AMHR2 Polymorphism in Blood and Granulosa Cells in Unexplained Infertility

AMH, Infertility, Female, IVF

unexplained infertility, anti-mullerian hormone receptor2 polymorphism

There are various known parameters for assessing ovarian reserve, including ovarian volume, antral follicle count, follicle stimulating hormone (FSH) and Inhibin B at the beginning of the menstrual cycle, as well as anti-Mullerian hormone (AMH). Anti-müllerian hormone is a dimeric glycoprotein and a member of the transforming growth factor β family that plays a role in the regulation of follicular development. AMH is produced by granulosa cells of the early developing follicles in the ovary, and continues to be expressed in the growing follicles until these follicles have reached a size of 4-6 mm and a differentiation state at which AMH becomes receptive for exogenous FSH, and may be selected for dominance.It exerts its biological effects through the receptor AMHR2, which is present on granulosa and theca cells. Considering the important role of the AMH signalling pathway in regulating FSH sensitivity in the ovary and follicular recruitment and selection, it is appropriate to consider that variation in the genes encoding key proteins in the pathway may influence ovarian response. The aim of this study is to investigate the effect of the distribution of single gene polymorphisms of the AMHR2 receptor gene A-482G/rs200255 in patients with unexplained infertility and to compare this distribution with the distribution in women without infertility.

This prospective, controlled cohort study was conducted at IVF Centre of Department of Obstetrics and Gynecology, Bezmialem University Hospital between October 2022 and April 2023. The study protocol was approved by the Ethical Committee of the Medical Faculty of Bezmialem University. Written informed consent was obtained from all patients. This trial was designed and reported according to the Consolidated Standards of Reporting Trials (CONSORT) guidelines. The patients were divided into 3 groups. Group 1 (study group; unexplained infertilitiy, n=32), Group 2 ( first control group; healthy women with a history of at least one successful previous pregnancy, n=32), Group 3 ( second control group; infertile patients undergo IVF because of male or tubal factor, n=32). Inclusion criteria for the study were as follows: \<40 years of age, normal serum levels of TSH and prolactin presence of both ovaries without any morphological abnormalities, normal ovulatory cycles (25-35 days), body mass index (BMI) ≤30, no previous history of poor response and no evidence of endocrine disease, such as polycystic ovary syndrome, hipo-hyper prolactinemia, no use of hormone therapy in the 6 months preceding the recruitment and commencing IVF treatment. Patients with moderate/severe endometriosis (stage III and IV), previous ovarian surgery or underwent chemo/radiotherapy excluded from the study. At the first application, peripheral blood will be taken from the Study group, 1st Control and 2nd Control group to evaluate polymorphisms of the AMHR2-482A\>G(rs200255) with PCR analysis. During IVF treatment at the day of ovum pick up; follicular fluid will be collected to isolate granulosa cell for polymorphisms of the AMHR2-482A\>G(rs200255) with PCR analysis. The total number of embryos, the number of embryos transferred, the number of frozen and viable embryos will be recorded. After the embryo transfer is performed in the study group and the 2nd control group, the pregnancy results will be recorded by checking the bhcg in the blood on the 10-12th day of the transfer.

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