Sublethal Radiation Affects Antigen Processing and Presentation Genes to Enhance Immunogenicity of Cancer Cells
Achamaporn Punnanitinont, Eric D Kannisto, Junko Matsuzaki, Kunle Odunsi, Sai Yendamuri, Anurag K Singh, Santosh K Patnaik, Achamaporn Punnanitinont, Eric D Kannisto, Junko Matsuzaki, Kunle Odunsi, Sai Yendamuri, Anurag K Singh, Santosh K Patnaik
Abstract
While immunotherapy in cancer is designed to stimulate effector T cell response, tumor-associated antigens have to be presented on malignant cells at a sufficient level for recognition of cancer by T cells. Recent studies suggest that radiotherapy enhances the anti-cancer immune response and also improves the efficacy of immunotherapy. To understand the molecular basis of such observations, we examined the effect of ionizing X-rays on tumor antigens and their presentation in a set of nine human cell lines representing cancers of the esophagus, lung, and head and neck. A single dose of 7.5 or 15 Gy radiation enhanced the New York esophageal squamous cell carcinoma 1 (NY-ESO-1) tumor-antigen-mediated recognition of cancer cells by NY-ESO-1-specific CD8+ T cells. Irradiation led to significant enlargement of live cells after four days, and microscopy and flow cytometry revealed multinucleation and polyploidy in the cells because of dysregulated mitosis, which was also revealed in RNA-sequencing-based transcriptome profiles of cells. Transcriptome analyses also showed that while radiation had no universal effect on genes encoding tumor antigens, it upregulated the expression of numerous genes involved in antigen processing and presentation pathways in all cell lines. This effect may explain the immunostimulatory role of cancer radiotherapy.
Keywords: antigen presentation; cancer cell line; gene expression; head and neck cancer; lung cancer; radiation; tumor antigen.
Conflict of interest statement
The authors declare no conflict of interest.
Figures
References
- Stone H.B., Peters L.J., Milas L. Effect of host immune capability on radiocurability and subsequent transplantability of a murine fibrosarcoma. J. Natl. Cancer Inst. 1979;63:1229–1235.
- Meng X., Feng R., Yang L., Xing L., Yu J. The role of radiation oncology in immuno-oncology. Oncologist. 2019;24:S42–S52. doi: 10.1634/theoncologist.2019-IO-S1-s04.
- Solanki A.A., Bossi A., Efstathiou J.A., Lock D., Mondini M., Ramapriyan R., Welsh J., Kang J. Combining immunotherapy with radiotherapy for the treatment of genitourinary malignancies. Eur. Urol. Oncol. 2019;2:79–87. doi: 10.1016/j.euo.2018.09.013.
- Zhou C., Zhang J. Immunotherapy-based combination strategies for treatment of gastrointestinal cancers: Current status and future prospects. Front. Med. 2019;13:12–23. doi: 10.1007/s11684-019-0685-9.
- Lee L., Matulonis U. Immunotherapy and radiation combinatorial trials in gynecologic cancer: A potential synergy? Gynecol. Oncol. 2019;154:236–245. doi: 10.1016/j.ygyno.2019.03.255.
- Ko E.C., Raben D., Formenti S.C. The integration of radiotherapy with immunotherapy for the treatment of non-small cell lung cancer. Clin. Cancer Res. 2018;24:5792–5806. doi: 10.1158/1078-0432.CCR-17-3620.
- Karam S.D., Raben D. Radioimmunotherapy for the treatment of head and neck cancer. Lancet Oncol. 2019;20:e404–e416. doi: 10.1016/S1470-2045(19)30306-7.
- Gameiro S.R., Jammeh M.L., Wattenberg M.M., Tsang K.Y., Ferrone S., Hodge J.W. Radiation-induced immunogenic modulation of tumor enhances antigen processing and calreticulin exposure, resulting in enhanced T-cell killing. Oncotarget. 2014;5:403–416. doi: 10.18632/oncotarget.1719.
- Kaur P., Asea A. Radiation-induced effects and the immune system in cancer. Front. Oncol. 2012;2:191. doi: 10.3389/fonc.2012.00191.
- Lhuillier C., Rudqvist N.P., Elemento O., Formenti S.C., Demaria S. Radiation therapy and anti-tumor immunity: Exposing immunogenic mutations to the immune system. Genome Med. 2019;11:40. doi: 10.1186/s13073-019-0653-7.
- Spiotto M., Fu Y.X., Weichselbaum R.R. The intersection of radiotherapy and immunotherapy: Mechanisms and clinical implications. Sci. Immunol. 2016;1 doi: 10.1126/sciimmunol.aag1266.
- Natale N., Reiner J., Southam C.M. Immunogenic efficacy of various syngeneic tumor cell preparations. Cancer. 1971;28:1118–1125. doi: 10.1002/1097-0142(1971)28:5<1118::AID-CNCR2820280505>;2-7.
- Wennerberg E., Vanpouille-Box C., Bornstein S., Yamazaki T., Demaria S., Galluzzi L. Immune recognition of irradiated cancer cells. Immunol. Rev. 2017;280:220–230. doi: 10.1111/imr.12568.
- Thomas R., Al-Khadairi G., Roelands J., Hendrickx W., Dermime S., Bedognetti D., Decock J. NY-ESO-1 based immunotherapy of cancer: Current perspectives. Front. Immunol. 2018;9:947. doi: 10.3389/fimmu.2018.00947.
- Matsuzaki J., Tsuji T., Luescher I.F., Shiku H., Mineno J., Okamoto S., Old L.J., Shrikant P., Gnjatic S., Odunsi K. Direct tumor recognition by a human CD4(+) T-cell subset potently mediates tumor growth inhibition and orchestrates anti-tumor immune responses. Sci. Rep. 2015;5:14896. doi: 10.1038/srep14896.
- Chueh A.C., Liew M.S., Russell P.A., Walkiewicz M., Jayachandran A., Starmans M.H.W., Boutros P.C., Wright G., Barnett S.A., Mariadason J.M., et al. Promoter hypomethylation of NY-ESO-1, association with clinicopathological features and PD-L1 expression in non-small cell lung cancer. Oncotarget. 2017;8:74036–74048. doi: 10.18632/oncotarget.18198.
- Singh A.K., Winslow T.B., Kermany M.H., Goritz V., Heit L., Miller A., Hoffend N.C., Stein L.C., Kumaraswamy L.K., Warren G.W., et al. A pilot study of stereotactic body radiation therapy combined with cytoreductive nephrectomy for metastatic renal cell carcinoma. Clin. Cancer Res. 2017;23:5055–5065. doi: 10.1158/1078-0432.CCR-16-2946.
- Eleftheriadou I., Brett S., Domogala A., Patasic L., Kijewska M.A., Soor K., Georgouli M., Dopierala J., Fisher P., Jing J., et al. 1229P—NY-ESO-1 and LAGE1A: An emerging target for cell therapies in solid tumours. Ann. Oncol. 2019;30:v503. doi: 10.1093/annonc/mdz253.055.
- Amundson S.A., Do K.T., Vinikoor L.C., Lee R.A., Koch-Paiz C.A., Ahn J., Reimers M., Chen Y., Scudiero D.A., Weinstein J.N., et al. Integrating global gene expression and radiation survival parameters across the 60 cell lines of the National Cancer Institute Anticancer Drug Screen. Cancer Res. 2008;68:415–424. doi: 10.1158/0008-5472.CAN-07-2120.
- Rangan S.R. A new human cell line (FaDu) from a hypopharyngeal carcinoma. Cancer. 1972;29:117–121. doi: 10.1002/1097-0142(197201)29:1<117::AID-CNCR2820290119>;2-R.
- Cowley G.S., Weir B.A., Vazquez F., Tamayo P., Scott J.A., Rusin S., East-Seletsky A., Ali L.D., Gerath W.F., Pantel S.E., et al. Parallel genome-scale loss of function screens in 216 cancer cell lines for the identification of context-specific genetic dependencies. Sci. Data. 2014;1:140035. doi: 10.1038/sdata.2014.35.
- Shao Y., Quan F., Li H.H., Yao X.B., Zhao Q., Zhao R.M. The radiosensitizing effect of resveratrol on hopypharyngeal carcinoma cell line FADU and its effect on the cell cycle. Zhongguo Zhong Xi Yi Jie He Za Zhi. 2015;35:699–703.
- Song S.Y., Das A.K., Minna J.D. Comparison between concurrent and sequential chemoradiation for non-small cell lung cancer in vitro. Oncol. Lett. 2014;7:307–310. doi: 10.3892/ol.2013.1707.
- Shintani S., Mihara M., Li C., Nakahara Y., Hino S., Nakashiro K., Hamakawa H. Up-regulation of DNA-dependent protein kinase correlates with radiation resistance in oral squamous cell carcinoma. Cancer Sci. 2003;94:894–900. doi: 10.1111/j.1349-7006.2003.tb01372.x.
- Zhang M., Rose B., Lee C.S., Hong A.M. In vitro 3-dimensional tumor model for radiosensitivity of HPV positive OSCC cell lines. Cancer Biol. Ther. 2015;16:1231–1240. doi: 10.1080/15384047.2015.1056410.
- Lu Y.C., Weng W.C., Lee H. Functional roles of calreticulin in cancer biology. Biomed. Res. Int. 2015;2015:526524. doi: 10.1155/2015/526524.
- Syrkina M.S., Rubtsov M.A. MUC1 in cancer immunotherapy—New hope or phantom menace? Biochemistry (Moscow) 2019;84:773–781. doi: 10.1134/S0006297919070083.
- Hanzelmann S., Castelo R., Guinney J. GSVA: Gene set variation analysis for microarray and RNA-seq data. BMC Bioinform. 2013;14:7. doi: 10.1186/1471-2105-14-7.
- Kelly A., Trowsdale J. Genetics of antigen processing and presentation. Immunogenetics. 2019;71:161–170. doi: 10.1007/s00251-018-1082-2.
- Wang J.-S., Wang H.-J., Qian H.-L. Biological effects of radiation on cancer cells. Mil. Med. Res. 2018;5:20. doi: 10.1186/s40779-018-0167-4.
- Barcellos-Hoff M.H., Park C., Wright E.G. Radiation and the microenvironment—Tumorigenesis and therapy. Nat. Rev. Cancer. 2005;5:867–875. doi: 10.1038/nrc1735.
- Frey B., Rubner Y., Kulzer L., Werthmoller N., Weiss E.M., Fietkau R., Gaipl U.S. Antitumor immune responses induced by ionizing irradiation and further immune stimulation. Cancer Immunol. Immunother. 2014;63:29–36. doi: 10.1007/s00262-013-1474-y.
- Chajon E., Castelli J., Marsiglia H., De Crevoisier R. The synergistic effect of radiotherapy and immunotherapy: A promising but not simple partnership. Crit. Rev. Oncol. Hematol. 2017;111:124–132. doi: 10.1016/j.critrevonc.2017.01.017.
- De Ruysscher D. Combination of radiotherapy and immune treatment: First clinical data. Cancer Radiother. 2018;22:564–566. doi: 10.1016/j.canrad.2018.07.128.
- Formenti S.C., Demaria S. Combining radiotherapy and cancer immunotherapy: A paradigm shift. J. Natl. Cancer Inst. 2013;105:256–265. doi: 10.1093/jnci/djs629.
- Sharma A., Bode B., Wenger R.H., Lehmann K., Sartori A.A., Moch H., Knuth A., Boehmer L., Broek M. Gamma-Radiation promotes immunological recognition of cancer cells through increased expression of cancer-testis antigens in vitro and in vivo. PLoS ONE. 2011;6:e28217. doi: 10.1371/journal.pone.0028217.
- Malamas A.S., Gameiro S.R., Knudson K.M., Hodge J.W. Sublethal exposure to alpha radiation (223Ra dichloride) enhances various carcinomas’ sensitivity to lysis by antigen-specific cytotoxic T lymphocytes through calreticulin-mediated immunogenic modulation. Oncotarget. 2016;7:86937–86947. doi: 10.18632/oncotarget.13520.
- Garnett C.T., Palena C., Chakraborty M., Tsang K.Y., Schlom J., Hodge J.W. Sublethal irradiation of human tumor cells modulates phenotype resulting in enhanced killing by cytotoxic T lymphocytes. Cancer Res. 2004;64:7985–7994. doi: 10.1158/0008-5472.CAN-04-1525.
- Rock K.L., Reits E., Neefjes J. Present yourself! By MHC class I and MHC class II molecules. Trends Immunol. 2016;37:724–737. doi: 10.1016/j.it.2016.08.010.
- Axelrod M.L., Cook R.S., Johnson D.B., Balko J.M. Biological consequences of MHC-II expression by tumor cells in cancer. Clin. Cancer Res. 2019;25:2392–2402. doi: 10.1158/1078-0432.CCR-18-3200.
- Accolla R.S., Lombardo L., Abdallah R., Raval G., Forlani G., Tosi G. Boosting the MHC class II-restricted tumor antigen presentation to CD4+ T helper cells: A critical issue for triggering protective immunity and re-orienting the tumor microenvironment toward an anti-tumor state. Front. Oncol. 2014;4:32. doi: 10.3389/fonc.2014.00032.
- Karamooz E., Harriff M.J., Lewinsohn D.M. MR1-dependent antigen presentation. Semin. Cell Dev. Biol. 2018;84:58–64. doi: 10.1016/j.semcdb.2017.11.028.
- Crowther M.D., Dolton G., Legut M., Caillaud M.E., Lloyd A., Attaf M., Galloway S.A.E., Rius C., Farrell C.P., Szomolay B., et al. Genome-wide CRISPR-Cas9 screening reveals ubiquitous T cell cancer targeting via the monomorphic MHC class I-related protein MR1. Nat. Immunol. 2020;21:178–185. doi: 10.1038/s41590-019-0578-8.
- Gettinger S., Choi J., Hastings K., Truini A., Datar I., Sowell R., Wurtz A., Dong W., Cai G., Melnick M.A., et al. Impaired HLA class I antigen processing and presentation as a mechanism of acquired resistance to immune checkpoint inhibitors in lung cancer. Cancer Discov. 2017;7:1420–1435. doi: 10.1158/-17-0593.
- Seliger B., Maeurer M.J., Ferrone S. Antigen-processing machinery breakdown and tumor growth. Immunol. Today. 2000;21:455–464. doi: 10.1016/S0167-5699(00)01692-3.
- Wang S., He Z., Wang X., Li H., Liu X.S. Antigen presentation and tumor immunogenicity in cancer immunotherapy response prediction. Elife. 2019;8 doi: 10.7554/eLife.49020.
- Chiriva-Internati M., Grizzi F., Pinkston J., Morrow K.J., D’Cunha N., Frezza E.E., Muzzio P.C., Kast W.M., Cobos E. Gamma-radiation upregulates MHC class I/II and ICAM-I molecules in multiple myeloma cell lines and primary tumors. Cell. Dev. Biol. Anim. 2006;42:89–95. doi: 10.1290/0508054.1.
- Reits E.A., Hodge J.W., Herberts C.A., Groothuis T.A., Chakraborty M., Wansley E.K., Camphausen K., Luiten R.M., De Ru A.H., Neijssen J., et al. Radiation modulates the peptide repertoire, enhances MHC class I expression, and induces successful antitumor immunotherapy. J. Exp. Med. 2006;203:1259–1271. doi: 10.1084/jem.20052494.
- Santin A.D., Hermonat P.L., Hiserodt J.C., Chiriva-Internati M., Woodliff J., Theus J.W., Barclay D., Pecorelli S., Parham G.P. Effects of irradiation on the expression of major histocompatibility complex class I antigen and adhesion costimulation molecules ICAM-1 in human cervical cancer. Int. J. Radiat. Oncol. Biol. Phys. 1997;39:737–742. doi: 10.1016/S0360-3016(97)00372-6.
- Larkins B.A., Dilkes B.P., Dante R.A., Coelho C.M., Woo Y.M., Liu Y. Investigating the hows and whys of DNA endoreduplication. J. Exp. Bot. 2001;52:183–192. doi: 10.1093/jexbot/52.355.183.
- Kaur E., Rajendra J., Jadhav S., Shridhar E., Goda J.S., Moiyadi A., Dutt S. Radiation-induced homotypic cell fusions of innately resistant glioblastoma cells mediate their sustained survival and recurrence. Carcinogenesis. 2015;36:685–695. doi: 10.1093/carcin/bgv050.
- Mirzayans R., Andrais B., Scott A., Wang Y.W., Kumar P., Murray D. Multinucleated giant cancer cells produced in response to ionizing radiation retain viability and replicate their genome. Int. J. Mol. Sci. 2017;18:360. doi: 10.3390/ijms18020360.
- Erenpreisa J., Ivanov A., Wheatley S.P., Kosmacek E.A., Ianzini F., Anisimov A.P., Mackey M., Davis P.J., Plakhins G., Illidge T.M. Endopolyploidy in irradiated p53-deficient tumour cell lines: Persistence of cell division activity in giant cells expressing Aurora-B kinase. Cell Biol. Int. 2008;32:1044–1056. doi: 10.1016/j.cellbi.2008.06.003.
- Schwarz-Finsterle J., Scherthan H., Huna A., Gonzalez P., Mueller P., Schmitt E., Erenpreisa J., Hausmann M. Volume increase and spatial shifts of chromosome territories in nuclei of radiation-induced polyploidizing tumour cells. Mutat. Res. 2013;756:56–65. doi: 10.1016/j.mrgentox.2013.05.004.
- Rene A.A., Nardone R.M. The effect of gamma radiation on cell enlargement and size distribution of strain L. Curr. Mod. Biol. 1968;2:207–214. doi: 10.1016/0303-2647(68)90005-1.
- Bairoch A. The cellosaurus, a cell-line knowledge resource. J. Biomol. Tech. 2018;29:25–38. doi: 10.7171/jbt.18-2902-002.
- Scholtalbers J., Boegel S., Bukur T., Byl M., Goerges S., Sorn P., Loewer M., Sahin U., Castle J.C. TCLP: An online cancer cell line catalogue integrating HLA type, predicted neo-epitopes, virus and gene expression. Genome Med. 2015;7:118. doi: 10.1186/s13073-015-0240-5.
- Jung M., Ramankulov A., Roigas J., Johannsen M., Ringsdorf M., Kristiansen G., Jung K. In search of suitable reference genes for gene expression studies of human renal cell carcinoma by real-time PCR. BMC Mol. Biol. 2007;8:47. doi: 10.1186/1471-2199-8-47.
- Hamann M.V., Mullers E., Reh J., Stanke N., Effantin G., Weissenhorn W., Lindemann D. The cooperative function of arginine residues in the Prototype Foamy Virus Gag C-terminus mediates viral and cellular RNA encapsidation. Retrovirology. 2014;11:87. doi: 10.1186/s12977-014-0087-7.
- Meissner T.B., Li A., Biswas A., Lee K.H., Liu Y.J., Bayir E., Iliopoulos D., Van den Elsen P.J., Kobayashi K.S. NLR family member NLRC5 is a transcriptional regulator of MHC class I genes. Proc. Natl. Acad. Sci. USA. 2010;107:13794–13799. doi: 10.1073/pnas.1008684107.
- Wang X., Xiong L., Yu G., Li D., Peng T., Luo D., Xu J. Cathepsin S silencing induces apoptosis of human hepatocellular carcinoma cells. Am. J. Transl. Res. 2015;7:100–110.
- Bolger A.M., Lohse M., Usadel B. Trimmomatic: A flexible trimmer for Illumina sequence data. Bioinformatics. 2014;30:2114–2120. doi: 10.1093/bioinformatics/btu170.
- Kim D., Langmead B., Salzberg S.L. HISAT: A fast spliced aligner with low memory requirements. Nat. Methods. 2015;12:357–360. doi: 10.1038/nmeth.3317.
- Liao Y., Smyth G.K., Shi W. The Subread aligner: Fast, accurate and scalable read mapping by seed-and-vote. Nucleic Acids Res. 2013;41:e108. doi: 10.1093/nar/gkt214.
- Robinson M.D., McCarthy D.J., Smyth G.K. edgeR: A bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics. 2010;26:139–140. doi: 10.1093/bioinformatics/btp616.
- Huang D.W., Sherman B.T., Lempicki R.A. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat. Protoc. 2009;4:44–57. doi: 10.1038/nprot.2008.211.
- Smyth G. Limma: Linear models for microarray data. In: Gentleman R., Carey V.J., Huber W., Dudoit S., Irizarry R.A., editors. Bioinformatics and Computational Biology Solutions using R and Bioconductor. Springer; New York, NY, USA: 2005. pp. 397–420.
- Almeida L.G., Sakabe N.J., de Oliveira A.R., Silva M.C., Mundstein A.S., Cohen T., Chen Y.T., Chua R., Gurung S., Gnjatic S., et al. CTdatabase: A knowledge-base of high-throughput and curated data on cancer-testis antigens. Nucleic Acids Res. 2009;37:D816–D819. doi: 10.1093/nar/gkn673.
Source: PubMed