Genes & Genomics

, Volume 41, Issue 1, pp 61–70 | Cite as

Association study of the three functional polymorphisms (TAS2R46G>A, OR4C16G>A, and OR4X1A>T) with recurrent pregnancy loss

  • Chang Soo Ryu
  • Jung Hyun Sakong
  • Eun Hee Ahn
  • Jung Oh Kim
  • Daeun Ko
  • Ji Hyang Kim
  • Woo Sik Lee
  • Nam Keun KimEmail author
Research Article


This study was purposed to investigate whether genetic polymorphisms in the function of stop-gain are associated with a fetal or placental development play roles and a development of idiopathic recurrent pregnancy loss (RPL) in Korean females. Three stop-gain polymorphisms were selected using next-generation sequencing screening, which allows for the rigorous examination and discovery of previously uncharacterized stop-gain genes and stop-gain expression profiles. Accordingly, we investigated the association of stop-gain polymorphisms in Korean women with RPL. Three functional polymorphisms in the TAS2R46G>A (rs2708381), OR4C16G>A (rs1459101), and OR4X1A>T (rs10838851) genes were genotyped using polymerase chain reaction (PCR)—restriction fragment length polymorphism assays and real-time PCR analysis. We determined that the OR4C16G>A polymorphism was associated with idiopathic RPL in Korean women (Adjusted odds ratio [AOR] 1.782; 95% confidence interval [CI] 1.004–3.163; P = 0.048, and AOR 1.766; 95% CI 1.020–3.059; P = 0.042). In addition, the prevalence of RPL was increased in women with the OR4C16GA + AA genotype and blood coagulation measures of prothrombin time (PT) > 10.4 s (AOR 8.292; 95% CI 2.744–25.054). We suggest that the OR4C16G>A polymorphism might serve as a clinically useful biomarker for the development, prevention, and prognosis of RPL.


Next-generation sequencing (NGS) Stop gain mutation Nonsense mutation Polymorphism Recurrent pregnancy loss (RPL) 



This study was partly supported by a grant of the Korea Healthcare Technology R&D Project (HI15C1972010015), Ministry for Health, Welfare & Family Affairs, Republic of Korea.

Author contributions

CSR and NKK designed and directed the whole project. EHA, JHK, and WSL collected the blood samples from recurrent pregnancy loss patients and control subjects. CSR and JHS performed the experiments, collected the results, and analyzed the data. NKK and EHA discussed and interpreted the data and results. CSR and JHS wrote the first draft of the manuscript. NKK, JOK, DK and CSR revised the paper. All authors contributed to and approved the final manuscript.

Compliance with ethical standards

Conflict of interest

Chang Soo Ryu, Jung Hyun Sakong, Eun Hee Ahn, Jung Oh Kim, Daeun Ko, Ji Hyang Kim, Woo Sik Lee and Nam Keun Kim declare that they have no conflict of interest.

Ethical approval

This study had been approved by the Institutional Review Board of CHA Bundang Medical Center (IRB number: BD2010‑123D) and written informed consent was provided by all patients.

Supplementary material

13258_2018_738_MOESM1_ESM.docx (324 kb)
Supplementary material 1 (DOCX 324 KB)


  1. Abel BS, Shaw ND, Brown JM, Adams JM, Alati T, Martin KA, Pitteloud N, Seminara SB, Plummer L, Pignatelli D et al (2013) Responsiveness to a physiological regimen of GnRH therapy and relation to genotype in women with isolated hypogonadotropic hypogonadism. J Clin Endocrinol Metab 98:E206–E216CrossRefGoogle Scholar
  2. Ahn YJ, Markkandan K, Baek IP, Mun S, Lee W, Kim HS, Han K (2018) An efficient and tunable parameter to improve variant calling for whole genome and exome sequencing data. Genes Genom 40:39–47CrossRefGoogle Scholar
  3. Benjamini Y, Drai D, Elmer G, Kafkafi N, Golani I (2001) Controlling the false discovery rate in behavior genetics research. Behav Brain Res 125:279–284CrossRefGoogle Scholar
  4. Bhuiyan ZA, Momenah TS, Gong Q, Amin AS, Ghamdi SA, Carvalho JS, Homfray T, Mannens MM, Zhou Z, Wilde AA et al (2008) Recurrent intrauterine fetal loss due to near absence of HERG: clinical and functional characterization of a homozygous nonsense HERG Q1070X mutation. Heart Rhythm 5:553–561CrossRefGoogle Scholar
  5. Bondeson ML, Ericson K, Gudmundsson S, Ameur A, Ponten F, Wesstrom J, Frykholm C, Wilbe M (2017) A nonsense mutation in CEP55 defines a new locus for a Meckel-like syndrome, an autosomal recessive lethal fetal ciliopathy. Clin Genet 92:510–516CrossRefGoogle Scholar
  6. Buchholz T, Lohse P, Rogenhofer N, Kosian E, Pihusch R, Thaler CJ (2003) Polymorphisms in the ACE and PAI-1 genes are associated with recurrent spontaneous miscarriages. Hum Reprod 18:2473–2477CrossRefGoogle Scholar
  7. Cameron EL (2014) Pregnancy and olfaction: a review. Front Psychol 5:67Google Scholar
  8. Carbone T, Nasorri F, Pennino D, Eyerich K, Foerster S, Cifaldi L, Traidl-Hoffman C, Behrendt H, Cavani A (2010) CD56highCD16-CD62L- NK cells accumulate in allergic contact dermatitis and contribute to the expression of allergic responses. J Immunol 184:1102–1110CrossRefGoogle Scholar
  9. Chandrashekar J, Mueller KL, Hoon MA, Adler E, Feng L, Guo W, Zuker CS, Ryba NJ (2000) T2Rs function as bitter taste receptors. Cell 100:703–711CrossRefGoogle Scholar
  10. Chihara M, Yoshihara K, Ishiguro T, Yokota Y, Adachi S, Okada H, Kashima K, Sato T, Tanaka A, Tanaka K et al (2015) Susceptibility to male infertility: replication study in Japanese men looking for an association with four GWAS-derived loci identified in European men. J Assist Reprod Genet 32:903–908CrossRefGoogle Scholar
  11. Choi YS, Kwon H, Kim JH, Shin JE, Choi Y, Yoon TK, Choi DH, Kim NK (2011) Haplotype-based association of ACE I/D, AT1R 1166A>C, and AGT M235T polymorphisms in renin-angiotensin-aldosterone system genes in Korean women with idiopathic recurrent spontaneous abortions. Eur J Obstet Gynecol Reprod Biol 158:225–228CrossRefGoogle Scholar
  12. Fauchereau F, Shalev S, Chervinsky E, Beck-Fruchter R, Legois B, Fellous M, Caburet S, Veitia RA (2016) A non-sense MCM9 mutation in a familial case of primary ovarian insufficiency. Clin Genet 89:603–607CrossRefGoogle Scholar
  13. Fu W, O’Connor TD, Jun G, Kang HM, Abecasis G, Leal SM, Gabriel S, Rieder MJ, Altshuler D, Shendure J et al (2013) Analysis of 6515 exomes reveals the recent origin of most human protein-coding variants. Nature 493:216–220CrossRefGoogle Scholar
  14. Gaillard I, Rouquier S, Giorgi D (2004) Olfactory receptors. Cell Mol Life Sci 61:456–469CrossRefGoogle Scholar
  15. Hahn LW, Ritchie MD, Moore JH (2003) Multifactor dimensionality reduction software for detecting gene-gene and gene-environment interactions. Bioinformatics 19:376–382CrossRefGoogle Scholar
  16. Hummel T, Landis BN, Huttenbrink KB (2011) Smell and taste disorders. GMS Curr Top Otorhinolaryngol Head Neck Surg 10:Doc04Google Scholar
  17. Jung YW, Jeon YJ, Rah H, Kim JH, Shin JE, Choi DH, Cha SH, Kim NK (2014) Genetic variants in microRNA machinery genes are associated [corrected] with idiopathic recurrent pregnancy loss risk. PLoS ONE 9:e95803CrossRefGoogle Scholar
  18. Kim JH, Jeon YJ, Lee BE, Kang H, Shin JE, Choi DH, Lee WS, Kim NK (2013) Association of methionine synthase and thymidylate synthase genetic polymorphisms with idiopathic recurrent pregnancy loss. Fertil Steril 99:1674–1680CrossRefGoogle Scholar
  19. Kim KS, Egan JM, Jang HJ (2014) Denatonium induces secretion of glucagon-like peptide-1 through activation of bitter taste receptor pathways. Diabetologia 57:2117–2125CrossRefGoogle Scholar
  20. Kim KS, Lee IS, Kim KH, Park J, Kim Y, Choi JH, Choi JS, Jang HJ (2017) Activation of intestinal olfactory receptor stimulates glucagon-like peptide-1 secretion in enteroendocrine cells and attenuates hyperglycemia in type 2 diabetic mice. Sci Rep 7:13978CrossRefGoogle Scholar
  21. Macarthur DG, Tyler-Smith C (2010) Loss-of-function variants in the genomes of healthy humans. Hum Mol Genet 19:R125–R130CrossRefGoogle Scholar
  22. Malnic B, Godfrey PA, Buck LB (2004) The human olfactory receptor gene family. Proc Natl Acad Sci USA 101:2584–2589CrossRefGoogle Scholar
  23. Meczekalski B, Podfigurna-Stopa A, Smolarczyk R, Katulski K, Genazzani AR (2013) Kallmann syndrome in women: from genes to diagnosis and treatment. Gynecol Endocrinol 29:296–300CrossRefGoogle Scholar
  24. Meyerhof W, Batram C, Kuhn C, Brockhoff A, Chudoba E, Bufe B, Appendino G, Behrens M (2010) The molecular receptive ranges of human TAS2R bitter taste receptors. Chem Senses 35:157–170CrossRefGoogle Scholar
  25. Moore JH, Williams SM (2002) New strategies for identifying gene-gene interactions in hypertension. Ann Med 34:88–95CrossRefGoogle Scholar
  26. Preston FE, Rosendaal FR, Walker ID, Briet E, Berntorp E, Conard J, Fontcuberta J, Makris M, Mariani G, Noteboom W et al (1996) Increased fetal loss in women with heritable thrombophilia. Lancet 348:913–916CrossRefGoogle Scholar
  27. Pronin AN, Xu H, Tang H, Zhang L, Li Q, Li X (2007) Specific alleles of bitter receptor genes influence human sensitivity to the bitterness of aloin and saccharin. Curr Biol 17:1403–1408CrossRefGoogle Scholar
  28. Qiao Y, Wen J, Tang F, Martell S, Shomer N, Leung PC, Stephenson MD, Rajcan-Separovic E (2016) Whole exome sequencing in recurrent early pregnancy loss. Mol Hum Reprod 22:364–372CrossRefGoogle Scholar
  29. Rausell A, Mohammadi P, McLaren PJ, Bartha I, Xenarios I, Fellay J, Telenti A (2014) Analysis of stop-gain and frameshift variants in human innate immunity genes. PLoS Comput Biol 10:e1003757CrossRefGoogle Scholar
  30. Ritchie MD, Hahn LW, Roodi N, Bailey LR, Dupont WD, Parl FF, Moore JH (2001) Multifactor-dimensionality reduction reveals high-order interactions among estrogen-metabolism genes in sporadic breast cancer. Am J Hum Genet 69:138–147CrossRefGoogle Scholar
  31. Rosen HR, Doherty DG, Madrigal-Estebas L, O’Farrelly C, Golden-Mason L (2008) Pretransplantation CD56(+) innate lymphocyte populations associated with severity of hepatitis C virus recurrence. Liver Transpl 14:31–40CrossRefGoogle Scholar
  32. Saxena P, Misro MM, Roy S, Chopra K, Sinha D, Nandan D, Trivedi SS (2008) Possible role of male factors in recurrent pregnancy loss. Indian J Physiol Pharmacol 52:274–282Google Scholar
  33. Siasi E, Aleyasin A (2016) Four single nucleotide polymorphisms in INSR, SLC6A14, TAS2R38, and OR2W3 genes in Association with Idiopathic Infertility in Persian Men. J Reprod Med 61:145–152Google Scholar
  34. Upadhyaya JD, Chakraborty R, Shaik FA, Jaggupilli A, Bhullar RP, Chelikani P (2016) The pharmacochaperone activity of quinine on bitter taste receptors. PLoS ONE 11:e0156347CrossRefGoogle Scholar
  35. Wang H, Wu S, Wu J, Sun S, Wu S, Bao W (2018) Association analysis of the SNP (rs345476947) in the FUT2 gene with the production and reproductive traits in pigs. Genes Genom 40:199–206CrossRefGoogle Scholar
  36. Wilcox AJ, Weinberg CR, O’Connor JF, Baird DD, Schlatterer JP, Canfield RE, Armstrong EG, Nisula BC (1988) Incidence of early loss of pregnancy. N Engl J Med 319:189–194CrossRefGoogle Scholar

Copyright information

© The Genetics Society of Korea and Springer Nature B.V. 2018

Authors and Affiliations

  • Chang Soo Ryu
    • 1
  • Jung Hyun Sakong
    • 1
  • Eun Hee Ahn
    • 2
  • Jung Oh Kim
    • 1
  • Daeun Ko
    • 4
  • Ji Hyang Kim
    • 2
  • Woo Sik Lee
    • 3
  • Nam Keun Kim
    • 1
    Email author
  1. 1.Department of Biomedical Science, College of Life ScienceCHA UniversitySeongnamSouth Korea
  2. 2.Department of Obstetrics and Gynecology, CHA Bundang Medical CenterCHA UniversitySeongnamSouth Korea
  3. 3.Fertility Center of CHA Gangnam Medical CenterCHA UniversityGangnamSouth Korea
  4. 4.Department of Anesthesiology and Pain Medicine, CHA Bundang Medical CenterCHA UniversitySeongnamSouth Korea

Personalised recommendations