Current Medicinal ChemistryCurrent Medicinal ChemistryCurrent Medicinal Chemistry0929-86731875-533XBentham Science64516710.2174/0929867330666230503143133Anti-Infectives and Infectious DiseasesResearch ArticleGeneration of Autoantibodies in Metal-catalyzed Oxidatively Damaged DNA in Various Cancer SubjectsAhmadSaheeminfo@benthamscience.netAlshammariQuraininfo@benthamscience.netRafiZeeshaninfo@benthamscience.netRehmanShahnawazinfo@benthamscience.netKhanMohd.info@benthamscience.netFaisalMohd.info@benthamscience.netAlatarAbdulrahmaninfo@benthamscience.netDepartment of Medical Laboratory Sciences, College of Applied Medical Sciences, University of HailDiagnostic Radiology Department, College of Applied Medical Sciences,, University of HailDepartment of Biosciences, Integral UniversityDepartment of Biochemistry, S.S. Faculty of Science, Mohammad Ali Jauhar UniversityDepartment of Biotechnology, School of Applied & Life Science (SALS), Uttaranchal UniversityDepartment of Botany & Microbiology, College of Science,, King Saud University0102202431564064807012025Copyright ©; 2024, Bentham Science Publishers2024Bentham Science Publishershttps://rjraap.com/0929-8673/article/view/645167

Background:Free radicals exist as unstable and highly reactive substances, occurring both in and outside the body. Free radicals are labeled as electron-hungry molecules formed from metabolism and endogenous burning of oxygen. They are transported in cells, upsetting the arrangement of molecules and instigating cellular injury. Hydroxyl radical (•OH) is one of the highly reactive free radicals, which damages the biomolecules in its close vicinity.

Methods:In the present study, DNA was modified by the hydroxyl radical generated via the Fenton reaction. The •OH-oxidized/-modified DNA (Ox-DNA) was characterized by UV-visible and fluorescence spectroscopy. Thermal denaturation was performed to reveal the susceptibility of modified DNA toward heat. The role of Ox-DNA was also established in probing the presence of autoantibodies against Ox-DNA in the sera of cancer patients by direct binding ELISA. The specificity of autoantibodies was also checked by inhibition ELISA.

Results:In biophysical characterization, an increase in hyperchromicity and relative reduction of fluorescence intensity for Ox-DNA was reported compared to the native DNA analog. A thermal denaturation study revealed that Ox-DNA was highly susceptible to heat in comparison to the native conformers. The direct binding ELISA showed the prevalence of autoantibodies from cancer patient sera separated for immunoassay detection against the Ox-DNA. The generated autoantibodies against the Ox-DNA were detected as highly specific against bladder, head, neck, and lung cancer, which was further confirmed by the inhibition ELISA for the serum and IgG antibodies.

Conclusion:The generated neoepitopes on DNA molecules are recognized as nonself by the immune system, which leads to the formation of autoantibodies in cancer patients. Therefore, our study confirmed that oxidative stress plays a role in the structural perturbation of DNA and makes it immunogenic.

Hydroxyl radical (•OH)fenton reactionneoepitopesautoantibodiescancerdamaged DNA.Halliwell, B. Protection against oxidants in biological systems. The superoxide theory of oxygen toxicity. Biol. Med., 1989, 86-123.Zeng, W.; Zhang, H.; Yuan, X.; Chen, T.; Pei, Z.; Ji, X. Two-dimensional nanomaterial-based catalytic medicine: Theories, advanced catalyst and system design. Adv. Drug Deliv. Rev., 2022, 184, 114241. doi: 10.1016/j.addr.2022.114241 PMID: 35367308Rafi, Z.; Alouffi, S.; Khan, M.S.; Ahmad, S. 2′-Deoxyribose mediated glycation leads to alterations in BSA structure via generation of carbonyl species. Curr. Protein Pept. Sci., 2020, 21(9), 924-935. doi: 10.2174/1389203721666200213104446 PMID: 32053073Ahmad, S.; Rehman, S.; Alsulimani, A.; Al-shaghdali, K.; Khan, M. Y. Non-enzymatic glycosylation of isolated human Immunoglobulin-G by D-Ribose. Cell Biochem. Funct., 2022, 40(5), 526-534. doi: 10.1002/cbf.3722 PMID: 35707967Rehman, S.; Mohammad, A.; Zeeshan, R. Effect of non-enzymatic glycosylation in the epigenetics of cancer. Semin. Cancer Biol., 2020, 83, 543-555. doi: 10.1016/j.semcancer.2020.11.019 PMID: 33276090Ahmad, S.; Khan, H.; Siddiqui, Z.; Khan, M.Y.; Rehman, S.; Shahab, U.; Godovikova, T.; Silnikov, V.; Moinuddin AGEs, RAGEs and s-RAGE; friend or foe for cancer. Semin. Cancer Biol., 2018, 49, 44-55. doi: 10.1016/j.semcancer.2017.07.001 PMID: 28712719Ji, X.; Ge, L.; Liu, C.; Tang, Z.; Xiao, Y.; Chen, W.; Lei, Z.; Gao, W.; Blake, S.; De, D.; Shi, B.; Zeng, X.; Kong, N.; Zhang, X.; Tao, W. Capturing functional two-dimensional nanosheets from sandwich-structure vermiculite for cancer theranostics. Nat. Commun., 2021, 12(1), 1124. doi: 10.1038/s41467-021-21436-5 PMID: 33602928Bartsch, H. DNA adducts in human carcinogenesis: Etiological relevance and structure-activity relationship. Mutat. Res. Rev. Genet. Toxicol., 1996, 340(2-3), 67-79. doi: 10.1016/S0165-1110(96)90040-8 PMID: 8692183Ahmad, S.; Firoz, A.; Uzma, S. Do all roads lead to the Rome? The glycation perspective! Semin. Cancer Biol., 2017, 49, 9-19. doi: 10.1016/j.semcancer.2017.10.012 PMID: 29113952Ahmad, S. Physicochemical characterization of in vitro ldl glycation and its inhibition by ellagic acid (EA): An in vivo approach to inhibit diabetes in experimental animals. Biomed Res Int, 2022, 2022, 5583298. doi: 10.1155/2022/5583298 PMID: 35097119Kong, N.; Zhang, H.; Feng, C. Arsenene-mediated multiple independently targeted reactive oxygen species burst for cancer therapy. Nat. Commun, 2021, 12(1), 1-8. doi: 10.1038/s41467-021-24961-5Lobo, V.; Patil, A.; Phatak, A.; Chandra, N. Free radicals, antioxidants and functional foods: Impact on human health. Pharmacogn. Rev., 2010, 4(8), 118-126. doi: 10.4103/0973-7847.70902 PMID: 22228951Ahmad, S.; Khan, H.; Shahab, U.; Rehman, S.; Rafi, Z.; Khan, M.Y.; Ansari, A.; Siddiqui, Z.; Ashraf, J.M.; Abdullah, S.M.; Habib, S.; Uddin, M. Protein oxidation an overview of metabolism of sulphur containing amino acid cysteine. Front. Biosci., 2017, 9(1), 71-87. doi: 10.2741/s474 PMID: 27814576Moinuddin; Dixit, K.; Ahmad, S.; Shahab, U.; Habib, S.; Naim, M.; Alam, K.; Ali, A. Human DNA damage by the synergistic action of 4-aminobiphenyl and nitric oxide: An immunochemical study. Environ. Toxicol., 2014, 29(5), 568-576. doi: 10.1002/tox.21782 PMID: 22610904Khan, M.Y.; Alouffi, S.; Ahmad, S. Immunochemical studies on native and glycated LDL – An approach to uncover the structural perturbations. Int. J. Biol. Macromol., 2018, 115, 287-299. doi: 10.1016/j.ijbiomac.2018.04.016 PMID: 29634967Rehman, S.; Alouffi, S.; Faisal, M.; Qahtan, A.A.; Alatar, A.A.; Ahmad, S. Methylglyoxal mediated glycation leads to neo-epitopes generation in fibrinogen: Role in the induction of adaptive immune response. Int. J. Biol. Macromol., 2021, 175, 535-543. doi: 10.1016/j.ijbiomac.2021.01.197 PMID: 33529635Shahab, U.; Ahmad, S.; Moinuddin; Dixit, K.; Habib, S.; Alam, K.; Ali, A. Hydroxyl radical modification of collagen type II increases its arthritogenicity and immunogenicity. PLoS One, 2012, 7(2), e31199. doi: 10.1371/journal.pone.0031199 PMID: 22319617Ahmad, S.; Moinuddin; Dixit, K.; Shahab, U.; Alam, K.; Ali, A. Genotoxicity and immunogenicity of DNA-advanced glycation end products formed by methylglyoxal and lysine in presence of Cu2+. Biochem. Biophys. Res. Commun., 2011, 407(3), 568-574. doi: 10.1016/j.bbrc.2011.03.064 PMID: 21420380Shahab, U.; Moinuddin; Ahmad, S.; Dixit, K.; Abidi, S.M.A.; Alam, K.; Ali, A. Acquired immunogenicity of human DNA damaged by N-hydroxy-N-acetyl-4-aminobiphenyl. IUBMB Life, 2012, 64(4), 340-345. doi: 10.1002/iub.1008 PMID: 22378795Siddiqui, Z.; Ishtikhar, M.; Moinuddin; Ahmad, S. d-Ribose induced glycoxidative insult to hemoglobin protein: An approach to spot its structural perturbations. Int. J. Biol. Macromol., 2018, 112, 134-147. doi: 10.1016/j.ijbiomac.2018.01.161 PMID: 29378270Mustafa, I.; Ahmad, S.; Dixit, K.; Moinuddin; Ahmad, J.; Ali, A. Glycated human DNA is a preferred antigen for anti-DNA antibodies in diabetic patients. Diabetes Res. Clin. Pract., 2012, 95(1), 98-104. doi: 10.1016/j.diabres.2011.09.018 PMID: 22001283Shahab, U.; Moinuddin; Ahmad, S.; Dixit, K.; Habib, S.; Alam, K.; Ali, A. Genotoxic effect of N-hydroxy-4-acetylaminobiphenyl on human DNA: implications in bladder cancer. PLoS One, 2013, 8(1), e53205. doi: 10.1371/journal.pone.0053205 PMID: 23382838Ahmad, S.; Moinuddin; Shahab, U.; Habib, S.; Salman Khan, M.; Alam, K.; Ali, A. Glycoxidative damage to human DNA: Neo-antigenic epitopes on DNA molecule could be a possible reason for autoimmune response in type 1 diabetes. Glycobiology, 2014, 24(3), 281-291. doi: 10.1093/glycob/cwt109 PMID: 24347633Ashraf, J.M.; Ahmad, S.; Rabbani, G.; Hasan, Q.; Jan, A.T.; Lee, E.J.; Khan, R.H.; Alam, K.; Choi, I. 3-Deoxyglucosone: A potential glycating agent accountable for structural alteration in H3 histone protein through generation of different AGEs. PLoS One, 2015, 10(2), e0116804. doi: 10.1371/journal.pone.0116804 PMID: 25689368Faisal, M.; Shahab, U.; Alatar, A.A.; Ahmad, S. Preferential recognition of auto-antibodies against 4- hydroxynonenal modified DNA in the cancer patients. J. Clin. Lab. Anal., 2017, 31(6), e22130. doi: 10.1002/jcla.22130 PMID: 28105689Akhter, F.; Khan, M.S.; Singh, S.; Ahmad, S. An immunohistochemical analysis to validate the rationale behind the enhanced immunogenicity of D-ribosylated low density lipo-protein. PLoS One, 2014, 9(11), e113144. doi: 10.1371/journal.pone.0113144 PMID: 25393017