Science, Kamikitazawa, Setagaya-ku, Tokyo, 156-8506, Japan

    Futoshi Shibasaki

Authors
  1. Youichi Tajima
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Futoshi Shibasaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hisao Masai
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Conceptualization: YT and HM; Methodology and Investigation: YT; Writing and Visualization: YT and HM; Supervision: HM and FS; Funding Acquisition and Project Administration: HM.

Corresponding authors

Correspondence to Youichi Tajima or Hisao Masai.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Tajima, Y., Shibasaki, F. & Masai, H. Cell fusion upregulates PD-L1 expression for evasion from immunosurveillance. Cancer Gene Ther 31, 158–173 (2024). https://doi.org/10.1038/s41417-023-00693-0

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41417-023-00693-0

Search

Advanced search

Quick links