Analysis of the pathogenic CYP21A2 gene variants in patients with clinical, biochemical and combined manifestations of hyperandrogenism

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Abstract

BACKGROUND: The association of heterozygous carriage of pathogenic variants in the CYP21A2 gene with various manifestations of hyperandrogenism remains poorly understood to date.

AIM: The aim of this study was to analyze the relationship between the carriage of pathogenic variants in the CYP21A2 gene in women and various manifestations of hyperandrogenism.

MATERIALS AND METHODS: Clinical description, hormonal testing and molecular genetic analysis of the CYP21A2 gene were performed in 97 women with clinical, biochemical and combined manifestations of hyperandrogenism and in 46 people in the control group. The mean age of the patients was 27.3 ± 0.6 years. Levels of 17-hydroxyprogesterone, dehydroepiandrosterone sulfate and androstenedione were measured in the blood serum of the study participants. To identify pathogenic variants in the CYP21A2 gene, we used next generation sequencing, restriction fragment length polymorphism analysis, real-time polymerase chain reaction, and multiplex ligation-dependent probe amplification analysis. A statistical analysis of the frequency of pathogenic CYP21A2 gene variants in the study groups and hormone levels in different study subgroups was carried out.

RESULTS: In patients with hirsutism, acne, menstrual irregularity, miscarriage and infertility, pathogenic variants in the CYP21A2 gene were identified in 31% (30/97) of cases in the heterozygous state and in 6% (5/97) of cases in the homozygous state. The frequency of these variants (in the heterozygous state only) was significantly higher — 6.5% (3/46) of cases, when compared to the control group (p < 0.0001). The identified pathogenic variants included both single nucleotide substitutions such as P31L (n = 1), I2splice (n = 1), V282L (n = 15), I173N (n = 3), Q319X (n = 8), R357W (n = 1), P454S (n = 1), and P483S (n = 1) and deletions of various lengths (n = 10). We found no significant difference in the levels of dehydroepiandrosterone sulfate and androstenedione between heterozygous carriers and the control group and between heterozygous carriers and patients with the wild type CYP21A2 gene (p > 0.05), while 17-hydroxyprogesterone level in the group of patients with heterozygous changes was higher, when compared to the control group (p < 0.001).

CONCLUSIONS: Heterozygous carriage of pathogenic variants in the CYP21A2 gene is associated with manifestations of hyperandrogenism. However, further study of the mechanisms underlying this association is required.

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About the authors

Natalia S. Osinovskaya

The Research Institute of Obstetrics, Gynecology and Reproductology named after D.O. Ott; North-Western State Medical University named after I.I. Mechnikov

Author for correspondence.
Email: natosinovskaya@mail.ru
ORCID iD: 0000-0001-7831-9327
SPIN-code: 3190-2307
ResearcherId: K-1168-2018

Cand. Sci. (Biol.)

Russian Federation, 3 Mendeleevskaya Line, Saint Petersburg, 199034; Saint Petersburg

Olga B. Glavnova

The Research Institute of Obstetrics, Gynecology and Reproductology named after D.O. Ott

Email: o.glavnova@mail.ru
SPIN-code: 8040-5425

Doctor endocrinologist, Head of the department of Endocrinology

Russian Federation, 3 Mendeleevskaya Line, Saint Petersburg, 199034

Maria I. Yarmolinskaya

The Research Institute of Obstetrics, Gynecology and Reproductology named after D.O. Ott; North-Western State Medical University named after I.I. Mechnikov

Email: m.yarmolinskaya@gmail.com
ORCID iD: 0000-0002-6551-4147
SPIN-code: 3686-3605
ResearcherId: P-2183-2014

MD, Dr. Sci. (Med.), Professor, Professor of the Russian Academy of Sciences

Russian Federation, 3 Mendeleevskaya Line, Saint Petersburg, 199034; Saint Petersburg

Iskander Yu. Sultanov

The Research Institute of Obstetrics, Gynecology and Reproductology named after D.O. Ott

Email: timbuctu@mail.ru

biologist of the Laboratory of Genomics with the Bioresource Collection Group of the Department of Genomic Medicine

Russian Federation, 3 Mendeleevskaya Line, Saint Petersburg, 199034

Dmitry Yu. Klyuchnikov

Dynasty, Samara Regional Medical Center

Email: dmklyu@gmail.com
ORCID iD: 0000-0003-4934-5619
SPIN-code: 4490-5989

Biologist in the Tissue Typing Laboratory

Russian Federation, Samara

Natalia N. Tkachenko

The Research Institute of Obstetrics, Gynecology and Reproductology named after D.O. Ott

Email: liberin@mail.ru
ORCID iD: 0000-0001-6189-3488
SPIN-code: 9633-6701
ResearcherId: K-1734-2018

Cand. Sci. (Biol.)

Russian Federation, 3 Mendeleevskaya Line, Saint Petersburg, 199034

Yulia A. Nasykhova

The Research Institute of Obstetrics, Gynecology and Reproductology named after D.O. Ott

Email: yulnasa@gmail.com
ORCID iD: 0000-0002-3543-4963
SPIN-code: 9661-9416

Cand. Sci. (Biol.)

Russian Federation, 3 Mendeleevskaya Line, Saint Petersburg, 199034

Andrey S. Glotov

The Research Institute of Obstetrics, Gynecology and Reproductology named after D.O. Ott

Email: anglotov@mail.ru
ORCID iD: 0000-0002-7465-4504
SPIN-code: 1406-0090
ResearcherId: E-8525-2015

Dr. Sci. (Biol.)

Russian Federation, 3 Mendeleevskaya Line, Saint Petersburg, 199034

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Supplementary files

Supplementary Files
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1. Fig. 1. An example of a detected complete deletion of the CYP21A2 gene in the heterozygous state (the arrows show the beginning and end of the deleted sequence). The areas of gene analysis: CYP21A1P, CYP21A2, TNXB, reference gene is marked on the x-axis

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2. Fig. 2. The frequency of detection of menstrual irregularity, acne, miscarriage, primary infertility and hirsutism in heterozygous carriers of the pathogenic CYP21A2 gene variants and in patients with the wild type, as well as in separate subgroups of patients — with and without the V282L variant

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3. Fig. 3. The frequency of heterozygous carriers of the pathogenic CYP21A2 gene variants in subgroups of patients with menstrual irregularity, acne, miscarriage, primary infertility and hirsutism

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