Association of HLA-B27 Gene and Rheumatoid Arthritis: Analysis of Potential Role as a Predictive Biomarker

Karzan Ghafur Khidhir; Baban Ahmad; Dana Khdr Sabir

doi:10.24017/science.2023.2.3

Authors

Karzan Ghafur Khidhir Department of Biology, College of Science, University of Sulaimani, Sulaymaniyah, Iraq https://orcid.org/0000-0003-3803-2588
Baban Ahmad Genetics laboratory, Harem Hospital, Sulaymaniyah, Iraq
Dana Khdr Sabir Department of Medical Laboratory Science, College of Medicals and Applied Sciences, Charmo University, Chamchamal, Iraq. https://orcid.org/0000-0001-6197-7737

Abstract

Rheumatoid arthritis (RA) is a predominant inflammatory arthritis in human. The function of HLA-B27 gene in other types of arthritis has been studied, however, its function in RA is unclear. This study investigated the relative expression of HLA-B27 gene in RA patients compared to normal control and assesses its suitability as a biomarker for early detection of RA. Bioinformatics analysis was used to determine the profile of HLA-B27 gene in different human tumors to demonstrate the possible involvement of HLA-B27 in both RA and cancer; and also, to discover its functional association with other human genes. Samples of human blood from RA patients and healthy individuals were collected, and RNA extraction, cDNA synthesis and qPCR were carried out to detect relative expression of HLA-B27. ROC analysis was undertaken to investigate HLA-B27 diagnostic performance; GENT2 platform was used to compare HLA-B27 expression levels in different human tumors, and gene-gene interaction network was generated using GeneMANIA to identify correlation of HLA-B27 with other human genes. The qPCR analysis demonstrated an increase in the HLA-B27 expression by 1.65 fold in RA compared to normal control. ROC analysis indicated that HLA-B27 expression could efficiently differentiate RA from normal, supporting its potential use as diagnostic molecular biomarkers. The GENT2 revealed that HLA-B27 expression levels vary across different tumor types, most notably in heart tissue. The gene-gene interaction network revealed that KIR3DL1, KIR3DS1, LILRB1, B2M and LILRA1 were the leading genes showing the highest correlations with the HLA-B27. Our results indicate that HLA-B27 gene is involved in the RA pathogenesis and it can be used as a molecular biomarker for the diagnosis of RA. Our findings could lead to the discovery of novel diagnostic, preventive and therapeutic strategies.

Keywords:

Rheumatoid arthritis, Molecular Biomarker, HLA-B27, qPCR, ROC

References

G. S. Firestein, "Evolving concepts of rheumatoid arthritis," Nature, vol. 423, pp. 356-361, 2003. doi: 10.1038/nature01661. DOI: https://doi.org/10.1038/nature01661

S. E. Gabriel, "The epidemiology of rheumatoid arthritis," Rheum Dis. Clin North Am., vol. 27, pp. 269-281, 2001. doi: 10.1016/s0889-857x(05)70201-5. DOI: https://doi.org/10.1016/S0889-857X(05)70201-5

B. M. Köhler, J. Günther, D. Kaudewitz, and H.-M. Lorenz, "Current therapeutic options in the treatment of rheumatoid arthritis," J. Clin. Med., vol. 8, p. 938, 2019. doi: h10.3390/jcm807-0938. DOI: https://doi.org/10.3390/jcm8070938

H. U. Scherer, T. Häupl, and G. R. Burmester, "The etiology of rheumatoid arthritis," J. Autoimmun., vol. 110, p. 102400, 2020. doi: 10.1016/j.jaut.2019.102400. DOI: https://doi.org/10.1016/j.jaut.2019.102400

T. C. Messemaker, T. W. Huizinga, and F. Kurreeman, "Immunogenetics of rheumatoid arthritis: understanding func-tional implications," J.Autoimmun., vol. 64, pp. 74-81, 2015,doi: 10.1016/j.jaut.2015.07.007. DOI: https://doi.org/10.1016/j.jaut.2015.07.007

C. M. Weyand and J. J. Goronzy, "The immunology of rheumatoid arthritis," Nat. Immunol, vol. 22, pp. 10-18, 202. doi: 10.1038/s41590-020-00816-x. DOI: https://doi.org/10.1038/s41590-020-00816-x

E. G. Favalli, F. Ingegnoli, O. De Lucia, G. Cincinelli, R. Cimaz, and R. Caporali, "COVID-19 infection and rheumatoid arthritis: Faraway, so close!," Autoimmun. Rev., vol. 19, p. 102523, 2020. doi: 10.1016/j.autrev.2020.102523. DOI: https://doi.org/10.1016/j.autrev.2020.102523

M. J. Peters, I. E. van der Horst-Bruinsma, B. A. Dijkmans, and M. T. Nurmohamed, "Cardiovascular risk profile of pa-tients with spondylarthropathies, particularly ankylosing spondylitis and psoriatic arthritis," Semin. Arthritis Rheum, 2004, pp. 585-592. doi: 10.1016/j.semarthrit-.2004.07.010. DOI: https://doi.org/10.1016/j.semarthrit.2004.07.010

D. Van der Woude, et al., "Quantitative heritability of anti–citrullinated protein antibody–positive and anti–citrullinated protein antibody–negative rheumatoid arthritis," Arthritis Rheum., vol. 60, pp. 916-923, 2009. doi: http://doi.org/10.1002/art.24385. DOI: https://doi.org/10.1002/art.24385

J. Smolen, D. Aletaha, and I. McInnes, "Rheumatoid arthritis. Lancet, vol. 388, no. 10055, pp. 2023–2038, 2016. doi:10.1016/S0140-6736(16)30173-8. DOI: https://doi.org/10.1016/S0140-6736(16)30173-8

Y. Okada, et al., "Genetics of rheumatoid arthritis contributes to biology and drug discovery," Nature, vol. 506, pp. 376-381, 2014. doi: 10.1038/-nature12873.

L. E. Dedmon, "The genetics of rheumatoid arthritis," Rheumatology, vol. 59, pp. 2661-2670, 2020. doi: 10.1093/-rheumatology/keaa232. DOI: https://doi.org/10.1093/rheumatology/keaa232

J. Hammer, et al., "Peptide binding specificity of HLA-DR4 molecules: correlation with rheumatoid arthritis associa-tion," J.Exp Med., vol. 181, pp. 1847-1855, 1995. doi: 10.1084/jem.181.5.1847. DOI: https://doi.org/10.1084/jem.181.5.1847

P. K. Gregersen, J. Silver, and R. J. Winchester, "The shared epitope hypothesis. An approach to understanding the mo-lecular genetics of susceptibility to rheumatoid arthritis," Arthritis Rheum., vol. 30, pp. 1205-1213, 1987. doi: 10.1002/-art.1780301102. DOI: https://doi.org/10.1002/art.1780301102

M. A. Khan, "HLA-B27 and its pathogenic role," J. Clin. Rheumatol., vol. 14, pp. 50-52, 2008. doi: 10.1097/RHU.0b013e3181637-a38. DOI: https://doi.org/10.1097/RHU.0b013e3181637a38

D. Wakefield, D. Clarke, and P. McCluskey, "Recent developments in HLA B27 anterior uveitis," Front. Immunol, vol. 11, p. 608134, 2021. doi: 10.3389/fimmu-.2020.608134. DOI: https://doi.org/10.3389/fimmu.2020.608134

P. Bowness, "HLA-B27," Annu. Rev. Immunol, vol. 33, pp. 29-48, 2015. doi: 10.1146/annurev-immunol-032414-112110. DOI: https://doi.org/10.1146/annurev-immunol-032414-112110

Y. M. Chung, et al., "Prevalence of spondyloarthritis in 504 Chinese patients with HLA‐B27‐associated acute anterior uveitis," Scand. J. Rheumatol., vol. 38, pp. 84-90, 2009. doi: 10.1080/03009740802385423. DOI: https://doi.org/10.1080/03009740802385423

A. M. Braakenburg, H. W. De Valk, J. De Boer, and A. Rothova, "Human Leukocyte Antigen-B27–associated uveitis: Long-term follow-up and gender differences," Am.J. Ophthalmol., vol. 145, pp. 472-479, 2008. doi: 10.1016/j.ajo.2007.11.009. DOI: https://doi.org/10.1016/j.ajo.2007.11.009

M. Adam, A. N. Erkan, D. Arslan, B. Leblebici, L. Özlüoğlu, and M. N. Akman, "High-frequency sensorineural hearing loss in patients with ankylosing spondylitis: is it an extrarticuler feature of disease?," Rheumatol. Int., vol. 28, pp. 413-417, 2008. doi: 10.1007/s00296-007-0458-7. DOI: https://doi.org/10.1007/s00296-007-0458-7

D. Aletaha and J. S. Smolen, "Diagnosis and management of rheumatoid arthritis: a review," JAMA, vol. 320, pp. 1360-1372, 2018. doi: 10.1001/jama.2018.13103. DOI: https://doi.org/10.1001/jama.2018.13103

M. C. Hochberg, R. W. Chang, I. Dwosh, S. Lindsey, T. Pincus, and F. Wolfe, "The American College of Rheumatology 1991 revised criteria for the classification of global functional status in rheumatoid arthritis," Arthritis Rheum., vol. 35, pp. 498-502, 1992. doi: 10.1002/art.1780350502. DOI: https://doi.org/10.1002/art.1780350502

A. Kurata, et al., "Expression level of microRNA-200c is associated with cell morphology in vitro and histological dif-ferentiation through regulation of ZEB1/2 and E-cadherin in gastric carcinoma," Oncol. Rep., vol. 39, pp. 91-100, 2018. doi: 10.3892/or.2017.6093. DOI: https://doi.org/10.3892/or.2017.6093

M. A. Bon, A. van Oeveren-Dybicz, and F. A. van den Bergh, "Genotyping of HLA-B27 by real-time PCR without hy-bridization probes," Clin. Chem., vol. 46, pp. 1000-1002, 2000. doi: 10.1093/-clinchem/46.7.1000. DOI: https://doi.org/10.1093/clinchem/46.7.1000

T. D. Schmittgen and K. J. Livak, "Analyzing real-time PCR data by the comparative C T method," Nat. Protoc, vol. 3, p. 1101, 2008. doi: 10.1038-/nprot.2008.73. DOI: https://doi.org/10.1038/nprot.2008.73

Y. Nätterkvist, "Development of a PCR method to detect HLA-B27 in ankylosing spondylitis," 2012. https://www.diva-portal.org/smash/get/diva2:565006/FULLTEXT01.pdf.

S.-J. Park, B.-H. Yoon, S.-K. Kim, and S.-Y. Kim, "GENT2: an updated gene expression database for normal and tumor tissues," BMC Med.Genomics, vol. 12, pp. 1-8, 2019. doi:10.1186/s12920 -019-0514-7. DOI: https://doi.org/10.1186/s12920-019-0514-7

M. Franz, et al., "GeneMANIA update 2018," Nucleic acids Res, vol. 46, pp. W60-W64, 2018. doi:10.1093/nar/gky311. DOI: https://doi.org/10.1093/nar/gky311

M. D. Ruopp, N. J. Perkins, B. W. Whitcomb, and E. F. Schisterman, "Youden Index and optimal cut‐point estimated from observations affected by a lower limit of detection," Biom. J., vol. 50, pp. 419-430, 2008. doi: 10.1002/bimj.200710415. DOI: https://doi.org/10.1002/bimj.200710415

R. Deshmukh, "Rheumatoid arthritis: pathophysiology, current therapeutic strategies and recent advances in targeted drug delivery system," Mater. Today Commun., vol. 35. 105877, 2023. doi: 10.3390/cells10113017. DOI: https://doi.org/10.1016/j.mtcomm.2023.105877

M. H. Nicknam, et al., "Determination of HLA-B27 subtypes in Iranian patients with ankylosing spondylitis," Iranian J. Allergy Asthma Immunol., vol. 7, no. 1, pp. 19-24, 2008. PMID: 18322308.

E. Diyarbakir, N. Eyerci, M. Melikoglu, A. Topcu, and I. Pirim, "HLA B27 subtype distribution among patients with ankylosing spondylitis in eastern Turkey," Genet. Test Mol. Biomarkers, vol. 16, pp. 456-458, 2012. doi: 10.1089/gtmb.2011-.0183 DOI: https://doi.org/10.1089/gtmb.2011.0183

K. Esalat-Manesh, M. Taghadosi, and A. Arj, "A survey on the frequency of HLA-B27 in patients engaged with seroneg-ative spondyloarthropathies in Kashan, Iran," Zahedan J. Res. Med. Sci., vol. 17, no. 6, pp. e983, 2015. doi: 10.17795/-zjrms983. DOI: https://doi.org/10.17795/zjrms983

H. Yazici, I. Schreuder, S. Yurdakul, and F. Ozbakir, "HLA-B27 in Turkish patients with rheumatoid arthritis," Ann. Rheum. Dis, vol. 46, pp. 718, 1987. doi: 10.1136/ard.46-.9.718 DOI: https://doi.org/10.1136/ard.46.9.718

N. R. Ziade, "HLA B27 antigen in Middle Eastern and Arab countries: systematic review of the strength of association with axial spondyloarthritis and methodological gaps," BMC Musculoskelet. Disord., vol. 18, pp. 1-5, 2017. doi: 10.1186/s12891 -017-1639-5. DOI: https://doi.org/10.1186/s12891-017-1639-5

K. N. Mustafa, M. Hammoudeh, and M. A. Khan, "HLA-B27 prevalence in Arab populations and among patients with ankylosing spondylitis," J. Rheum., vol. 39, pp. 1675-1677, 2012. doi:10.3899/jrheum.120403. DOI: https://doi.org/10.3899/jrheum.120403

A. Yamaguchi, et al., "Association of hla‐b39 with hla‐b27‐negative ankylosing spondylitis and pauciarticular juvenile rheumatoid arthritis in japanese patients," Arthritis Rheum., vol. 38, pp. 1672-1677, 1995. doi: 10.1002/art.-1780381120. DOI: https://doi.org/10.1002/art.1780381120

C. Perricone, F. Ceccarelli, and G. Valesini, "An overview on the genetic of rheumatoid arthritis: a never-ending story," Autoimmun. Rev., vol. 10, pp. 599-608, 2011. doi: 10.3390/life-11060524. DOI: https://doi.org/10.1016/j.autrev.2011.04.021

Y. Kochi, A. Suzuki, and K. Yamamoto, "Genetic basis of rheumatoid arthritis: a current review," BiochemBiophys. Res. Commun., vol. 452, pp. 254-262, 2014. doi: 10.1016/j.bbrc.2014.07.085. DOI: https://doi.org/10.1016/j.bbrc.2014.07.085

S. Sardar, et al., "The oncoprotein TBX3 is controlling severity in experimental arthritis," Arthritis Res. Ther., vol. 21, pp. 1-17, 2019. doi: 10.1186/s13075-018-1797-3. DOI: https://doi.org/10.1186/s13075-018-1797-3

R. M. Plenge, et al., "Replication of putative candidate-gene associations with rheumatoid arthritis in> 4,000 samples from North America and Sweden: association of susceptibility with PTPN22, CTLA4, and PADI4," Am. J. Hum. Genet., vol. 77, pp. 1044-1060, 2005. doi: 10.1086/498651. DOI: https://doi.org/10.1086/498651

L. Padyukov, "Genetics of rheumatoid arthritis," Semin. Immunopathol., vol. 44, no. 1, pp. 47-62, 2022. doi: 10.1007/s00281-022-00912-0. DOI: https://doi.org/10.1007/s00281-022-00912-0

L. Mellemkjær, M. Linet, G. Gridley, M. Frisch, H. Møller, and J. Olsen, "Rheumatoid arthritis and cancer risk," Eur. J. Cancer, vol. 32, no. 10, pp. 1753-1757, 1996. doi: 10.1016/0959-8049(96)-00210-9. DOI: https://doi.org/10.1016/0959-8049(96)00210-9

A. Parikh-Patel, R. H. White, M. Allen, and R. Cress, "Risk of cancer among rheumatoid arthritis patients in California," Cancer Causes Control, vol. 20, pp. 1001-1010, 2009. doi: 10.1007/s10552-009-9298-y. DOI: https://doi.org/10.1007/s10552-009-9298-y

Y. J. Chen, Y. T. Chang, C. B. Wang, and C. Y. Wu, "The risk of cancer in patients with rheumatoid arthritis: a nation-wide cohort study in Taiwan," Arthritis Rheum., vol. 63, no. 2, pp. 352-358, 2011. doi: 10.1002/art.30134. DOI: https://doi.org/10.1002/art.30134

X. Pundole and M. E. Suarez-Almazor, "Cancer and rheumatoid arthritis," Rheum. Dis. Clin. North Am., vol. 46, no. 3, pp. 445-462, 2020, doi:10.1016/j.rdc.-2020.05.003. DOI: https://doi.org/10.1016/j.rdc.2020.05.003

E. Rajaei, H. Haybar, K. Mowla, and Z. D. Zayeri, "Metformin one in a million efficient medicines for rheumatoid ar-thritis complications: Inflammation, osteoblastogenesis, cardiovascular disease, malignancies," Curr. Rheumatol. Rev., vol. 15, no. 2, pp. 116-122, 2019. doi:10.2174/1573397114666180717145745. DOI: https://doi.org/10.2174/1573397114666180717145745

J. Kishikawa, et al., "Characteristics and prognosis of colorectal cancer associated with rheumatic disease," Int. Surg., vol. 100, no. 5, pp. 783-789, 2015. doi: 10.9738/INTSURG-D-14-00154.1. DOI: https://doi.org/10.9738/INTSURG-D-14-00154.1

S. Bergamaschi, et al., "Tumor markers are elevated in patients with rheumatoid arthritis and do not indicate presence of cancer," Int. J. Rheum. Dis., vol. 15, no. 2, pp. 179-182, 2012. doi: 10.1111/j.1756-185X.2011.01671.x. DOI: https://doi.org/10.1111/j.1756-185X.2011.01671.x

T. A. Simon, A. Thompson, K. K. Gandhi, M. C. Hochberg, and S. Suissa, "Incidence of malignancy in adult patients with rheumatoid arthritis: a meta-analysis," Arthritis Res. Ther., vol. 17, pp. 1-10, 2015. doi: 10.1186/s13075-015-0728-9. DOI: https://doi.org/10.1186/s13075-015-0728-9

H.-W. Tseng, L.-Y. Lu, H.-C. Lam, K.-W. Tsai, W.-C. Huang, and Y.-L. Shiue, "The influence of disease-modifying an-ti-rheumatic drugs and corticosteroids on the association between rheumatoid arthritis and skin cancer: a nationwide retrospective case-control study in Taiwan," Clin. Exp. Rheumatol., vol. 36, no. 3, pp. 471-8, 2018. PMID: 29303707.

E. Schaafsma, C. M. Fugle, X. Wang, and C. Cheng, "Pan-cancer association of HLA gene expression with cancer prog-nosis and immunotherapy efficacy," Br. J. Cancer, vol. 125, no. 3, pp. 422-432, 2021. doi: 10.1038/s41416-021-01400-2. DOI: https://doi.org/10.1038/s41416-021-01400-2

N. Aptsiauri and F. Garrido, "The Challenges of HLA Class I Loss in Cancer Immunotherapy: Facts and Hopes," Clin. Cancer Res., vol. 28, no. 23, pp. 5021-5029, 2022. doi: 10.1158/1078-0432.CCR-21-3501. DOI: https://doi.org/10.1158/1078-0432.CCR-21-3501

H. Liang, T. Lu, H. Liu, and L. Tan, "The Relationships between HLA-A and HLA-B Genes and the Genetic Susceptibil-ity” Russ. J. Genet., vol. 57, pp. 1206-1213, 2021. doi: 10.1134/S1022795421100069. DOI: https://doi.org/10.1134/S1022795421100069