Systematic characterization of the tumor microenvironment in Chinese patients with hepatocellular carcinoma highlights intratumoral B cells as a potential immunotherapy target

Feng,Yu; Liu,Liguo; Li,Jing; Huang,Jia; Xie,Jenny H.; Menard,Laurence; Shi,Yanfen; Zhao,Xiaohong; Xie,Shan; Zang,Wenjuan; Tan,Haidong; Yang,Zhiying; Ni,Ling

doi:10.3892/or.2021.8249

Systematic characterization of the tumor microenvironment in Chinese patients with hepatocellular carcinoma highlights intratumoral B cells as a potential immunotherapy target

Authors:
- Yu Feng
- Liguo Liu
- Jing Li
- Jia Huang
- Jenny H. Xie
- Laurence Menard
- Yanfen Shi
- Xiaohong Zhao
- Shan Xie
- Wenjuan Zang
- Haidong Tan
- Zhiying Yang
- Ling Ni
View Affiliations

Affiliations: Institute for Immunology and School of Medicine, Tsinghua University, Beijing 100084, P.R. China, Department of Hepatobiliary Surgery, China‑Japan Friendship Hospital, Beijing 100029, P.R. China, Research and Early Development, Bristol Myers Squibb, Princeton, NJ 08540, USA, Department of Pathology, China‑Japan Friendship Hospital, Beijing 100029, P.R. China
Published online on: December 24, 2021 https://doi.org/10.3892/or.2021.8249
Article Number: 38
Copyright: © Feng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Hepatocellular carcinoma (HCC) is an immunogenic malignancy, which exhibits low responsiveness to programmed cell death protein‑1 (PD‑1)/programmed death ligand‑1 (PD‑L1) antibodies. Therefore, the identification of novel immunotherapeutic targets to treat HCC is imperative. Systematic characterization of the HCC tumor microenvironment (TME) can provide novel insight into additional therapeutic approaches. In the present study, the RNA‑sequencing (RNA‑seq) data of 360 patients with HCC were integrated from The Cancer Genome Atlas to assess the expression of membrane spanning 4‑domains A1 (MS4A1; encoding CD20) in tumors and normal liver tissues. Immunofluorescence and multiplex tissue fluorescence analyses were performed and combined with flow cytometry staining to measure CD20/CD19 expression at the protein level. In addition, published single cell RNA‑seq data of CD45⁺ cells were derived from 16 treatment‑naïve patients from Beijing Shijitan Hospital with HCC to illustrate the characteristics of CD19⁺ B cells. The results indicated that the HCC TME included nuclear receptor subfamily 4 group A member 2⁺ (NR4A2) B cells. Patients with HCC and high density of intratumoral B cells demonstrated compromised antitumor immunity manifested by low percentages of cytotoxic CD8⁺ T cells and high density of regulatory T cells. Furthermore, PD‑L1 expression was significantly correlated with the B cell signature marker CD20. The present study indicated that tumor‑infiltrating B cells may play a negative role in antitumor immunity and serve as a promising target for HCC immunotherapy, alone or in combination with anti‑PD‑L1/PD‑1 antibodies.

View Figures

View References

1	Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D and Bray F: Cancer incidence and mortality worldwide: Sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer. 136:E359–E386. 2015. View Article : Google Scholar : PubMed/NCBI
2	European Association for the Study of the Liver. Electronic address, . simpleeasloffice@easloffice.eu; European Association for the Study of the Liver: EASL clinical practice guidelines: Management of hepatocellular carcinoma. J Hepatol. 69:182–236. 2018. View Article : Google Scholar : PubMed/NCBI
3	Zhu RX, Seto WK, Lai CL and Yuen MF: Epidemiology of hepatocellular carcinoma in the asia-pacific region. Gut Liver. 10:332–339. 2016. View Article : Google Scholar : PubMed/NCBI
4	Yang JD and Roberts LR: Hepatocellular carcinoma: A global view. Nat Rev Gastroenterol Hepatol. 7:448–458. 2010. View Article : Google Scholar : PubMed/NCBI
5	Llovet JM, Ricci S, Mazzaferro V, Hilgard P, Gane E, Blanc JF, de Oliveira AC, Santoro A, Raoul JL, Forner A, et al: Sorafenib in advanced hepatocellular carcinoma. N Engl J Med. 359:378–390. 2008. View Article : Google Scholar : PubMed/NCBI
6	Buonaguro L, Mauriello A, Cavalluzzo B, Petrizzo A and Tagliamonte M: Immunotherapy in hepatocellular carcinoma. Ann Hepatol. 18:291–297. 2019. View Article : Google Scholar : PubMed/NCBI
7	El-Khoueiry AB, Sangro B, Yau T, Crocenzi TS, Kudo M, Hsu C, Kim TY, Choo SP, Trojan J, Rd THW, et al: Nivolumab in patients with advanced hepatocellular carcinoma (CheckMate 040): An open-label, non-comparative, phase 1/2 dose escalation and expansion trial. Lancet. 389:2492–2502. 2017. View Article : Google Scholar : PubMed/NCBI
8	Yarchoan M, Hopkins A and Jaffee EM: Tumor mutational burden and response rate to PD-1 inhibition. N Engl J Med. 377:2500–2501. 2017. View Article : Google Scholar : PubMed/NCBI
9	Leonardi GC, Candido S, Cervello M, Nicolosi D, Raiti F, Travali S, Spandidos DA and Libra M: The tumor microenvironment in hepatocellular carcinoma (review). Int J Oncol. 40:1733–1747. 2012.PubMed/NCBI
10	Shalapour S, Lin XJ, Bastian IN, Brain J, Burt AD, Aksenov AA, Vrbanac AF, Li W, Perkins A, Matsutani T, et al: Inflammation-induced IgA⁺ cells dismantle anti-liver cancer immunity. Nature. 551:340–345. 2017. View Article : Google Scholar : PubMed/NCBI
11	Ye L, Zhang Q, Cheng Y, Chen X, Wang G, Shi M, Zhang T, Cao Y, Pan H, Zhang L, et al: Tumor-derived exosomal HMGB1 fosters hepatocellular carcinoma immune evasion by promoting TIM-1(+) regulatory B cell expansion. J Immunother Cancer. 6:1452018. View Article : Google Scholar : PubMed/NCBI
12	Xue H, Lin F, Tan H, Zhu ZQ, Zhang ZY and Zhao L: Overrepresentation of IL-10-expressing B cells suppresses cytotoxic CD4⁺ T cell activity in HBV-induced hepatocellular carcinoma. PLoS One. 11:e01548152016. View Article : Google Scholar : PubMed/NCBI
13	Schwartz M, Zhang Y and Rosenblatt JD: B cell regulation of the anti-tumor response and role in carcinogenesis. J Immunother Cancer. 4:402016. View Article : Google Scholar : PubMed/NCBI
14	Shaso Y, Lo CM, Ling CC, Liu XB, Ng KTP, Chu AC, Ma YY, Li CX, Fan ST and Man K: Regulatory B cells accelerate hepatocellular carcinoma progression via CD40/CD154 signaling pathway. Cancer Lett. 355:264–272. 2014. View Article : Google Scholar : PubMed/NCBI
15	Zhang Z, Ma L, Goswami S, Ma J, Zheng B, Duan M, Liu L, Zhang L, Shi J, Dong L, et al: Landscape of infiltrating B cells and their clinical significance in human hepatocellular carcinoma. Oncoimmunology. 8:e15713882019. View Article : Google Scholar : PubMed/NCBI
16	Garnelo M, Tan A, Her Z, Yeong J, Lim CJ, Chen J, Lim KH, Weber A, Chow P, Chung A, et al: Interaction between tumour-infiltrating B cells and T cells controls the progression of hepatocellular carcinoma. Gut. 66:342–351. 2017. View Article : Google Scholar : PubMed/NCBI
17	Cabrita R, Lauss M, Sanna A, Donia M, Larsen MS, Mitra S, Johansson I, Phung B, Harbst K, Vallon-Christersson J, et al: Tertiary lymphoid structures improve immunotherapy and survival in melanoma. Nature. 577:561–565. 2020. View Article : Google Scholar : PubMed/NCBI
18	Helmink BA, Reddy SM, Gao J, Zhang S, Basar R, Thakur R, Yizhak K, Sade-Feldman M, Blando J, Han G, et al: B cells and tertiary lymphoid structures promote immunotherapy response. Nature. 577:549–555. 2020. View Article : Google Scholar : PubMed/NCBI
19	Petitprez F, de Reynies A, Keung EZ, Chen TWW, Sun CM, Calderaro J, Jeng YM, Hsiao LP, Lacroix L, Bougoüin A, et al: B cells are associated with survival and immunotherapy response in sarcoma. Nature. 577:556–560. 2020. View Article : Google Scholar : PubMed/NCBI
20	Xie S, Huang J, Qiao Q, Zang W, Hong S, Tan H, Dong C, Yang Z and Ni L: Expression of the inhibitory B7 family molecule VISTA in human colorectal carcinoma tumors. Cancer Immunol Immunother. 67:1685–1694. 2018. View Article : Google Scholar : PubMed/NCBI
21	Stuart T, Butler A, Hoffman P, Hafemeister C, Papalexi E, Mauck WM III, Hao Y, Stoeckius M, Smibert P and Satija R: Comprehensive integration of single-cell data. Cell. 177:1888–1902.e21. 2019. View Article : Google Scholar : PubMed/NCBI
22	McInne L and Healy J: UMAP: Uniform Manifold Approximation and Projection for Dimension Reduction. ArXiv e-prints 1802.03426. 2018.https://arxiv.org/abs/1802.03426
23	Zhang Q, He Y, Luo N, Patel SJ, Han Y, Gao R, Modak M, Carotta S, Haslinger C, Kind D, et al: Landscape and dynamics of single immune cells in hepatocellular carcinoma. Cell. 179:829–845, e820. 2019. View Article : Google Scholar : PubMed/NCBI
24	Li QX, Ke N, Sundaram R and Wong-Staal F: NR4A1, 2, 3-an orphan nuclear hormone receptor family involved in cell apoptosis and carcinogenesis. Histol Histopathol. 21:533–540. 2006.PubMed/NCBI
25	Liu X, Wang Y, Lu H, Li J, Yan X, Xiao M, Hao J, Alekseev A, Khong H, Chen T, et al: Genome-wide analysis identifies NR4A1 as a key mediator of T cell dysfunction. Nature. 567:525–529. 2019. View Article : Google Scholar : PubMed/NCBI
26	Chen J, Lopez-Moyado IF, Seo H, Lio CWJ, Hempleman LJ, Sekiya T, Yoshimura A, Scott-Browne JP and Rao A: NR4A transcription factors limit CAR T cell function in solid tumours. Nature. 567:530–534. 2019. View Article : Google Scholar : PubMed/NCBI
27	Sakaguchi S, Yamaguchi T, Nomura T and Ono M: Regulatory T cells and immune tolerance. Cell. 133:775–787. 2008. View Article : Google Scholar : PubMed/NCBI
28	Ren Z, Peng H and Fu YX: PD-1 shapes B cells as evildoers in the tumor microenvironment. Cancer Discov. 6:477–478. 2016. View Article : Google Scholar : PubMed/NCBI
29	de Visser KE, Korets LV and Coussens LM: De novo carcinogenesis promoted by chronic inflammation is B lymphocyte dependent. Cancer Cell. 7:411–423. 2005. View Article : Google Scholar : PubMed/NCBI
30	Andreu P, Johansson M, Affara NI, Pucci F, Tan T, Junankar S, Korets L, Lam J, Tawfik D, DeNardo DG, et al: FcRγ activation regulates inflammation-associated squamous carcinogenesis. Cancer Cell. 17:121–134. 2010. View Article : Google Scholar : PubMed/NCBI
31	Qin Z, Richter G, Schüler T, Ibe S, Cao X and Blankenstein T: B cells inhibit induction of T cell-dependent tumor immunity. Nat Med. 4:627–630. 1998. View Article : Google Scholar : PubMed/NCBI
32	Hetta HF: Regulatory B cells: Key players in hepatocellular carcinoma progression. Gastroenterology & Hepatology. Open Access 52016. https://medcraveonline.com/GHOA/GHOA-05-00136.pdf
33	DiLillo DJ, Matsushita T and Tedder TF: B10 cells and regulatory B cells balance immune responses during inflammation, autoimmunity, and cancer. Ann NY Acad Sci. 1183:38–57. 2010. View Article : Google Scholar : PubMed/NCBI
34	Shevach EM: From vanilla to 28 flavors: Multiple varieties of T regulatory cells. Immunity. 25:195–201. 2006. View Article : Google Scholar : PubMed/NCBI
35	von Boehmer H: Mechanisms of suppression by suppressor T cells. Nat Immunol. 6:338–344. 2005. View Article : Google Scholar : PubMed/NCBI
36	Lin CY, Graca L, Cobbold SP and Waldmann H: Dominant transplantation tolerance impairs CD8⁺ T cell function but not expansion. Nat Immunol. 3:1208–1213. 2002. View Article : Google Scholar : PubMed/NCBI