Abstract
Background
Gastric cancer (GC) is one of the most prevalent types of cancer worldwide. B7-H3, an immune checkpoint molecule with promising potential, has been found to be overexpressed in various cancers. CD47 is an anti-phagocytic molecule that interacts with the signal regulatory protein alpha (SIRPα) to affect phagocytes. The relationship between the expression of B7-H3 and CD47, two potential therapeutic targets found in tumor cells, remains unknown. In this study, our objective is to investigate the clinical significance of co-expression of B7-H3 and CD47, as well as the potential therapeutic value of combination therapy in GC.
Methods
We utilized immunohistochemistry (IHC) to assess the expression of B7-H3, CD47, CD68, CD86 and CD163 in tissue microarrays obtained from 268 GC patients who underwent surgeries. Western blotting was employed to assess the protein level of B7-H3 and CD47 in GC tissues. The co-localization of B7-H3/CD47 and CD68 in GC tissues was determined using multiplex immunohistochemistry (m-IHC). We further verified the relationship between B7-H3/CD47 and macrophage infiltration via flow cytometry. To estimate the clinical outcomes of patients from different subgroups, we employed the Kaplan–Meier curve and the Cox model.
Results
Among the 268 GC cases, a total of 180 cases exhibited positive expression of B7-H3, while 122 cases showed positive expression of CD47. In fresh GC clinical tissues, B7-H3 and CD47 protein level was also higher in tumor tissue than in adjacent normal tissue. Remarkably, 91 cases demonstrated co-expression of B7-H3 and CD47. We observed a significant correlation between B7-H3 expression and tumor stage (P = 0.001), differentiation (P = 0.045), and depth (P = 0.003). Additionally, there was a significant association between B7-H3 and CD47 expression (P = 0.018). The percentage of B7-H3 and CD47 double positive cells in fresh GC tumor tissues were elevated compared with control adjacent tissues regardless of CD45− or CD45+ cells (P = 0.0029, P = 0.0012). Patients with high B7-H3 or CD47 expression had significantly lower overall survival (OS) rates compared to those with low expression levels (P = 0.0176 or P = 0.0042). Surprisingly, patients with combined high expression of B7-H3 and CD47 exhibited a considerably worse prognosis than others (P = 0.0007). Univariate analysis revealed that cases with high expression of B7-H3, CD47, or both had significantly higher hazard ratios (HR) than cases with low expression of these markers. Furthermore, the results of multivariate analysis indicated that B7-H3/CD47 co-expression and CD47 expression alone are independent prognostic factors for overall survival. Moreover, significant correlations were observed between B7-H3 and CD68 expression, CD47 and CD68 expression, as well as B7-H3/CD47 co-expression and CD68 expression in GC patients (P < 0.001, P = 0.003, and P < 0.001). Flow cytometry test showed that the percentage of CD68-positive cells but not CD86-positive cells among B7-H3-positive or CD47-positive immune cells in GC tumor tissue was elevated significantly compared with adjacent tissue.
Conclusion
Our findings demonstrated a correlation between B7-H3 expression and CD47 expression in GC patient tissues. Co-expression of B7-H3 and CD47 can serve as an indicator of poor prognosis in GC patients. In GC tumor tissue, but not adjacent tissue, B7-H3 and CD47 expression was accompanied with macrophage infiltration.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
All data used in this study are included in the article. Please contact the corresponding author for data requests.
References
Baccelli I, Stenzinger A, Vogel V et al (2014) Co-expression of MET and CD47 is a novel prognosticator for survival of luminal breast cancer patients. Oncotarget 5:8147–8160. https://doi.org/10.18632/oncotarget.2385
Barrera L, Montes-Servín E, Hernandez-Martinez J-M et al (2017) CD47 overexpression is associated with decreased neutrophil apoptosis/phagocytosis and poor prognosis in non-small-cell lung cancer patients. Br J Cancer 117:385–397. https://doi.org/10.1038/bjc.2017.173
Brahmer JR, Tykodi SS, Chow LQM et al (2012) Safety and activity of anti-PD-L1 antibody in patients with advanced cancer. N Engl J Med 366:2455–2465. https://doi.org/10.1056/NEJMoa1200694
Brown EJ, Frazier WA (2001) Integrin-associated protein (CD47) and its ligands. Trends Cell Biol 11:130–135. https://doi.org/10.1016/s0962-8924(00)01906-1
Casey SC, Tong L, Li Y et al (2016) MYC regulates the antitumor immune response through CD47 and PD-L1. Science 352:227–231. https://doi.org/10.1126/science.aac9935
Chapoval AI, Ni J, Lau JS et al (2001) B7–H3: a costimulatory molecule for T cell activation and IFN-γ production. Nat Immunol 2:269–274. https://doi.org/10.1038/85339
Cheng N, Bei Y, Song Y et al (2021) B7–H3 augments the pro-angiogenic function of tumor-associated macrophages and acts as a novel adjuvant target for triple-negative breast cancer therapy. Biochem Pharmacol 183:114298. https://doi.org/10.1016/j.bcp.2020.114298
Feng M, Jiang W, Kim BYS et al (2019) Phagocytosis checkpoints as new targets for cancer immunotherapy. Nat Rev Cancer 19:568–586. https://doi.org/10.1038/s41568-019-0183-z
Feng R, Chen Y, Liu Y et al (2021) The role of B7–H3 in tumors and its potential in clinical application. Int Immunopharmacol 101:108153. https://doi.org/10.1016/j.intimp.2021.108153
Garon EB, Rizvi NA, Hui R et al (2015) Pembrolizumab for the treatment of non-small-cell lung cancer. N Engl J Med 372:2018–2028. https://doi.org/10.1056/NEJMoa1501824
Hayat SMG, Bianconi V, Pirro M et al (2020) CD47: role in the immune system and application to cancer therapy. Cell Oncol Dordr 43:19–30. https://doi.org/10.1007/s13402-019-00469-5
Haydar D, Houke H, Chiang J et al (2021) Cell-surface antigen profiling of pediatric brain tumors: B7–H3 is consistently expressed and can be targeted via local or systemic CAR T-cell delivery. Neuro-Oncol 23:999–1011. https://doi.org/10.1093/neuonc/noaa278
Hodi FS, O’Day SJ, McDermott DF et al (2010) Improved survival with ipilimumab in patients with metastatic melanoma. N Engl J Med 363:711–723. https://doi.org/10.1056/NEJMoa1003466
Huang Y-K, Wang M, Sun Y et al (2019) Macrophage spatial heterogeneity in gastric cancer defined by multiplex immunohistochemistry. Nat Commun 10:3928. https://doi.org/10.1038/s41467-019-11788-4
Imam R, Chang Q, Black M et al (2021) CD47 expression and CD163+ macrophages correlated with prognosis of pancreatic neuroendocrine tumor. BMC Cancer 21:320. https://doi.org/10.1186/s12885-021-08045-7
Ingebrigtsen VA, Boye K, Tekle C et al (2012) B7–H3 expression in colorectal cancer: nuclear localization strongly predicts poor outcome in colon cancer. Int J Cancer 131:2528–2536. https://doi.org/10.1002/ijc.27566
Kang F-B, Wang L, Li D et al (2015) Hepatocellular carcinomas promote tumor-associated macrophage M2-polarization via increased B7–H3 expression. Oncol Rep 33:274–282. https://doi.org/10.3892/or.2014.3587
Kontos F, Michelakos T, Kurokawa T et al (2021) B7–H3: an attractive target for antibody-based immunotherapy. Clin Cancer Res off J Am Assoc Cancer Res 27:1227–1235. https://doi.org/10.1158/1078-0432.CCR-20-2584
Li F, Lv B, Liu Y et al (2018) Blocking the CD47-SIRPα axis by delivery of anti-CD47 antibody induces antitumor effects in glioma and glioma stem cells. Oncoimmunology 7:e1391973. https://doi.org/10.1080/2162402X.2017.1391973
Mao Y, Chen L, Wang F et al (2017) Cancer cell-expressed B7–H3 regulates the differentiation of tumor-associated macrophages in human colorectal carcinoma. Oncol Lett 14:6177–6183. https://doi.org/10.3892/ol.2017.6935
McCracken MN, Cha AC, Weissman IL (2015) Molecular pathways: activating t cells after cancer cell phagocytosis from blockade of CD47 “don’t eat me” signals. Clin Cancer Res off J Am Assoc Cancer Res 21:3597–3601. https://doi.org/10.1158/1078-0432.CCR-14-2520
Miyamoto T, Murakami R, Hamanishi J et al (2022) B7–H3 suppresses antitumor immunity via the CCL2–CCR2–M2 macrophage axis and contributes to ovarian cancer progression. Cancer Immunol Res 10:56–69. https://doi.org/10.1158/2326-6066.CIR-21-0407
Nishida T, Sugimoto A, Tomita R et al (2019) Impact of time from diagnosis to chemotherapy in advanced gastric cancer: a propensity score matching study to balance prognostic factors. World J Gastrointest Oncol 11:28–38. https://doi.org/10.4251/wjgo.v11.i1.28
Okazawa H, Motegi S, Ohyama N et al (2005) Negative regulation of phagocytosis in macrophages by the CD47-SHPS-1 system. J Immunol Baltim Md 174:2004–2011. https://doi.org/10.4049/jimmunol.174.4.2004
Oldenborg PA, Gresham HD, Lindberg FP (2001) CD47-signal regulatory protein alpha (SIRPalpha) regulates Fcgamma and complement receptor-mediated phagocytosis. J Exp Med 193:855–862. https://doi.org/10.1084/jem.193.7.855
Picarda E, Ohaegbulam KC, Zang X (2016) Molecular pathways: targeting B7–H3 (CD276) for human cancer immunotherapy. Clin Cancer Res off J Am Assoc Cancer Res 22:3425. https://doi.org/10.1158/1078-0432.CCR-15-2428
Prasad DVR, Nguyen T, Li Z et al (2004) Murine B7–H3 Is a negative regulator of T cells. J Immunol 173:2500–2506. https://doi.org/10.4049/jimmunol.173.4.2500
Purvis IJ, Avilala J, Guda MR et al (2019) Role of MYC-miR-29-B7-H3 in Medulloblastoma growth and angiogenesis. J Clin Med 8:1158. https://doi.org/10.3390/jcm8081158
Qian B-Z, Pollard JW (2010) Macrophage diversity enhances tumor progression and metastasis. Cell 141:39–51. https://doi.org/10.1016/j.cell.2010.03.014
Shao X, Zhan S, Quan Q et al (2022) Clinical significance of B7–H3 and HER2 co-expression and therapeutic value of combination treatment in gastric cancer. Int Immunopharmacol 110:108988. https://doi.org/10.1016/j.intimp.2022.108988
Shi M, Gu Y, Jin K et al (2021) CD47 expression in gastric cancer clinical correlates and association with macrophage infiltration. Cancer Immunol Immunother 70:1831–1840. https://doi.org/10.1007/s00262-020-02806-2
Sikic BI, Lakhani N, Patnaik A et al (2019) First-in-human, first-in-class phase I trial of the anti-CD47 antibody Hu5F9-G4 in patients with advanced cancers. J Clin Oncol off J Am Soc Clin Oncol 37:946–953. https://doi.org/10.1200/JCO.18.02018
Smyth EC, Nilsson M, Grabsch HI et al (2020) Gastric cancer. Lancet Lond Engl 396:635–648. https://doi.org/10.1016/S0140-6736(20)31288-5
Sockolosky JT, Dougan M, Ingram JR et al (2016) Durable antitumor responses to CD47 blockade require adaptive immune stimulation. Proc Natl Acad Sci 113:E2646–E2654. https://doi.org/10.1073/pnas.1604268113
Suh W-K, Gajewska BU, Okada H et al (2003) The B7 family member B7–H3 preferentially down-regulates T helper type 1–mediated immune responses. Nat Immunol 4:899–906. https://doi.org/10.1038/ni967
Sung H, Ferlay J, Siegel RL et al (2021) Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 71:209–249. https://doi.org/10.3322/caac.21660
Tan M, Zhu L, Zhuang H et al (2015) Lewis Y antigen modified CD47 is an independent risk factor for poor prognosis and promotes early ovarian cancer metastasis. Am J Cancer Res 5:2777–2787
Topalian SL, Hodi FS, Brahmer JR et al (2012) Safety, activity, and immune correlates of anti-PD-1 antibody in cancer. N Engl J Med 366:2443–2454. https://doi.org/10.1056/NEJMoa1200690
Wu C-P, Jiang J-T, Tan M et al (2006) Relationship between co-stimulatory molecule B7–H3 expression and gastric carcinoma histology and prognosis. World J Gastroenterol 12:457–459. https://doi.org/10.3748/wjg.v12.i3.457
Wu L, Yu G-T, Deng W-W et al (2018) Anti-CD47 treatment enhances anti-tumor T-cell immunity and improves immunosuppressive environment in head and neck squamous cell carcinoma. Oncoimmunology 7:e1397248. https://doi.org/10.1080/2162402X.2017.1397248
Ye Z, Zheng Z, Li X et al (2016) B7–H3 Overexpression predicts poor survival of cancer patients: a meta-analysis. Cell Physiol Biochem Int J Exp Cell Physiol Biochem Pharmacol 39:1568–1580. https://doi.org/10.1159/000447859
Yoshida K, Tsujimoto H, Matsumura K et al (2015) CD47 is an adverse prognostic factor and a therapeutic target in gastric cancer. Cancer Med 4:1322–1333. https://doi.org/10.1002/cam4.478
Yu L, Ding Y, Wan T et al (2021) Significance of CD47 and its association with tumor immune microenvironment heterogeneity in ovarian cancer. Front Immunol 12:768115. https://doi.org/10.3389/fimmu.2021.768115
Yunna C, Mengru H, Lei W, Weidong C (2020) Macrophage M1/M2 polarization. Eur J Pharmacol 877:173090. https://doi.org/10.1016/j.ejphar.2020.173090
Zang X, Thompson RH, Al-Ahmadie HA et al (2007) B7–H3 and B7x are highly expressed in human prostate cancer and associated with disease spread and poor outcome. Proc Natl Acad Sci USA 104:19458–19463. https://doi.org/10.1073/pnas.0709802104
Zang X, Sullivan PS, Soslow RA et al (2010) Tumor associated endothelial expression of B7–H3 predicts survival in ovarian carcinomas. Mod Pathol 23:1104–1112. https://doi.org/10.1038/modpathol.2010.95
Zhan S, Liu Z, Zhang M et al (2020) Overexpression of B7–H3 in α-SMA-positive fibroblasts is associated with cancer progression and survival in gastric adenocarcinomas. Front Oncol 9:1466. https://doi.org/10.3389/fonc.2019.01466
Zhang X, Fang C, Zhang G et al (2017) Prognostic value of B7–H3 expression in patients with solid tumors: a meta-analysis. Oncotarget 8:93156–93167. https://doi.org/10.18632/oncotarget.21114
Zhao B, Li H, Xia Y et al (2022) Immune checkpoint of B7–H3 in cancer: from immunology to clinical immunotherapy. J Hematol OncolJ Hematol Oncol 15:153. https://doi.org/10.1186/s13045-022-01364-7
Funding
This study was fund by grants of Suzhou Science and Technology Development Plan (SKY2022129) and the National Natural Science Foundation of China (Grant no. 81874163 and 82001723).
Author information
Authors and Affiliations
Contributions
All authors contributed to the study conception and design. SC: Writing-original draft, Statistical analysis. SZ: Data curation, Resources. SD: Resources, Funding acquisition. QZ: Data collection. HX: Material preparation. XZ: Supervision. LC: Supervision, Data analysis, Writing-review and editing, Funding acquisition. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest that could influence the submitted work.
Ethical approval and consent to participate
This retrospective study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the Ethics Committee of Soochow University (approval number: 2018121).
Informed consent
Since this study was a retrospective study, the Ethics Committee of The First Affiliated Hospital of Soochow University has confirmed that no informed consent is required.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
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.
About this article
Cite this article
Chen, S., Zhan, S., Ding, S. et al. B7-H3 and CD47 co-expression in gastric cancer is a predictor of poor prognosis and potential targets for future dual-targeting immunotherapy. J Cancer Res Clin Oncol 149, 16609–16621 (2023). https://doi.org/10.1007/s00432-023-05408-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00432-023-05408-4