Distribution of variants of the TNFA gene regulatory sites among Russian women of Caucasian origin with leiomyoma

The importance of cytokines, including TNF, in development of uterine fibroids (UF) or leiomyomas (LM) is well proven. The number of polymorphic sites in promoter region of TNF gene is established, and their relationship with the gene expression level has been shown, thus potentially affecting the evolution of the disease. The aim of our research was to analyze the distribution of TNF-238, TNF- 308 and TNF-863 structural variants at the regulatory region of the TNFA gene in a representative group of Caucasian patients with UF compared with a matched group of healthy women followed by assessment of personalized prognostic significance of suggested differences. 180 patients diagnosed with uterine fibroids and 98 practically healthy women were examined. Genotyping of TNF gene polymorphisms (-863 C/A, TNF- 308 G/A, TNF-238 G/A, IL1B-31 T/C, IL4-590 T/C, IL6-174 C/G, IL8-251 T/A, IL10-592 A/G, IL10-1082 A/G and IL17A-197 G/A) was performed by RT-PCR using the SYBR Green intercalating dye. Statistical processing was carried out using multifunctional programs – IBM SPSS Statistics 23, SNPStats. We did not reveal significant differences between the compared groups when analysing distribution of TNF allelic variants and their combinations in genotypes. When comparing the combined SNP variants of the TNF gene at the studied nucleotide positions with the polymorphisms of other genes encoding proinflammatory cytokines, an increased frequency of TNF-238 GG:TNF-308 GG:IL17A-197 AA complex was found in the group of patients with uterine fibroids. The diagnostic specificity of this index was 100%, and the predictive value of this quotient reached 13.3, thus implying a 99.9% probability of a correct prediction. At the same time, the combination of TNF-863 CC genotype with IL-6-174 GG and IL-17-197 GG was significantly reduced in the patients. The frequencies of the anti-inflammatory cytokine genes IL4-590 T/C, IL10-592 A/G and IL10-1082 A/G analyzed n our study did not differ in the both compared groups and they were not considered either predisposing or protective for the disease. The magnitude of the revealed differences in occurrence of these allelic combinations reaches a significant level, thus assuming these traits as potential genetic factors for predicting a predisposal or resistance of women with a certain genotype to development of uterine fibroids, being prognostic criteria for early detection of this disorder.

1. Adamyan L.V., Kuznetsova M.V., Tonoyan N.M., Shapovalenko R.A., Pivazyan L.G., Trofimov D.Yu. Genetic aspects of uterine fibroids: a modern view of the problem. Problemy reproduktsii = Russian Journal of Human Reproduction, 2023, Vol. 29, no. 4‑2, pp. 29‑39. (In Russ.)

2. Konenkov V.I., Prokofiev V.F., Shevchenko A.V., Koroleva E.G., Timofeeva Yu.S., Aidagulova S.V., Marinkin I.O. Characteristics of the cytokine network fragment (TNF–IL-1–IL-4–IL-6–IL-8–IL-10– VEGF) in the peripheral blood of healthy women and patients with leiomyoma. Immunologiya = Immunologiya, 2024, Vol. 45, no. 3, pp. 312–320. (In Russ.)

3. Koroleva Е.G., Konenkov V.I., Shevchenko A.V., Prokofiev V.F., Orlov N.В., Timofeeva Yu.S., Aidagulova S.V., Marinkin I.О. Association of polymorphic variants of cytokines genes, endothelial growth factor and matrix metalloproteinases with the development of uterine fibroids among russian women. Sibirskiy nauchnyy meditsinskiy zhurnal = Siberian Scientific Medical Journal, 2024, Vol. 44, no. 2, pp. 113-122. (In Russ.)

4. Prilepskaya V.N., Serov V.N., Sukhoi G.T. Uterine fibroids. In: Gynecology. Pharmacotherapy without errors. A guide for doctors. Moscow: E-Noto, 2020. pp. 297-305.

5. Sogoyan N.S., Kuznetsova M.V., Donnikov A.E., Mishina N.D., Mikhailovskaya G.V., Shubina E.S., Zelensky D.V., Mullabaeva S.M., Adamyan L.V. Familial predisposition to uterine leiomyoma: searching for genetic factors that increase the risk of leyomyoma development. Problemy reproduktsii = Russian Journal of Human Reproduction, 2020, Vol. 26, no. 5, pp. 51‑57. (In Russ.)

6. Chiricozzi A., Guttman-Yassky E., Suárez-Fariñas M., Nograles K.E., Tian S., Cardinale I., Chimenti S., Krueger J.G. Integrative responses to IL-17 and TNF-α in human keratinocytes account for key inflammatory pathogenic circuits in psoriasis. J. Invest. Dermatol., 2011, Vol. 131, no. 3, pp. 677-687.

7. Ciebiera M., Włodarczyk M., Wrzosek M., Wojtyła C., Błażej M., Nowicka G., Łukaszuk K., Jakiel G. TNF-α serum levels are elevated in women with clinically symptomatic uterine fibroids. Int. J. Immunopathol. Pharmacol., 2018, Vol. 32, 2058738418779461. doi: 10.1177/2058738418779461.

8. Elahi M.M., Asotra K., Matata B.M., Mastana S.S. Tumor necrosis factor alpha −308 gene locus promoter polymorphism: an analysis of association with health and disease. BBA Mol. Basis Dis., 2009, Vol. 1792, no. 3, pp. 163-172.

9. El-Tahan R.R., Ghoneim A.M., El-Mashad N. TNF-α gene polymorphisms and expression. Springerplus, 2016, Vol. 5, no. 1, 1508. doi: 10.1186/s40064-016-3197-y.

10. Hajeer A.H., Hutchinson I.V. Influence of TNFα gene polymorphisms on TNFα production and disease. Hum. Immunol., 2001, Vol. 62, no. 11, pp. 1191-1199.

11. Islam M.S., Protic O., Giannubilo S.R., Toti P., Tranquill, A.L., Petraglia F., Castellucci M., Ciarmela P. Uterine leiomyoma: Available medical treatments and new possible therapeutic options. J. Clin. Endocrinol. Metab., 2013, Vol. 98, no. 3, pp. 921-934.

12. Islam M.S., Ciavattini A., Petraglia F., Castellucci M., Ciarmela P. Extracellular matrix in uterine leiomyoma pathogenesis: a potential target for future therapeutics. Hum. Reprod. Update, 2018, Vol. 24, no.1, pp. 59-85.

13. Krsteski J., Jurgec S., Pakiž M., But I., Potočnik U. Polymorphism of the IL13 gene may be associated with Uterine leiomyomas in Slovenian women. Balk. J. Med. Genet., 2017, Vol. 19, no. 2, pp. 51-60.

14. Kubik-Huch R.A., Weston M., Nougaret S., Leonhardt H., Thomassin-Naggara I., Horta M., Cunha T.M., Maciel C., Rockall A., Forstner R. European Society of Urogenital Radiology (ESUR) Guidelines: MR Imaging of Leiomyomas. Eur. Radiol., 2018, Vol. 28, no. 8, pp. 3125-3137.

15. Medikare V., Altaf A., Ananthapur V., Deendayal M., Nallari P. Susceptibility Risk Alleles of -238G/A, -308G/A and -1031T/C Promoter Polymorphisms of TNF-α Gene to Uterine Leiomyomas. Recent Adv. DNA Gene Seq., 2015, Vol. 9, no. 1, pp. 65-71.

16. Mekinian A., Tamouza R., Pavy S., Gestermann N., Ittah M., Mariette X., Miceli-Richard C. Functional study of TNF-α promoter polymorphisms: literature review and meta-analysis. Eur. Cytokine Netw., 2011, Vol. 22, no. 2, pp. 88-102.

17. Meylan F., Siegel R.M. TNF superfamily cytokines in the promotion of Th9 differentiation and immunopathology. Semin. Immunopathol., 2017, Vol. 39, pp. 21-28.

18. Orciani M., Caffarini M., Biagini A., Lucarini G., Delli Carpini G., Berretta A., Di Primio R., Ciavattini A. Chronic inflammation may enhance leiomyoma development by the involvement of progenitor cells. Stem Cells Int., 2018, Vol. 2018, 1716246. doi: 10.1155/2018/1716246.

19. Pietrowski D., Thewes R., Sator M., Denschlag D., Keck C., Tempfer C. Uterine leiomyoma is associated with a polymorphism in the interleukin 1-beta gene. Am. J. Reprod. Immunol., 2009, Vol. 62, no. 2, pp. 112-117.

20. Shevchenko A.V., Prokofiev V.F., Konenkov V.I., Timopheeva Yu.S., Koroleva E.G., Marinkin I.O., Aidagulova S.A. Features of Matrix Metalloproteinases MMP2, MMP3, and MMP9 of Regulatory-Region Polymorphism in Patients with Uterine Fibroids. Cell Tissue Biol., 2023, Vol. 17, no. 6, pp. 699-705.

21. Tsukamoto K., Ohta N., Shirai Y., Emi M. A highly polymorphic CA repeat marker at the human tumor necrosis factor alpha (TNFAα) locus. J. Hum. Genet., 1998, Vol. 43, no. 4, pp. 278-279.

22. Upadhyay S., Dubey P.K. Gene variants polymorphisms and uterine leiomyoma: an updated review. Front. Genet., 2024, Vol. 15, 1330807. doi: 10.3389/fgene.2024.1330807.