PKOS’lu Hastalarda INSR geni Ekzom Dizileme Sonuçları

Abstract

Giriş ve Amaç: Polikistik over sendromu (PKOS), içerisinde insülin direnci, infertilite, hiperandrojenizm gibi bulgularının yer aldığı bir endokrinopatidir. Genetik alt yapısı çeşitli genlerde bulunan varyasyon ve mutasyonlarla araştırılmaya devam etmektedir. Gereç ve Yöntemler: Bu çalışmada AE-PKOS kriterlerine göre prospektif 16 PKOS hastasının klinik ve laboratuvar bulguları değerlendirip ve INSR geni ekzom dizileme yöntemi ile analiz ettik. Sonuçlar: Toplamda, 3 tanesini kendimizin isimlendirdiğimiz 4 novel varyasyonu ortaya çıkardık. Bunlar; NM_000208.4:c.974+82_974+83insT, NM_000208.4:c.974+107_974+108insC, NM_000208.4:c.653-79A>G ve rs1449625253 intronik varyasyonlarıdır. Toplamda 41 çeşit varyasyonu içeren 196 varyasyon ortaya çıkardık. INSR geninde bulduğumuz 27 bilgi verici SNP kullanılarak ilk kez bu çalışmada, yüksek linkaj eşitsizliği skoru (r2=1) gösteren 2 farklı üçerli tag SNP bloğunu saptadık. Bunlar; rs2963-rs2245649-rs2245655 ile rs6413502-rs41509747-rs73498780 bloklarıdır. Ayrıca, en sık görülen rs7508516 ve hastalarımızda da gözlemlediğimiz ve PKOS’dan sorumlu olabilecek rs2059807 gibi varyasyonların MAF değerlerini ortaya çıkardık. Tartışma: Çalışma Türkiye’de PKOS hastalarının INSR genlerinin ekzom dizileme yöntemi ile araştırıldığı ilk çalışmadır. INSR genine ait tag SNP’lerin ilk kez ortaya çıkarıldığı ve yeni aday varyasyonları bulduğumuz çalışma literatüre önemli veriler sunmaktadır.

Keywords

• PKOS
• tag SNP
• INSR geni
• linkaj eşitsizliği
• ekzom dizileme

Supporting Institution

Yozgat Bozok Üniversitesi Bilimsel Araştırma Projeleri Birimi

Project Number

6602cTF/21-469

Ethical Statement

Çalışmamız 2017-KAEK-189_2021.02.24_07 protokol numarası ile Yozgat Bozok Üniversitesi Klinik Araştırmalar Etik Kurul Onayı almıştır.

Thanks

Çalışmamız Yozgat Bozok Üniversitesi Bilimsel Araştırma Projeleri Birimi tarafından desteklenmiştir. Kurumumuza destekleri için teşekkür ederiz.

INSR gene Exome Sequencing Results in patients with PCOS

Abstract

Objective: Polycystic ovary syndrome (PCOS) is an endocrinopathy that includes findings such as insulin resistance, infertility and hyperandrogenism. Its genetic background continues to be investigated with variations and mutations in various genes. Material and Methods: In current study, we evaluated the clinical and laboratory findings of 16 prospective PCOS patients according to AE-PCOS criteria and analyzed the INSR gene by exome sequencing method. Results: We revealed 4 novel variations, 3 of which we named ourselves. These are NM_000208.4:c.974+82_974+83insT, NM_000208.4:c.974+107_974+108insC, NM_000208.4:c.653-79A>G and rs1449625253 and are all intronic variations. In this study, using the 27 informative SNPs that we found in the INSR gene, we detected two different triple tag SNP blocks showing high linkage disequilibrium score (r2 = 1) for the first time. These blocks are rs2963-rs2245649-rs2245655 and rs6413502-rs41509747-rs73498780. We also revealed the MAF values of variations such as rs7508516, which is the most common, and rs2059807, which we observed in our patients and may be responsible for PCOS. Conclusion: The study is the first study in Turkiye in which the INSR genes of PCOS patients were investigated by exome sequencing method. The study, in which the tag SNPs of the INSR gene were revealed for the first time and we found new candidate variations, provides important data to the literature.

Keywords

• PCOS
• tag SNP
• INSR gene
• linkage disequilibrium
• exome sequencing

Supporting Institution

Yozgat Bozok University Scientific Research Projects Unit.

Project Number

6602cTF/21-469

Ethical Statement

Our study has been approved by Yozgat Bozok University Clinical Research Ethics Committee Approval with protocol number 2017-KAEK-189_2021.02.24_07.

Thanks

Authors thank to supporters.

References

1. 1. Stein IF, Leventhal ML. Amenorrhea associated with bilateral polycystic ovaries. Am J Obstet Gynecol. 1935;
2. 2. Cooper HE, Spellacy WN, Prem KA, Cohen WD. Hereditary factors in the Stein-Leventhal syndrome. Am J Obstet Gynecol. 1968;
3. 3. Givens JR, Wiser WL, Coleman SA, Wilroy RS, Andersen RN, Fish SA. Familial ovarian hyperthecosis: A study of two families. Am J Obstet Gynecol. 1971;
4. 4. HAGUE WM, ADAMS J, REEDERS ST, PETO TEA, JACOBS HS. FAMILIAL POLYCYSTIC OVARIES: A GENETIC DISEASE? Clin Endocrinol (Oxf). 1988;
5. 5. Khan MJ, Ullah A, Basit S. Genetic basis of polycystic ovary syndrome (PCOS): Current perspectives. Application of Clinical Genetics. 2019.
6. 6. Ates S, Sevket O, Sudolmus S, Dane B, Ozkal F, Uysal O, et al. Different phenotypes of polycystic ovary syndrome in Turkish women: Clinical and endocrine characteristics. Gynecol Endocrinol. 2013;
7. 7. Duz S. Polycystic Ovary Syndrome [Polikistik Over Sendromu]. Med Sci | Int Med J. 2016;5(2):583.
8. 8. Gaasenbeek M, Powell BL, Sovio U, Haddad L, Gharani N, Bennett A, et al. Large-Scale Analysis of the Relationship between CYP11A Promoter Variation, Polycystic Ovarian Syndrome, and Serum Testosterone. J Clin Endocrinol Metab. 2004;

9. 9. Mifsud A. Androgen Receptor Gene CAG Trinucleotide Repeats in Anovulatory Infertility and Polycystic Ovaries. J Clin Endocrinol Metab. 2000;
10. 10. Wickham EP, Ewens KG, Legro RS, Dunaif A, Nestler JE, Strauss JF. Polymorphisms in the SHBG gene influence serum SHBG levels in women with polycystic ovary syndrome. J Clin Endocrinol Metab. 2011;
11. 11. Munir I, Yen HW, Geller DH, Torbati D, Bierden RM, Weitsman SR, et al. Insulin Augmentation of 17α-Hydroxylase Activity Is Mediated by Phosphatidyl Inositol 3-Kinase but Not Extracellular Signal-Regulated Kinase-1/ 2 in Human Ovarian Theca Cells. Endocrinology. 2004;
12. 12. Poretsky L. The Insulin-Related Ovarian Regulatory System in Health and Disease. Endocr Rev. 1999;
13. 13. Baillargeon JP, Carpentier A. Role of insulin in the hyperandrogenemia of lean women with polycystic ovary syndrome and normal insulin sensitivity. Fertil Steril. 2007;
14. 14. Kennedy GC, German MS, Rutter WJ. The minisatellite in the diabetes susceptibility locus IDDM2 regulates insulin transcription. Nat Genet. 1995;
15. 15. Paquette J, Giannoukakis N, Polychronakos C, Vafiadis P, Deal C. The INS 5’ variable number of tandem repeats is associated with IGF2 expression in humans. J Biol Chem. 1998;
16. 16. Waterworth DM, Bennett ST, Gharani N, McCarthy MI, Hague S, Batty S, et al. Linkage and association of insulin gene VNTR regulatory polymorphism with polycystic ovary syndrome. Lancet. 1997;
17. 17. Vanková M, Vrbíková J, Hill M, Cinek O, Bendlová B. Association of insulin gene VNTR polymorphism with polycystic ovary syndrome. In: Annals of the New York Academy of Sciences. 2002.
18. 18. Yun JH, Gu BH, Kang Y Bin, Choi BC, Song S, Baek KH. Association between INS-VNTR polymorphism and polycystic ovary syndrome in a Korean population. Gynecol Endocrinol. 2012;
19. 19. Conway GS, Avey C, Rumsby G. Genetics: The tyrosine kinase domain of the insulin receptor gene is normal in women with hyperinsulinaemia and polycystic ovary syndrome. Hum Reprod. 1994;
20. 20. https://gvs.gs.washington.edu/GVS150.
21. 21. https://Idlink.nih.gov/?tab=ldpop.
22. 22. Stepto NK, Cassar S, Joham AE, Hutchison SK, Harrison CL, Goldstein RF, et al. Women with polycystic ovary syndrome have intrinsic insulin resistance on euglycaemic-hyperinsulaemic clamp. Hum Reprod. 2013 Mar 1;28(3):777–84.
23. 23. Cassar S, Misso ML, Hopkins WG, Shaw CS, Teede HJ, Stepto NK. Insulin resistance in polycystic ovary syndrome: a systematic review and meta-analysis of euglycaemic–hyperinsulinaemic clamp studies. Hum Reprod. 2016 Nov;31(11):2619–31.
24. 24. Colilla S, Cox NJ, Ehrmann DA. Heritability of Insulin Secretion and Insulin Action in Women with Polycystic Ovary Syndrome and Their First Degree Relatives1. J Clin Endocrinol Metab. 2001 May 1;86(5):2027–31.
25. 25. Sir-Petermann T, Codner E, Pérez V, Echiburú B, Maliqueo M, Ladrón de Guevara A, et al. Metabolic and Reproductive Features before and during Puberty in Daughters of Women with Polycystic Ovary Syndrome. J Clin Endocrinol Metab. 2009 Jun 1;94(6):1923–30.
26. 26. Ardon O, Procter M, Tvrdik T, Longo N, Mao R. Sequencing analysis of insulin receptor defects and detection of two novel mutations in INSR gene. Mol Genet Metab reports. 2014;1:71–84.
27. 27. Dakshinamoorthy J, Jain PR, Ramamoorthy T, Ayyappan R, Balasundaram U. Association of GWAS identified INSR variants (rs2059807 & rs1799817) with polycystic ovarian syndrome in Indian women. Int J Biol Macromol. 2020 Feb;144:663–70.
28. 28. Shi Y, Zhao H, Shi Y, Cao Y, Yang D, Li Z, et al. Genome-wide association study identifies eight new risk loci for polycystic ovary syndrome. Nat Genet. 2012 Sep 12;44(9):1020–5.
29. 29. Feng C, Lv P-P, Yu T-T, Jin M, Shen J-M, Wang X, et al. The Association between Polymorphism of INSR and Polycystic Ovary Syndrome: A Meta-Analysis. Int J Mol Sci. 2015 Jan 22;16(2):2403–25.
30. 30. Bagheri M, Abdi-Rad I, Hosseini-Jazani N, Zarrin R, Nanbakhsh F, Mohammadzaie N. An Association Study between INSR/NsiI (rs2059806) and INSR/PmlI (rs1799817) SNPs in Women with Polycystic Ovary Syndrome from West Azerbaijan Province, Iran. J Reprod Infertil. 2015;16(2):109–12.
31. 31. Unsal T, Konac E, Yesilkaya E, Yilmaz A, Bideci A, Ilke Onen H, et al. Genetic polymorphisms of FSHR, CYP17, CYP1A1, CAPN10, INSR, SERPINE1 genes in adolescent girls with polycystic ovary syndrome. J Assist Reprod Genet. 2009 Apr 22;26(4):205–16.
32. 32. https://genome.ucsc.edu/cgi-bin/hgTracks?db=hg19.
33. 33. Legro RS, Arslanian SA, Ehrmann DA, Hoeger KM, Murad MH, Pasquali R, et al. Diagnosis and Treatment of Polycystic Ovary Syndrome: An Endocrine Society Clinical Practice Guideline. J Clin Endocrinol Metab. 2013 Dec;98(12):4565–92.