Embryo implantation is a complex process that requires sequential steps at the interface of embryo interaction with decidual endometrium. Many women after experiencing multiple attempts of assisted reproductive techniques fail to get implantation because of instability of leukaemia inhibitory factor and leukaemia inhibitory factor receptor-signal transducer and activator of transcription factor 3 (LIF-LIFR STAT3) signalling cascade. Therefore, this study explores the association of ovarian steroids, LIF and LIFR stop-gained variant using the tetra primer amplification refractory mutation system-polymerase chain reaction (TARMS-PCR) with unexplained infertility (UEX-IF) among Pakistani women.

Materials and Methods

This is a case-control study, a total of 81 unexplained infertile women and 162 fertile controls (with age and BMI matched) were inducted. Serum estradiol, progesterone and LIF were determined using enzyme-linked immunosorbent assay (ELISA). T-ARMS-PCR was designed using Primer 1 software. Genomic DNA was extracted from peripheral blood and amplified using T-ARMS-PCR followed by sequencing for validation and comprehensive concordance.

Results

This study established differences in LIF levels (χ² = 9.857, P < .05) between patients and controls as well as explored the decreased LIF significantly raised the risk of UEX-IF (OR = 2.316; 95% CI = 1.214, 4.416). Progesterone (P) was significantly associated with UEX-IF between fertile and infertile counterparts (χ² = 20.347, P < .05). It was also observed that increased Progesterone reduced the risk of UEX-IF (OR = 0.306; 95% CI = 0.166, 0.567). A rapid and inexpensive method for genotyping novel LIFR gene polymorphism through T- ARMS-PCR was successfully developed. LIFR gene SNP (rs121912501) had significant association (χ² = 200.681, P < .05) with UEX-IF. LIFR rs121912501 “TT” genotype (OR = 5.417; 95% CI = 1.868, 15.709) and “CT” genotype (OR = 3.104, 95% CI = 1.586,6.076) were at increased risk of infertility.

Conclusion

UEX-IF can be caused by LIFR gene variation irrespective of increased P. It may open the doors for the discovery of new management plans for infertile women.

DISCLOSURES

The authors have declared no conflict of interest.

Open Research

DATA AVAILABILITY STATEMENT

All relevant data are available in this manuscript.

REFERENCES

1Aghajanova L, Altmäe S, Bjuresten K, et al. Disturbances in the LIF pathway in the endometrium among women with unexplained infertility. Fertil Steril. 2009; 91: 2602-2610.
10.1016/j.fertnstert.2008.04.010
CAS PubMed Web of Science® Google Scholar
2Mojarrad M, Hassanzadeh-Nazarabadi M, Tafazoli N. Polymorphism of genes and implantation failure. Int J Mol Cell Med. 2013; 2: 1.
CAS PubMed Google Scholar
3Qadi AA, Taberlay PC, Phillips JL, et al. The leukemia inhibitory factor receptor gene is a direct target of RUNX1. J Cell Biochem. 2016; 117: 49-58.
10.1002/jcb.25246
CAS PubMed Web of Science® Google Scholar
4Tapia-Pizarro A, Figueroa P, Brito J, et al. Endometrial gene expression reveals compromised progesterone signaling in women refractory to embryo implantation. Reprod Biol Endocrinol. 2014; 12: 92.
10.1186/1477-7827-12-92
CAS PubMed Web of Science® Google Scholar
5Saidah TK, Amaral WNaves, Evangelista PG, et al. Pre-implantation genetic study and its importance for fertilization in the prevention of genetic disorders. Human Reprod Archives. 2018; 33(1): 17.
10.4322/hra.001017
Google Scholar
6Yue X, Wu L, Hu W. The regulation of leukemia inhibitory factor. Cancer Cell Microenviron. 2015; 2(3): 1-5.
Google Scholar
7Dimitriadis E, Nie G, Hannan NJ, et al. Local regulation of implantation at the human fetal-maternal interface. Int J Develop Biol. 2010; 54: 313-322.
10.1387/ijdb.082772ed
CAS PubMed Web of Science® Google Scholar
8Singh M, Chaudhry P, Asselin E. Bridging endometrial receptivity and implantation: network of hormones, cytokines, and growth factors. J Endocrinol. 2011; 210: 5-14.
10.1530/JOE-10-0461
CAS PubMed Web of Science® Google Scholar
9Salleh N, Giribabu N. Leukemia inhibitory factor: roles in embryo implantation and in nonhormonal contraception. Sci World J. 2014; 1-10.
Google Scholar
10Chu B, Zhong L, Dou S, et al. miRNA-181 regulates embryo implantation in mice through targeting leukemia inhibitory factor. J Mol Cell Biol. 2015; 7: 12-22.
10.1093/jmcb/mjv006
CAS PubMed Web of Science® Google Scholar
11Tu Z, Ran H, Zhang S, et al. Molecular determinants of uterine receptivity. Int J Dev Biol. 2014; 58: 147-154.
10.1387/ijdb.130345wh
CAS PubMed Web of Science® Google Scholar
12Terakawa J, Wakitani S, Sugiyama M, et al. Embryo implantation is blocked by intraperitoneal injection with anti-LIF antibody in mice. J Reprod Develop. 2011; 57: 700-707.
10.1262/jrd.11-048H
CAS PubMed Web of Science® Google Scholar
13Altmäe S, Martínez-Conejero JA, Salumets A, et al. Endometrial gene expression analysis at the time of embryo implantation in women with unexplained infertility. Mol Hum Reprod. 2010; 16(3): 178-187.
10.1093/molehr/gap102
CAS PubMed Web of Science® Google Scholar
14Medrano RFV, de Oliveira CA. Guidelines for the tetra-primer ARMS–PCR technique development. Mol Biotechnol. 2014; 56: 599-608.
CAS PubMed Web of Science® Google Scholar
15Ahlawat S, Sharma R, Maitra A, et al. Designing, optimization and validation of tetra-primer ARMS PCR protocol for genotyping mutations in caprine Fec genes. Meta gene. 2014; 2: 439-449.
10.1016/j.mgene.2014.05.004
PubMed Google Scholar
16Baird DT, Fraser IS. Blood production and ovarian secretion rates of estradiol-17β and estrone in women throughout the menstrual cycle. J Clin Endocrinol Metab. 1974; 38: 1009-1017.
10.1210/jcem-38-6-1009
CAS PubMed Web of Science® Google Scholar
17Ibraheam IA, Ahmad MBS. Low cost non-hazardous modified extraction method of DNA from human blood. Med J Babylon. 2008; 5: 152-155.
Google Scholar
18Hambartsoumian E. Endometrial leukemia inhibitory factor (LIF) as a possible cause of unexplained infertility and multiple failures of implantation. Am J Reprod Immunol. 1998; 39: 137-143.
10.1111/j.1600-0897.1998.tb00345.x
CAS PubMed Web of Science® Google Scholar
19Laird SM, Tuckerman EM, Dalton CF, et al. The production of leukaemia inhibitory factor by human endometrium: presence in uterine flushings and production by cells in culture. Hum Reprod. 1997; 12: 569-574.
10.1093/humrep/12.3.569
CAS PubMed Web of Science® Google Scholar
20Mikolajczyk M, Wirstlein P, Skrzypczak J. Leukaemia inhibitory factor and interleukin 11 levels in uterine flushings of infertile patients with endometriosis. Hum Reprod. 2006; 21: 3054-3058.
10.1093/humrep/del225
CAS PubMed Web of Science® Google Scholar
21Margioula-Siarkou C, Prapas Y, Petousis S, et al. LIF endometrial expression is impaired in women with unexplained infertility while LIF-R expression in all infertility sub-groups. Cytokine. 2017; 96: 166-172.
10.1016/j.cyto.2017.04.009
CAS PubMed Web of Science® Google Scholar
22Fox C, Morin S, Jeong J-W, et al. Local and systemic factors and implantation: what is the evidence? Fertil Steril. 2016; 105: 873-884.
10.1016/j.fertnstert.2016.02.018
PubMed Web of Science® Google Scholar
23Pawar S, Hantak AM, Bagchi IC, et al. Minireview: Steroid-regulated paracrine mechanisms controlling implantation. Mol Endocrinol. 2014; 28: 1408-1422.
10.1210/me.2014-1074
CAS PubMed Web of Science® Google Scholar
24Moberg C, Bourlev V, Ilyasova N, et al. Endometrial expression of LIF and its receptor and peritoneal fluid levels of IL-1α and IL-6 in women with endometriosis are associated with the probability of pregnancy. Arch Gynecol Obstet. 2015; 292: 429-437.
10.1007/s00404-015-3626-0
CAS PubMed Web of Science® Google Scholar
25Wu M, Yin Y, Zhao M, et al. The low expression of leukemia inhibitory factor in endometrium: possible relevant to unexplained infertility with multiple implantation failures. Cytokine. 2013; 62: 334-339.
10.1016/j.cyto.2013.03.002
CAS PubMed Web of Science® Google Scholar
26Salamonsen LA, Menkhorst E, Dimitriadis E. Leukemia inhibitory factor and human endometrial receptivity. Indian J Physiol Pharmacol. 2010; 54: 17-26.
CAS Google Scholar
27Bhurke AS, Bagchi IC, Bagchi MK. Progesterone -regulated endometrial factors controlling implantation. Am J Reprod Immunol. 2016; 75: 237-245.
10.1111/aji.12473
CAS PubMed Web of Science® Google Scholar
28Salamonsen LA, Evans J, Nguyen H, et al. The microenvironment of human implantation: determinant of reproductive success. Am J Reprod Immunol. 2016; 75: 218-225.
10.1111/aji.12450
PubMed Web of Science® Google Scholar
29Okayama N, Fujimura K, Nakamura J, et al. Evaluation of a new efficient procedure for single nucleotide polymorphism genotyping: tetra-primer amplification refractory mutation system-polymerase chain reaction. Clin Chem Lab Med. 2004; 42: 13-16.
10.1515/CCLM.2004.004
CAS PubMed Web of Science® Google Scholar
30Wånggren K, Lalitkumar P, Hambiliki F, et al. Leukaemia inhibitory factor receptor and gp130 in the human Fallopian tube and endometrium before and after mifepristone treatment and in the human preimplantation embryo. Mol Hum Reprod. 2007; 13: 391-397.
10.1093/molehr/gam013
CAS PubMed Web of Science® Google Scholar
31Tawfeek MA, Eid MA, Hasan AM, et al. Assessment of leukemia inhibitory factor and glycoprotein 130 expression in endometrium and uterine flushing: a possible diagnostic tool for impaired fertility. BMC Womens Health. 2012; 12: 10.
10.1186/1472-6874-12-10
CAS PubMed Web of Science® Google Scholar

Citing Literature

All articles

Association of novel stop-gained leukaemia inhibitory factor receptor gene (rs121912501) variant, leukaemia inhibitory factor and ovarian steroids with unexplained infertility among Pakistani women

Abstract

Aims and Objectives

Materials and Methods

Results

Conclusion

DISCLOSURES

Open Research

DATA AVAILABILITY STATEMENT

REFERENCES

Citing Literature

References

Information

About Wiley Online Library

Help & Support

Opportunities

Connect with Wiley

Association of novel stop-gained leukaemia inhibitory factor receptor gene (rs121912501) variant, leukaemia inhibitory factor and ovarian steroids with unexplained infertility among Pakistani women

Abstract

Aims and Objectives

Materials and Methods

Results

Conclusion

DISCLOSURES

Open Research

DATA AVAILABILITY STATEMENT

REFERENCES

Citing Literature

References

Related

Information