{"paper_id":"afb89ced-9295-4013-a596-fde541a1938f","body_text":"Abstract\nBackground\nEndometriosis (EM) is defined as gynecological chronic inflammation and the presence of active foci of uterine mucous membrane outside the uterine cavity. Susceptibility to endometriosis is influenced by single nucleotide polymorphisms (SNPs) in intronic or intergenic regions. This study aimed to analyze the frequencies of genotypes and alleles of LEP G2548A (rs7799039) of the LEP gene and LEPR Q223R (rs1137101) of the LEPR gene to investigate their contributions to the pathogenesis of endometriosis.\nMethods\nPeripheral blood samples of 178 endometriosis patients and 160 healthy donors were isolated from peripheral blood leukocytes. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) techniques were used. A novel Bioinformatics analysis was undertaken to anticipate the potential roles of these polymorphisms.\nResults\nThe LEP rs7799039 AA genotype and mutant allele (A) were frequent in EM women as compared to healthy subjects (OR = 2.35, p = 0.024; OR = 1.35, p = 0.048, respectively). The LEPR rs1137101 mutant GG genotype mutant allele (G) was related to an increased risk of endometriosis (OR=3.22, p = 0.003; OR=1.45, p = 0.022, respectively). The combination of mutant AA and GG genotypes of LEP and LEPR SNPs showed an extremely adverse effect on disease susceptibility and more than ten times more susceptibility to EM. In addition, carriers of the mutant allele for LEP rs7799039 and LEPR rs1137101 polymorphism were more frequent in the moderate/severe (stage III and IV) group of the disease. The LEP G2548A (rs7799039) gene polymorphism may lead to varying allelic expression, as indicated by an in-silico research. Furthermore, bioinformatics analysis indicated that the LEPR Q223R (rs1137101) SNP will significantly alter its secondary structure and physicochemical properties.\nConclusion\nOur research indicates that the LEP G2548A (rs7799039) and LEPR Q223R (rs1137101) polymorphisms may play a role in the pathophysiology of endometriosis. Considering the proven association between leptin signaling and endometriosis, these SNPs may function as potential indicators of disease risk.\nSimilar content being viewed by others\nData Availability\nThe data presented in this study are available on request from the corresponding author.\nReferences\nAcién P, Velasco I. Endometriosis: a disease that remains enigmatic. ISRN Obstet Gynecol. 2013;2013:1–12.\nZondervan KT, Becker CM, Koga K, et al. Endometriosis. Nat Rev Dis Primers. 2018; 4 (1): 9. PubMed Artic.\nMoradi Y, Shams-Beyranvand M, Khateri S, et al. A systematic review on the prevalence of endometriosis in women. Indian J Med Res. 2021;154(3):446–54.\nLaganà AS, Garzon S, Götte M, et al. The pathogenesis of endometriosis: molecular and cell biology insights. Int J Mol Sci. 2019;20(22):5615.\nMatalliotakis M, Zervou MI, Matalliotaki C, et al. The role of gene polymorphisms in endometriosis. Mol Med Rep. 2017;16(5):5881–6.\nPérez-Pérez A, Toro A, Vilariño-García T, et al. Leptin action in normal and pathological pregnancies. J Cell Mol Med. 2018;22(2):716–27.\nKalaitzopoulos DR, Lempesis IG, Samartzis N, et al. Leptin concentrations in endometriosis: a systematic review and meta-analysis. J Reprod Immunol. 2021;146: 103338.\nOh HK, Choi YS, Yang YI, et al. Leptin receptor is induced in endometriosis and leptin stimulates the growth of endometriotic epithelial cells through the JAK2/STAT3 and ERK pathways. Mol Hum Reprod. 2013;19(3):160–8.\nMantzoros CS, Magkos F, Brinkoetter M, et al. Leptin in human physiology and pathophysiology. Am J Physiol Endocrinol Metab. 2011;301(4):E567-84.\nHussein SS, Farhan FS, Ali AI. Serum leptin as a marker for severity of endometriosis. Obstet Gynecol Int. 2020;2020:1–5.\nPandey N, Kriplani A, Yadav RK, et al. Peritoneal fluid leptin levels are increased but adiponectin levels are not changed in infertile patients with pelvic endometriosis. Gynecol Endocrinol. 2010;26(11):843–9.\nTian Z, Wang Y, Zhao Y, et al. Serum and peritoneal fluid leptin levels in endometriosis: a systematic review and meta-analysis. Gynecol Endocrinol. 2021;37(8):689–93.\nMatarese G, Alviggi C, Sanna V, et al. Increased leptin levels in serum and peritoneal fluid of patients with pelvic endometriosis. J Clin Endocrinol Metab. 2000;85(7):2483–7.\nZhao Z, Wu Y, Zhang H, et al. Association of leptin and adiponectin levels with endometriosis: a systematic review and meta-analysis. Gynecol Endocrinol. 2021;37(7):591–9.\nZhu S, Tang Z, Tang Y, et al. Correlation of Q223R and K109R polymorphisms in leptin receptor gene with susceptibility of breast cancer: a systematic review and meta-analysis. J Chin Med Assoc. 2023;86(6):549–56.\nYuan Q, Zhang L, Tian Y, et al. Circulating leptin level, soluble leptin receptor level and their gene polymorphism in patients with systemic lupus erythematosus: a systematic review and meta-analysis. Clin Exp Rheumatol. 2020;38(6):1238–46.\nKhaki-Khatibi F, Shademan B, Gholikhani-Darbroud R, et al. Gene polymorphism of leptin and risk for heart disease, obesity, and high BMI: a systematic review and pooled analysis in adult obese subjects. Horm Mol Biol Clin Investig. 2023;44(1):11–20.\nWu L, Sun D. Leptin receptor gene polymorphism and the risk of cardiovascular disease: a systemic review and meta-analysis. Int J Environ Res Public Health. 2017;14(4):375.\nGhalandari H, Hosseini-Esfahani F, Mirmiran P. The association of polymorphisms in leptin/leptin receptor genes and ghrelin/ghrelin receptor genes with overweight/obesity and the related metabolic disturbances: A review. Int J Endocrinol Metab. 2015;13(3).\nDuarte SFP, Francischetti EA, Genelhu VA, et al. LEPR p.Q223R, β3-AR p.W64R and LEP c.-2548G>A gene variants in obese Brazilian subjects. Genet Mol Res. 2007;6(4):1035–43.\nHoffsted J, Eriksson P, Mottagui-Tabar S, et al. A polymorphism in the leptin promoter region (-2548 G/A) influences gene expression and adipose tissue secretion of leptin. Horm Metab Res. 2002;34(7):355–9.\nYiannakouris N, Yannakoulia M, Melistas L, et al. The Q223R polymorphism of the leptin receptor gene is significantly associated with obesity and predicts a small percentage of body weight and body composition variability. J Clin Endocrinol Metab. 2001;86(9):4434–9.\nChin JR, Heuser CC, Eller AG, et al. Leptin and leptin receptor polymorphisms and recurrent pregnancy loss. J Perinatol. 2013;33(8):589–92.\nAli EMM, Diab T, Elsaid A, et al. Fat mass and obesity-associated (FTO) and leptin receptor (LEPR) gene polymorphisms in Egyptian obese subjects. Arch Physiol Biochem. 2021;127(1):28–36.\nKeppler-Noreuil KM, Parker VER, Darling TN, et al. Somatic overgrowth disorders of the PI3K/AKT/mTOR pathway and therapeutic strategies. In: American Journal of Medical Genetics, Part C: Seminars in Medical Genetics. Wiley Online Library; 2016. p. 402–21.\nNácul AP, Lecke SB, Edelweiss MI, et al. Gene expression of leptin and long leptin receptor isoform in endometriosis: a case-control study. Obstet Gynecol Int. 2013;2013(1): 879618.\nWu MH, Chen KF, Lin SC, et al. Aberrant expression of leptin in human endometriotic stromal cells is induced by elevated levels of hypoxia inducible factor-1α. Am J Pathol. 2007;170(2):590–8.\nGonçalves HF, Zendron C, Cavalcante FS, et al. Leptin, its receptor and aromatase expression in deep infiltrating endometriosis. J Ovarian Res. 2015;8:1–7.\nAlvarez P, Bogen O, Chen X, et al. Ectopic endometrium-derived leptin produces estrogen-dependent chronic pain in a rat model of endometriosis. Neuroscience. 2014;258:111–20.\nKim TH, Yu Y, Luo L, et al. Activated AKT pathway promotes establishment of endometriosis. Endocrinology. 2014;155(5):1921–30.\nMahmoudi R, Noori Alavicheh B, Nazer Mozaffari MA, et al. Polymorphisms of leptin (-2548 G/A) and leptin receptor (Q223R) genes in iranian women with breast cancer. Int J Genom. 2015;2015(1): 132720.\nRevised American Society for Reproductive Medicine classification of endometriosis: 1996. Fertil Steril. 1997; 67(5): 817–21.\nBoumaiza I, Omezzine A, Rejeb J, et al. Relationship between leptin G2548A and leptin receptor Q223R gene polymorphisms and obesity and metabolic syndrome risk in Tunisian volunteers. Genet Test Mol Biomark. 2012;16(7):726–33.\nJahantigh D, Ghazaey Zidanloo S, Forghani F, et al. IL-27 variants might be genetic risk factors for preeclampsia: based on genetic polymorphisms, haplotypes and in silico approach. Mol Biol Rep. 2020;47(10):7929–40.\nJahantigh D, Zidanloo SG, Moossavi SZ, Forghani F. Interleukin 12B rs3212227 and rs6887695 single nucleotide polymorphisms are associated with the susceptibility to preeclampsia: genetic, haplotype and bioinformatics analysis. Cytokine. 2023;164: 156166.\nNorouzi MA, Jahantigh D, Forghani F, et al. Interleukin 17 RA and RC gene polymorphisms and increased preeclampsia risk: single and combined genetic analysis. Hum Immunol. 2025;86(2): 111250.\nBains V, Kaur H, Badaruddoza B. Association analysis of polymorphisms in LEP (rs7799039 and rs2167270) and LEPR (rs1137101) gene towards the development of type 2 diabetes in North Indian Punjabi population. Gene. 2020;754: 144846.\nMammes O, Betoulle D, Aubert R, et al. Erratum: Novel polymorphisms in the 5’ region of the LEP gene: association with leptin levels and response to low-calorie diet in human obesity (Diabetes (1998) 47 (487–489)). Diabetes. 2000;49(9):1617.\nVega MV, Obregón AM, Weisstaub G, et al. Association between feeding behavior, and genetic polymorphism of leptin and its receptor in obese chilean children. Nutr Hosp. 2015;31(3):1044–51.\nGuízar-Mendoza JM, Amador-Licona N, Flores-Martínez SE, et al. Association analysis of the Gln223Arg polymorphism in the human leptin receptor gene, and traits related to obesity in Mexican adolescents. J Hum Hypertens. 2005;19(5):341–6.\nMéar L, Herr M, Fauconnier A, et al. Polymorphisms and endometriosis: a systematic review and meta-analyses. Hum Reprod Update. 2020;26(1):73–103.\nKargasheh FB, Ansaripour S, Borumandnia N, et al. Association of leptin G2548A and leptin receptor Q223R polymorphisms and their serum levels with infertility and recurrent pregnancy loss in Iranian women with polycystic ovary syndrome. PLoS One. 2021;16(8):e0255920.\nAkman M, Karakuş N, Rüstemoğlu H. Dual effect of LEP -2548G>A (rs7799039) and LEPR Q223R (rs1137101) polymorphisms as a risk factor for azoospermic male infertility in Turkish population. Cukurova Med J. 2023;48(2):369–76.\nKhosropour S, Hamidi M, Fattahi A, et al. Leptin and leptin-receptor polymorphisms in fertile and infertile men. Syst Biol Reprod Med. 2017;63(1):7–14.\nTakemura Y, Osuga Y, Harada M, et al. Concentration of adiponectin in peritoneal fluid is decreased in women with endometriosis. Am J Reprod Immunol. 2005;54(4):217–21.\nTakemura Y, Osuga Y, Yamauchi T, et al. Expression of adiponectin receptors and its possible implication in the human endometrium. Endocrinology. 2006;147(7):3203–10.\nGonzález RR, Caballero-Campo P, Jasper M, et al. Leptin and leptin receptor are expressed in the human endometrium and endometrial leptin secretion is regulated by the human blastocyst. J Clin Endocrinol Metab. 2000;85(12):4883–8.\nKim TH, Bae N, Kim T, et al. Leptin stimulates endometriosis development in mouse models. Biomedicines. 2022;10(9):2160.\nWu MH, Chuang PC, Chen HM, et al. Increased leptin expression in endometriosis cells is associated with endometrial stromal cell proliferation and leptin gene up-regulation. Mol Hum Reprod. 2002;8(5):456–64.\nSabogal JC, Muñoz L. Leptin in obstetrics and gynecology: a review. Obstet Gynecol Surv. 2001;56(4):225–30.\nStyer AK, Sullivan BT, Puder M, et al. Ablation of leptin signaling disrupts the establishment, development, and maintenance of endometriosis-like lesions in a murine model. Endocrinology. 2008;149(2):506–14.\nAhn JH, Choi YS, Choi JH. Leptin promotes human endometriotic cell migration and invasion by up-regulating MMP-2 through the JAK2/STAT3 signaling pathway. Mol Hum Reprod. 2015;21(10):792–802.\nRathore N, Kriplani A, Yadav RK, et al. Distinct peritoneal fluid ghrelin and leptin in infertile women with endometriosis and their correlation with interleukin-6 and vascular endothelial growth factor. Gynecol Endocrinol. 2014;30(9):671–5.\nMahutte NG, Matalliotakis IM, Goumenou AG, et al. Inverse correlation between peritoneal fluid leptin concentrations and the extent of endometriosis. Hum Reprod. 2003;18(6):1205–9.\nDe Placido G, Alviggi C, Carravetta C, et al. The peritoneal fluid concentration of leptin is increased in women with peritoneal but not ovarian endometriosis. Hum Reprod. 2001;16(6):1251–4.\nAcknowledgments\nThis work is based on the Residency's Thesis Number 10338 at Zahedan University of Medical Sciences. The authors would like to thank the study participants (patients and controls) in this study.\nAuthor information\nAuthors and Affiliations\nContributions\nFF and DJ conceived and designed the experiments. DJ and MT analyzed the data. EKA and MR performed the genotyping. DJ and SGZ wrote the first draft of the manuscript. DJ performed bioinformatics analysis. FF, EKA, MR and MT contributed to the writing of the manuscript. All authors reviewed and approved the final manuscript.\nCorresponding authors\nEthics declarations\nCompeting interests\nThe authors declare that they have no competing interests.\nEthical Approval\nEthical approval was obtained from the local Ethics committee of Zahedan University of Medical Sciences (Ethical code: IR.ZAUMS.REC.1401.080), in accordance with the Declaration of Helsinki.\nAdditional information\nPublisher's Note\nSpringer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.\nRights and permissions\nSpringer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.\nAbout this article\nCite this article\nKiyani-Afzali, E., Forghani, F., Jahantigh, D. et al. Leptin G2548 A and Leptin Receptor Q223R Gene Polymorphisms are Associated with an Increased Risk of Endometriosis. Bratisl. Med. J. 126, 2024–2036 (2025). https://doi.org/10.1007/s44411-025-00186-6\nReceived:\nRevised:\nAccepted:\nPublished:\nVersion of record:\nIssue date:\nDOI: https://doi.org/10.1007/s44411-025-00186-6","source_license":"CC0","license_restricted":false}