Serum BRD2 autoantibody in hepatocellular carcinoma and its detection using mimotope peptide‑conjugated BSA

Heo,Chang-Kyu; Lim,Won-Hee; Park,Inseo; Choi,Yon-Sik; Lim,Kook-Jin; Cho,Eun-Wie

doi:10.3892/ijo.2022.5448

Serum BRD2 autoantibody in hepatocellular carcinoma and its detection using mimotope peptide‑conjugated BSA

Authors:
- Chang-Kyu Heo
- Won-Hee Lim
- Inseo Park
- Yon-Sik Choi
- Kook-Jin Lim
- Eun-Wie Cho
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Affiliations: Rare Disease Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 34141, Republic of Korea, ProteomeTech Inc., Seoul 07528, Republic of Korea
Published online on: November 1, 2022 https://doi.org/10.3892/ijo.2022.5448
Article Number: 158
Copyright: © Heo et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Tumor‑associated (TA) autoantibodies are considered to be promising biomarkers for the early detection of cancer, prior to the development of clinical symptoms. In the present study, a novel TA autoantibody was detected, which may prove to be useful as a diagnostic marker of human HCC using an HBx‑transgenic (HBx‑tg) hepatocellular carcinoma (HCC) mouse model. Its target antigen was identified as the bromodomain‑containing protein 2 (BRD2), a transcriptional regulator that plays a pivotal role in the transcriptional control of diverse genes. BRD2 was upregulated in HCC tissues of the H‑ras12V‑tg mouse and human subjects, as demonstrated using western blotting or immunohistochemical analysis, with the BRD2 autoantibody. In addition, the truncated BRD2 reactive to the BRD2 autoantibody was detected in tumor cell‑derived exosomes, which possibly activated TA immune responses and the generation of autoantibodies. For the detection of the serum BRD2 autoantibody, epitope mimicries of autoantigenic BRD2 were screened from a random cyclic peptide CX₇C library with the BRD2 autoantibody. A mimotope with the sequence of CTSVFLPHC, which was cyclized by one pair of cysteine residues, exhibited high affinity to the BRD2 autoantibody and competitively inhibited the binding of the autoantibody to the cellular BRD2 antigen. The use of this cyclic peptide as a capture antigen in human serum enzyme‑linked immunosorbent assay allowed the distinction of patients with HCC from healthy subjects with 64.41% sensitivity and 82.42% specificity (area under the ROC curve, 0.7761), which is superior to serum alpha‑fetoprotein (AFP; 35.83% sensitivity; 100% specificity; area under the ROC curve, 0.5337) for the diagnosis of HCC. In addition, the detection of the BRD2 autoantibody combined with other autoantibody biomarkers or AFP has increased the accuracy of HCC diagnosis, suggesting that the combinational detection of cancer biomarkers, including the BRD2 autoantibody, is a promising assay for HCC diagnosis.

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1	World Health Organization (WHO): Cancer. WHO; Geneva: 2022, https://www.who.int/news-room/fact-sheets/detail/cancer. Accessed February 3, 2022.
2	Philips CA, Rajesh S, Nair DC, Ahamed R, Abduljaleel JK and Augustine P: Hepatocellular carcinoma in 2021: An exhaustive update. Cureus. 13:e192742021.PubMed/NCBI
3	Wong RJ, Cheung R and Ahmed A: Nonalcoholic steatohepatitis is the most rapidly growing indication for liver transplantation in patients with hepatocellular carcinoma in the U.S. Hepatology. 59:2188–2195. 2014. View Article : Google Scholar : PubMed/NCBI
4	Anstee QM, Reeves HL, Kotsiliti E, Govaere O and Heikenwalder M: From NASH to HCC: Current concepts and future challenges. Nat Rev Gastroenterol Hepatol. 16:411–428. 2019. View Article : Google Scholar : PubMed/NCBI
5	Forner A, Reig M and Bruix J: Hepatocellular carcinoma. Lancet. 391:1301–1314. 2018. View Article : Google Scholar : PubMed/NCBI
6	Hartke J, Johnson M and Ghabril M: The diagnosis and treatment of hepatocellular carcinoma. Semin Diagn Pathol. 34:153–159. 2017. View Article : Google Scholar : PubMed/NCBI
7	Marshall HT and Djamgoz MBA: Immuno-oncology: Emerging targets and combination therapies. Front Oncol. 8:3152018. View Article : Google Scholar : PubMed/NCBI
8	Vinay DS, Ryan EP, Pawelec G, Talib WH, Stagg J, Elkord E, Lichtor T, Decker WK, Whelan RL, Kumara HMCS, et al: Immune evasion in cancer: Mechanistic basis and therapeutic strategies. Semin Cancer Biol. 35(Suppl): S185–S198. 2015. View Article : Google Scholar : PubMed/NCBI
9	Sharonov GV, Serebrovskaya EO, Yuzhakova DV, Britanova OV and Chudakov DM: B cells, plasma cells and antibody repertoires in the tumour microenvironment. Nat Rev Immunol. 20:294–307. 2020. View Article : Google Scholar : PubMed/NCBI
10	Macdonald IK, Parsy-Kowalska CB and Chapman CJ: Autoantibodies: Opportunities for early cancer detection. Trends Cancer. 3:198–213. 2017. View Article : Google Scholar : PubMed/NCBI
11	Kobayashi M, Katayama H, Fahrmann JF and Hanash SM: Development of autoantibody signatures for common cancers. Semin Immunol. 47:1013882020. View Article : Google Scholar : PubMed/NCBI
12	de Jonge H, Iamele L, Maggi M, Pessino G and Scotti C: Anti-cancer auto-antibodies: Roles, applications and open issues. Cancers (Basel). 13:8132021. View Article : Google Scholar :
13	Sexauer D, Gray E and Zaenker P: Tumour-associated autoantibodies as prognostic cancer biomarkers-a review. Autoimmun Rev. 21:1030412022. View Article : Google Scholar
14	Wouters MCA and Nelson BH: Prognostic significance of tumor-infiltrating B cells and plasma cells in human cancer. Clin Cancer Res. 24:6125–6135. 2018. View Article : Google Scholar : PubMed/NCBI
15	Zheng M, Li YM, Liu ZY, Zhang X, Zhou Y, Jiang JL, Zhu P, Yang XM, Tang J and Chen ZN: Prognostic landscape of tumor-infiltrating T and B cells in human cancer. Front Immunol. 12:7313292022. View Article : Google Scholar : PubMed/NCBI
16	Heo CK, Hwang HM, Lim WH, Lee HJ, Yoo JS, Lim KJ and Cho EW: Cyclic peptide mimotopes for the detection of serum Anti-ATIC autoantibody biomarker in hepato-cellular carcinoma. Int J Mol Sci. 21:97182020. View Article : Google Scholar
17	Heo CK, Hwang HM, Lee HJ, Kwak SS, Yoo JS, Yu DY, Lim KJ, Lee S and Cho EW: Serum anti-EIF3A autoantibody as a potential diagnostic marker for hepatocellular carcinoma. Sci Rep. 9:110592019. View Article : Google Scholar : PubMed/NCBI
18	Hwang HM, Heo CK, Lee HJ, Kwak SS, Lim WH, Yoo JS, Yu DY, Lim KJ, Kim JY and Cho EW: Identification of anti-SF3B1 auto-antibody as a diagnostic marker in patients with hepatocellular carcinoma. J Transl Med. 16:1772018. View Article : Google Scholar
19	Zhang XD, Wang Y and Ye LH: Hepatitis B virus X protein accelerates the development of hepatoma. Cancer Biol Med. 11:182–190. 2014.PubMed/NCBI
20	Hsieh A, Kim HS, Lim SO, Yu DY and Jung G: Hepatitis B viral X protein interacts with tumor suppressor adenomatous polyposis coli to activate Wnt/β-catenin signaling. Cancer Lett. 300:162–172. 2011. View Article : Google Scholar
21	Yu DY, Moon HB, Son JK, Jeong S, Yu SL, Yoon H, Han YM, Lee CS, Park JS, Lee CH, et al: Incidence of hepatocellular carcinoma in transgenic mice expressing the hepatitis B virus X-protein. J Hepatol. 31:123–132. 1999. View Article : Google Scholar : PubMed/NCBI
22	National Library of Medicine (NIH): BRD2 bromodomain containing 2. NIH; Bethesda, MD: 2022, https://www.ncbi.nlm.nih.gov/gtr/genes/6046/. Updated October 9, 2022.
23	Loganathan SN, Tang N, Fleming JT, Ma Y, Guo Y, Borinstein SC, Chiang C and Wang J: BET bromodomain inhibitors suppress EWS-FLI1-dependent transcription and the IGF1 autocrine mechanism in Ewing sarcoma. Oncotarget. 7:43504–43517. 2016. View Article : Google Scholar : PubMed/NCBI
24	Sun LY, Spong A, Swindell WR, Fang Y, Hill C, Huber JA, Boehm JD, Westbrook R, Salvatori R and Bartke A: Growth hormone-releasing hormone disruption extends lifespan and regulates response to caloric restriction in mice. Elife. 2:e010982013. View Article : Google Scholar : PubMed/NCBI
25	Belkina AC, Blanton WP, Nikolajczyk BS and Denis GV: The double bromodomain protein Brd2 promotes B cell expansion and mitogenesis. J Leukoc Biol. 95:451–460. 2014. View Article : Google Scholar :
26	Pathak S, Stewart WCL, Burd CE, Hester ME and Greenberg DA: Brd2 haploinsufficiency extends lifespan and healthspan in C57B6/J mice. PLoS One. 15:e02349102020. View Article : Google Scholar : PubMed/NCBI
27	Hnilicova J, Hozeifi S, Stejskalova E, Duskova E, Poser I, Humpolickova J, Hof M and Staněk D: The C-terminal domain of Brd2 is important for chromatin interaction and regulation of transcription and alternative splicing. Mol Biol Cell. 24:3557–3568. 2013. View Article : Google Scholar : PubMed/NCBI
28	Suppiah A and Greenman J: Clinical utility of anti-p53 auto-antibody: Systematic review and focus on colorectal cancer. World J Gastroenterol. 19:4651–4670. 2013. View Article : Google Scholar : PubMed/NCBI
29	Zaenker P, Gray ES and Ziman MR: Autoantibody production in cancer-the humoral immune response toward autologous antigens in cancer patients. Autoimmun Rev. 15:477–483. 2016. View Article : Google Scholar : PubMed/NCBI
30	Kato T, Fahrmann JF, Hanash SM and Vykoukal J: Extracellular vesicles mediate B cell immune response and are a potential target for cancer therapy. Cells. 9:15182020. View Article : Google Scholar :
31	Hao Q, Wu Y, Wu Y, Wang P and Vadgama JV: Tumor-derived exosomes in tumor-induced immune suppression. Int J Mol Sci. 23:14612022. View Article : Google Scholar : PubMed/NCBI
32	Heo CK, Hwang HM, Ruem A, Yu DY, Lee JY, Yoo JS, Kim IG, Yoo HS, Oh S, Ko JH and Cho EW: Identification of a mimotope for circulating anti-cytokeratin 8/18 antibody and its usage for the diagnosis of breast cancer. Int J Oncol. 42:65–74. 2013. View Article : Google Scholar :
33	Heo CK, Woo MK, Yu DY, Lee JY, Yoo JS, Yoo HS, Ko JH, Kim JM, Choi JY, Kim IG, et al: Identification of autoantibody against fatty acid synthase in hepatocellular carcinoma mouse model and its application to diagnosis of HCC. Int J Oncol. 36:1453–1459. 2010.PubMed/NCBI
34	Wang T and Zhang KH: New blood biomarkers for the diagnosis of AFP-negative hepatocellular carcinoma. Front Oncol. 10:13162020. View Article : Google Scholar : PubMed/NCBI
35	Li M, Jin C, Xu M, Zhou L, Li D and Yin Y: Bifunctional enzyme ATIC promotes propagation of hepatocellular carcinoma by regulating AMPK-mTOR-S6 K1 signaling. Cell Commun Signal. 15:522017. View Article : Google Scholar
36	Che L, Pilo MG, Cigliano A, Latte G, Simile MM, Ribback S, Dombrowski F, Evert M, Chen X and Calvisi DF: Oncogene dependent requirement of fatty acid synthase in hepatocellular carcinoma. Cell Cycle. 16:499–507. 2017. View Article : Google Scholar : PubMed/NCBI
37	Chen YR, Ouyang SS, Chen YL, Li P, Xu HW and Zhu SL: BRD4/8/9 are prognostic biomarkers and associated with immune infiltrates in hepatocellular carcinoma. Aging (Albany NY). 12:17541–17567. 2020. View Article : Google Scholar
38	Kudo M, Kitano M, Sakurai T and Nishida N: general rules for the clinical and pathological study of primary liver cancer, nationwide follow-up survey and clinical practice guidelines: The outstanding achievements of the liver cancer study group of Japan. Dig Dis. 33:765–770. 2015. View Article : Google Scholar : PubMed/NCBI
39	Korean Liver Cancer Study Group (KLCSG); National Cancer Center, Korea (NCC): 2014.Korean liver cancer study group-national cancer center Korea practice guideline for the management of hepatocellular carcinoma. Korean J Radiol. 16:465–522. 2015.