Detection of Novel Autoantibodies to Nucleolin's RNA-binding Domains as a Serum Tumor Biomarker Through ELISA
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Abstract
Expression and location of nucleolin are often abnormal in malignancies, which may result in the production of autoantibodies. Despite this, the identification of such autoantibodies may be essential for the early diagnosis and prognosis of cancers.
In this investigation, the recombinant nucleolin protein was generated using an Escherichia coli expression system and was used an indirect enzyme-linked immunosorbent assay to detect anti-nucleolin autoantibodies in cancer patients' sera.
Lung cancer patients' autoantibodies displayed the highest seroreactivity with the recombinant protein, with area under the curve of 0.948 and sensitivity and specificity of 85% and 96.67%, respectively (accuracy=92%). Anti-nucleolin autoantibodies were linked with lung tumor size (r=0.793), tumor, node, metastasis staging (r=0.643), and proliferation (r=0.744).
These autoantibodies distinguished patients with early-stage lung cancer from healthy controls. Since anti-nucleolin autoantibodies are strongly linked to tumor size, clinical staging, and growth, they can be used to measure how well a treatment is working.
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17. Ezzatifar F RA, Valadan R, Assgarian-omran H, M J-T. Tandem in silico analysis and experimental strategy for designing a novel recombinant RNA binding domain of nucleolin protein for cancer immunotherapy (Under review). Biotechnol Appl Biochem. 2022;15(5):3-9.
18. Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: a laboratory manual: Cold spring harbor laboratory press; 1989.
19. Dai L, Li J, Tsay JJ, Yie TA, Munger JS, Pass H, et al. Identification of autoantibodies to ECH1 and HNRNPA2B1 as potential biomarkers in the early detection of lung cancer. Oncoimmunology. 2017;6(5):e1310359.
20. Katchman BA, Chowell D, Wallstrom G, Vitonis AF, LaBaer J, Cramer DW, et al. Autoantibody biomarkers for the detection of serous ovarian cancer. Gynecol Oncol. 2017;146(1):129-36.
21. Zayakin P, Ancāns G, Siliņa K, Meistere I, Kalniņa Z, Andrejeva D, et al. Tumor-associated autoantibody signature for the early detection of gastric cancer. Int J Cancer. 2013;132(1):137-47.
22. Mattioni M, Soddu S, Prodosmo A, Visca P, Conti S, Alessandrini G, et al. Prognostic role of serum p53 antibodies in lung cancer. BMC Cancer. 2015;15(1):148.
23. Koike Y, Muroi E, Yoshizaki A, Ogawa F, Yanaba K, Takenaka M, et al. Autoantibody against survivin in patients with systemic sclerosis. J Rheumatol. 2010;37(9):1864-70.
24. Minota S, Jarjour WN, Suzuki N, Nojima Y, Roubey RA, Mimura T, et al. Autoantibodies to nucleolin in systemic lupus erythematosus and other diseases. J. Immunol. 1991;146(7):2249-52.
25. Liu J, Yin L, Dong H, Xu E, Zhang L, Qiao Y, et al. Decreased serum levels of nucleolin protein fragment, as analyzed by bead-based proteomic technology, in multiple sclerosis patients compared to controls. J Neuroimmunol. 2012;250(1-2):71-6.
26. Figueiredo J, Miranda A, Lopes-Nunes J, Carvalho J, Alexandre D, Valente S, et al. Targeting nucleolin by RNA G-quadruplex-forming motif. Biochem. Pharmacol. 2021;189:114418.
2. Society A. Cancer treatment & survivorship facts & figures 2019–2021. ACS Atlanta; 2019.
3. Dai L, Lei N, Liu M, Zhang J-Y. Autoantibodies to tumor-associated antigens as biomarkers in human hepatocellular carcinoma (HCC). Exp Hematol Oncol. 2013;2(1):1-7.
4. Tan EM, Zhang J. Autoantibodies to tumor‐associated antigens: reporters from the immune system. Immunol Rev. 2008;222(1):328-40.
5. Qiu J, Choi G, Li L, Wang H, Pitteri SJ, Pereira-Faca SR, et al. Occurrence of autoantibodies to annexin I, 14-3-3 theta and LAMR1 in prediagnostic lung cancer sera. J Clin Oncol. 2008;26(31):5060.
6. Anderson KS, LaBaer J. The sentinel within: exploiting the immune system for cancer biomarkers. J Proteome Res. 2005;4(4):1123-33.
7. Lu H, Goodell V, Disis ML. Humoral immunity directed against tumor-associated antigens as potential biomarkers for the early diagnosis of cancer. J Proteome Res. 2008;7(4):1388-94.
8. Miniard AC, Middleton LM, Budiman ME, Gerber CA, Driscoll DM. Nucleolin binds to a subset of selenoprotein mRNAs and regulates their expression. Nucleic Acids Res. 2010;38(14):4807-20.
9. Chen Z, Xu X. Roles of nucleolin: Focus on cancer and anti-cancer therapy. Saudi Med J 2016;37(12):1312.
10. Tajrishi MM, Tuteja R, Tuteja N. Nucleolin: The most abundant multifunctional phosphoprotein of nucleolus. Commun Integr Biol. 2011;4(3):267-75.
11. Hovanessian AG, Soundaramourty C, Khoury DE, Nondier I, Svab J, Krust B. Surface expressed nucleolin is constantly induced in tumor cells to mediate calcium-dependent ligand internalization. PloS one. 2010;5(12):e15787.
12. Fernandes E, Freitas R, Ferreira D, Soares J, Azevedo R, Gaiteiro C, et al. Nucleolin-Sle A glycoforms as E-selectin ligands and potentially targetable biomarkers at the cell surface of gastric cancer cells. Cancers. 2020;12(4):861.
13. Qiu W, Wang G, Sun X, Ye J, Wei F, Shi X, et al. The involvement of cell surface nucleolin in the initiation of CCR6 signaling in human hepatocellular carcinoma. Med. Oncol. 2015;32(3):1-8.
14. Saadi H, Seillier M, Sandi MJ, Peuget S, Kellenberger C, Gravis G, et al. Development of an ELISA detecting Tumor Protein 53-Induced Nuclear Protein 1 in serum of prostate cancer patients. Results Immunol. 2013;3:51-6.
15. ALabama TUo. The University of ALabama at Birmingham CANcer data analysis Portal Birmingham: The University of Alabama 2022 [An integrated data-mining platform to harness the potential of cancer transcriptome]. Available from: http://ualcan.path.uab.edu.
16. Tang Z, Kang B, Li C, Chen T, Zhang Z. GEPIA2: an enhanced web server for large-scale expression profiling and interactive analysis. Nucleic acids research. 2019;47(W1):W556-w60.
17. Ezzatifar F RA, Valadan R, Assgarian-omran H, M J-T. Tandem in silico analysis and experimental strategy for designing a novel recombinant RNA binding domain of nucleolin protein for cancer immunotherapy (Under review). Biotechnol Appl Biochem. 2022;15(5):3-9.
18. Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: a laboratory manual: Cold spring harbor laboratory press; 1989.
19. Dai L, Li J, Tsay JJ, Yie TA, Munger JS, Pass H, et al. Identification of autoantibodies to ECH1 and HNRNPA2B1 as potential biomarkers in the early detection of lung cancer. Oncoimmunology. 2017;6(5):e1310359.
20. Katchman BA, Chowell D, Wallstrom G, Vitonis AF, LaBaer J, Cramer DW, et al. Autoantibody biomarkers for the detection of serous ovarian cancer. Gynecol Oncol. 2017;146(1):129-36.
21. Zayakin P, Ancāns G, Siliņa K, Meistere I, Kalniņa Z, Andrejeva D, et al. Tumor-associated autoantibody signature for the early detection of gastric cancer. Int J Cancer. 2013;132(1):137-47.
22. Mattioni M, Soddu S, Prodosmo A, Visca P, Conti S, Alessandrini G, et al. Prognostic role of serum p53 antibodies in lung cancer. BMC Cancer. 2015;15(1):148.
23. Koike Y, Muroi E, Yoshizaki A, Ogawa F, Yanaba K, Takenaka M, et al. Autoantibody against survivin in patients with systemic sclerosis. J Rheumatol. 2010;37(9):1864-70.
24. Minota S, Jarjour WN, Suzuki N, Nojima Y, Roubey RA, Mimura T, et al. Autoantibodies to nucleolin in systemic lupus erythematosus and other diseases. J. Immunol. 1991;146(7):2249-52.
25. Liu J, Yin L, Dong H, Xu E, Zhang L, Qiao Y, et al. Decreased serum levels of nucleolin protein fragment, as analyzed by bead-based proteomic technology, in multiple sclerosis patients compared to controls. J Neuroimmunol. 2012;250(1-2):71-6.
26. Figueiredo J, Miranda A, Lopes-Nunes J, Carvalho J, Alexandre D, Valente S, et al. Targeting nucleolin by RNA G-quadruplex-forming motif. Biochem. Pharmacol. 2021;189:114418.
| Files | ||
| Issue | Vol 21 No 6 (2022) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijaai.v21i6.11520 | |
| Keywords | ||
| Autoantibody Biomarker Enzyme-linked immunosorbent assay Neoplasms Nucleolin | ||
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How to Cite
1.
Ezzatifar F, Rafiei A, Valadan R, Asgarian-Omran H, Jeddi-Tehrani M. Detection of Novel Autoantibodies to Nucleolin’s RNA-binding Domains as a Serum Tumor Biomarker Through ELISA. Iran J Allergy Asthma Immunol. 2022;21(6):616-625.
