RU2816705C1 - Panel for determining predisposition to developing disorders of steroid hormone metabolism and its use - Google Patents

Abstract

FIELD: biotechnology; medicine.

SUBSTANCE: group of inventions relates to biotechnology and medicine, namely to endocrinology and genetics. Described is a panel for determining a predisposition to developing disorders of steroid hormone metabolism based on the use of a unique combination of genotypes of polymorphisms in genes of estradiol receptors (ESR1,2), progesterone (PGR), androgen receptor (AR), in genes of steroid hormone carrier proteins, as well as in genes of enzymes catechol-O-methyltransferase (COMT), sulphotransferase 2A1, and cytochrome CYP2C9 genes. Described is a method for determining predisposition to developing disorders of steroid hormone metabolism, involving the use of a panel. Combination of polymorphisms enables simultaneous detection of the most common polymorphisms, provides a more complete diagnostic coverage and does not miss possible changes in the genotype.

EFFECT: invention can be used to determine the probability of developing functional disorders of steroid hormone metabolism, aetiologically caused by various polymorphic variants in genes responsible for enzyme activity, which are involved in biosynthesis, intracellular action and utilization of steroid hormones, as well as in steroid hormone receptor genes and proteins providing steroid hormone transport.

2 cl, 18 tbl, 8 ex

Description

Field of technology to which the invention relates

The group of inventions relates to the field of medicine, namely to endocrinology and genetics, and can be used to determine the likelihood of developing functional disorders of the metabolism of steroid hormones, etiologically caused by various polymorphic variants in the genes responsible for the activity of enzymes that are involved in biosynthesis, intracellular action and utilization steroid hormones, as well as in the genes of steroid hormone receptors and proteins that ensure the transport of steroid hormones.

State of the art

The role of steroid hormones in the body is extremely diverse - these hormones perform a total of more than 200 functions. Therefore, both the overproduction of these steroids and the lack of steroids will have a negative impact on the body, and the correct prescription of steroid hormone replacement therapy must be carried out in conjunction with the analysis of polymorphic variants in the genes for steroid hormone metabolism.

Steroid hormones are signaling molecules, part of the mechanism of humoral regulation of various processes in a multicellular organism, which is not able to exist without intercellular communication. The process of steroidogenesis is completely dependent on the work of several groups of enzymes localized in mitochondria and the smooth endoplasmic reticulum, i.e. It is possible to influence steroidogenesis by influencing these enzymes, and therefore, in which polymorphic variant the gene for the enzyme involved in steroidogenesis is located, a certain therapeutic effect can be expected.

The leading role in the formation of complexes with some steroid hormones belongs to sex steroid-binding globulin (SHBG) - it binds and, therefore, inactivates approximately 78–80% of these hormones. Information about polymorphic variants of the SHBG gene can provide prognostic information about changes in the level of this transport protein in the blood.

Steroid hormones exert their effects primarily through the classical nuclear receptor, which changes gene transcription. In addition, steroid hormones have rapid non-genomic effects as a result of interaction with surface receptors localized in the plasma membrane. Thus, the assessment of polymorphic variants of genes in steroid hormone receptors provides prognostic information about the sensitivity of steroid receptors.

As our understanding of the metabolism of steroidogenesis increases, the possibilities for applying this knowledge to clinical practice are expanding. This will make it possible to select therapy for treatment of disorders of steroidogenesis based on genetic data, according to all the principles of preventive and predictive medicine.

Currently, steroidogenesis is assessed based on the results of laboratory determination of the level of steroid hormones and SHBG, which, in combination with other clinical and laboratory indicators, determine further tactics for patient management. Based on the results of a hormonal examination, a conclusion is made about disturbances in steroidogenesis and certain hormonal therapy is prescribed. In some cases, disturbances in the metabolism of steroid hormones in the body are caused by genetic disorders in the genes of estradiol receptors (ESR1,2), progesterone (PGR), androgen receptor (AR), in the genes of steroid hormone transport proteins, as well as in the genes of catechol-O-methyltransferase enzymes (COMT), sulfotransferase 2A1, and cytochrome CYP2C9 genes.

Genetic variants in sex hormone binding globulin are known to affect blood testosterone levels. In [Ohlsson C, Wallaschofski H, Lunetta KL, Stolk L, Perry JRB, et al. (2011)] studied 14,429 men and showed that genetic variants at the sex hormone binding globulin (SHBG) locus are associated with significant variations in serum testosterone concentrations and an increased risk of low testosterone levels. The associations found can be used to better understand the pathogenesis of diseases associated with low testosterone levels.

In the article [SNPs Associated With Testosterone Levels Influence Human Facial Morphology. Front. Genet. 9:497. doi: 10.3389/fgene.2018.00497] also wrote about three SNPs associated with testosterone levels and influence on facial shape and size (rs12150660, rs1799941, rs8023580), and indicated that testosterone-related genetic variants determine normal facial morphology.

An analogue of the stated solution is the panel “Analysis of polymorphisms in genes encoding enzymes of sex hormone metabolism - optimal”: AR, SHBG, INS, COMP, PGR, RRARG, SRD5A2, CYP17A1, CYP19A1, ESR1, ESR2 found on the Internet 01/25/2024 https:/ /dnkom.ru/upload/iblock/bfb/kkwp52gpb0p51twhhda2d0wo2bl30v45.pdf?ysclid=lrrjqn28q884580008.

The disadvantage of this solution is the absence in the panel of tools for identifying polymorphisms of the COMT, ZKSCAN5, SULT2A1, BMF, BCL2L11, CYP2C9 genes.

Another analogue is the panel “Genetic polymorphism of receptors and enzymes of sex hormone metabolism in Moscow”, which includes the following markers: AR, COMT, CYP19A1, CYP27B1, ESR1, ESR2, FADS2, PGR, PPARG, SHBG, SRD5A2, VDR found on the Internet 25.01. 2024 https://dnkom.ru/upload/iblock/081/t29m8it31efe8xq0ve2tnml6snsu6cnu.pdf.

The disadvantage of this solution is the absence in the panel of tools for identifying polymorphisms of the ZKSCAN5, SULT2A1, BMF, BCL2L11, CYP2C9 genes, which in the study results will not provide a full assessment of the risks of changes in the concentration of DHEA-c, one of the most important androgens synthesized by the adrenal cortex [Rozhivanov P .V., Vaks V.V. Dehydroepiandrosterone: physiological role and possibilities of use as a medication. Problems of Endocrinology. 2005;51(2):46-51].

A technical challenge is the difficulty in selecting a combination of genes/polymorphisms/genotypes suitable for determining predisposition to a particular type of steroid hormone metabolism, as evidenced by the almost complete absence on the market of diagnostic panels of polymorphisms (SNPs) suitable for diagnosing disorders of steroid hormone metabolism.

Disclosure of the invention

The technical result of the present invention is to create a panel of polymorphisms and use it to determine the predisposition to the development of disorders of steroid hormone metabolism based on a unique combination of polymorphisms, which allows one to simultaneously identify the most common polymorphisms, makes it possible to conduct a more complete diagnostic coverage and not miss possible changes in the genotype . This panel will also allow you to preventively adjust the patient’s diet and lifestyle, and select an effective dosage for HRT (hormone replacement therapy).

Achievement of the specified technical result is ensured by the proposed panel for determining the predisposition to the development of metabolic disorders of steroid hormones based on the use of a unique combination of genotypes of polymorphisms in the genes of estradiol receptors (ESR1,2), progesterone (PGR), androgen receptor (AR), in the genes of steroid transporter proteins hormones, as well as in the genes for the enzymes catechol-O-methyltransferase (COMT), sulfotransferase 2A1, and cytochrome CYP2C9 genes.

The proposed panel includes tools for detecting the following polymorphisms in a patient sample:

Polymorphic variants of estradiol receptors (ESR1,2), progesterone receptors (PGR), androgen receptor (AR) and catechol-O-methyltransferase enzyme (COMT):

1. ESR1 gene polymorphism NC_000006.12:g.151842200T>C rs2234693

2. ESR1 gene polymorphism NC_000006.12:g.151797984T>G rs2881766

3. ESR1 gene polymorphism NC_000006.12:g.152098960G>A rs2228480

4. ESR2 gene polymorphism NC_000014.9:g.64233098C>T rs4986938

5. ESR2 gene polymorphism NC_000014.9:g.64296935A>G rs2987983

6. PGR gene polymorphism NC_000011.10:g.101062681C>A rs1042838

7. PGR gene polymorphism NC_000014.9:g.64296935A>G rs10895068

8. PGR gene polymorphism NC_000011.10:g.101105243C>G rs590688

9. PGR gene polymorphism NC_000011.10:g.101063100G>T rs1824128

10. PGR gene polymorphism NC_000011.10:g.101039579A>T rs10895054

11. PGR gene polymorphism NC_000011.10:g.101051471G>A rs1042839

12. Trinucleotide repeat length polymorphism (CAG)n in the AR gene, rs3032358

13. AR gene polymorphism NC_000023.11:g.67545785G>A rs6152

14. AR gene polymorphism NC_000023.11:g.67568383G>A rs1204038

15. AR gene polymorphism NC_000023.11:g.67764173G>A rs1337082

16. COMT gene polymorphism NC_000022.11:g.19963748G>A (p.Val158Met) rs4680.

Genetic polymorphic variants responsible for the level of testosterone and DHEA-c in the blood:

17. Polymorphism of the SHBG gene (sex hormone binding globulin) NM_001289114.1:c.-62+4486G>Trs12150660

18. Polymorphism of the SHBG gene (sex hormone binding globulin) NC_000017.11:g.7631360C>T rs6258

19. Polymorphism of the intergenic region NC(000023.11):g.8945785T>C rs5934505

20. Polymorphism of the SHBG gene (sex hormone binding globulin) NM_001289113.1:c.-63-311G>Ars1799941

21. Polymorphism of the ZKSCAN5 gene (zinc finger protein with KRAB and SCAN 5 domains) NM_145102.4:c.772+874C>T, rs11761528

22. Polymorphism of the SULT2A1 gene (sulfotransferase 2A1) g. 48401893G>Ars2637125

23. Polymorphism of the BMF gene (Bcl-2-modifying factor) g. 40068540 A>Grs7181230

24. Polymorphism of the BCL2L11 gene (Bcl-2-like protein)chr2:111191750 G>C rs6738028

25. Polymorphism of the CYP2C9 gene (cytochrome P450 family 2C9) g. 94991513 T>Crs2185570.

The proposed panel can be used to determine susceptibility to disorders of steroid hormone metabolism.

Achievement of the specified technical result is also ensured by the use of a panel of polymorphisms to determine the predisposition to the development of disorders of steroid hormone metabolism based on the use of a unique combination of genotypes of polymorphisms in the genes of estradiol receptors (ESR1,2), progesterone (PGR), androgen receptor (AR), in protein genes - steroid hormone transporters, as well as in the genes of the enzymes catechol-O-methyltransferase (COMT), sulfotransferase 2A1, and cytochrome CYP2C9 genes.

The panel proposed for use includes tools for detecting the following polymorphisms in a patient sample:

Polymorphic variants of estradiol receptors (ESR1,2), progesterone receptors (PGR), androgen receptor (AR) and catechol-O-methyltransferase enzyme (COMT):

1. Polymorphism of the ESR1 gene NC_000006.12:g.151842200T>C rs2234693.

2. ESR1 gene polymorphism NC_000006.12:g.151797984T>G rs2881766

3. ESR1 gene polymorphism NC_000006.12:g.152098960G>A rs2228480

4. Polymorphism of the ESR2 gene NC_000014.9:g.64233098C>T rs4986938.

5. ESR2 gene polymorphism NC_000014.9:g.64296935A>G rs2987983

6. PGR gene polymorphism NC_000011.10:g.101062681C>A rs1042838.

7. PGR gene polymorphism NC_000014.9:g.64296935A>G rs10895068

8. PGR gene polymorphism NC_000011.10:g.101105243C>G rs590688

9. PGR gene polymorphism NC_000011.10:g.101063100G>T rs1824128

10. PGR gene polymorphism NC_000011.10:g.101039579A>T rs10895054

11. PGR gene polymorphism NC_000011.10:g.101051471G>A rs1042839

12. Trinucleotide repeat length polymorphism (CAG)n in the AR gene, rs3032358.

13. AR gene polymorphism NC_000023.11:g.67545785G>A rs6152

14. AR gene polymorphism NC_000023.11:g.67568383G>A rs1204038

15. AR gene polymorphism NC_000023.11:g.67764173G>A rs1337082

16. COMT gene polymorphism NC_000022.11:g.19963748G>A (p.Val158Met) rs4680.

Genetic polymorphic variants responsible for the level of testosterone and DHEA-c in the blood:

17. Polymorphism of the SHBG gene (sex hormone binding globulin) NM_001289114.1:c.-62+4486G>Trs12150660

18. Polymorphism of the SHBG gene (sex hormone binding globulin) NC_000017.11:g.7631360C>T rs6258

19. Polymorphism of the intergenic region NC(000023.11):g.8945785T>C rs5934505

20. Polymorphism of the SHBG gene (sex hormone binding globulin) NM_001289113.1:c.-63-311G>Ars1799941

21. Polymorphism of the ZKSCAN5 gene (zinc finger protein with KRAB and SCAN 5 domains) NM_145102.4:c.772+874C>T,rs11761528

22. Polymorphism of the SULT2A1 gene (sulfotransferase 2A1) g. 48401893G>Ars2637125

23. Polymorphism of the BMF gene (Bcl-2-modifying factor) g. 40068540 A>Grs7181230

24. Polymorphism of the BCL2L11 gene (Bcl-2-like protein) chr2:111191750 G>C rs6738028

25. Polymorphism of the CYP2C9 gene (cytochrome P450 family 2C9) g. 94991513 T>Crs2185570.

Achieving the specified technical result is also ensured by the proposed method for determining the predisposition to the development of metabolic disorders of steroid hormones based on the use of a unique combination of genotypes of polymorphisms in the genes of estradiol receptors (ESR1,2), progesterone (PGR), androgen receptor (AR), in the genes of steroid transporter proteins hormones, as well as in the genes of the enzymes catechol-O-methyltransferase (COMT), sulfotransferase 2A1, and cytochrome CYP2C9 genes.

The proposed method is characterized by the use of means for detecting gene polymorphisms associated with disorders of steroid hormone metabolism in a sample from a patient, and identifying, based on the results of detection of polymorphisms, the patient's possible predisposition to the development of disorders of steroid hormone metabolism.

In this case, the proposed method involves the use of the specified panel of polymorphisms, namely, the method includes identifying the following polymorphisms in a sample from a patient:

Polymorphic variants of estradiol receptors (ESR1,2), progesterone receptors (PGR), androgen receptor (AR) and catechol-O-methyltransferase enzyme (COMT):

1. ESR1 gene polymorphism NC_000006.12:g.151842200T>C rs2234693

2. ESR1 gene polymorphism NC_000006.12:g.151797984T>G rs2881766

3. ESR1 gene polymorphism NC_000006.12:g.152098960G>A rs2228480

4. Polymorphism of the ESR2 gene NC_000014.9:g.64233098C>T rs4986938.

5. ESR2 gene polymorphism NC_000014.9:g.64296935A>G rs2987983

6. PGR gene polymorphism NC_000011.10:g.101062681C>A rs1042838

7. PGR gene polymorphism NC_000014.9:g.64296935A>G rs10895068

8. PGR gene polymorphism NC_000011.10:g.101105243C>G rs590688

9. PGR gene polymorphism NC_000011.10:g.101063100G>T rs1824128

10. PGR gene polymorphism NC_000011.10:g.101039579A>T rs10895054

11. PGR gene polymorphism NC_000011.10:g.101051471G>A rs1042839

12. Trinucleotide repeat length polymorphism (CAG)n in the AR gene, rs3032358

13. AR gene polymorphism NC_000023.11:g.67545785G>A rs6152

14. AR gene polymorphism NC_000023.11:g.67568383G>A rs1204038

15. AR gene polymorphism NC_000023.11:g.67764173G>A rs1337082

16. COMT gene polymorphism NC_000022.11:g.19963748G>A (p.Val158Met) rs4680.

Genetic polymorphic variants responsible for the level of testosterone and DHEA-c in the blood:

17. Polymorphism of the SHBG gene (sex hormone binding globulin) NM_001289114.1:c.-62+4486G>Trs12150660

18. Polymorphism of the SHBG gene (sex hormone binding globulin) NC_000017.11:g.7631360C>T rs6258

19. Polymorphism of the intergenic region NC(000023.11):g.8945785T>C rs5934505

20. Polymorphism of the SHBG gene (sex hormone binding globulin) NM_001289113.1:c.-63-311G>Ars179994

21. Polymorphism of the ZKSCAN5 gene (zinc finger protein with KRAB and SCAN 5 domains) NM_145102.4:c.772+874C>T, rs11761528

22. Polymorphism of the SULT2A1 gene (sulfotransferase 2A1) g. 48401893G>Ars2637125

23. Polymorphism of the BMF gene (Bcl-2-modifying factor) g. 40068540 A>Grs7181230

24. Polymorphism of the BCL2L11 gene (Bcl-2-like protein chr2: 111191750 G>C rs6738028

25. Polymorphism of the CYP2C9 gene (cytochrome P450 family 2C9) g. 94991513T>Crs2185570.

The presence of at least one of these polymorphisms in a patient indicates a possible predisposition to the development of disorders of steroid hormone metabolism.

The achievement of the specified technical result is also ensured by the use of the panel in the specified method for determining the predisposition to the development of disorders of the metabolism of steroid hormones, where the application is characterized by the implementation of the specified stages of the method, namely the use of means for detecting gene polymorphisms in a sample from the patient associated with disorders of the metabolism of steroid hormones, and identifying based on the results of detecting polymorphisms of a patient’s possible predisposition to the development of disorders of steroid hormone metabolism, where the method includes identifying the following polymorphisms in a sample from the patient:

Polymorphic variants of estradiol receptors (ESR1,2), progesterone receptors (PGR), androgen receptor (AR) and catechol-O-methyltransferase enzyme (COMT):

1. ESR1 gene polymorphism NC_000006.12:g.151842200T>C rs2234693

2. ESR1 gene polymorphism NC_000006.12:g.151797984T>G rs2881766

3. ESR1 gene polymorphism NC_000006.12:g.152098960G>A rs2228480

4. ESR2 gene polymorphism NC_000014.9:g.64233098C>T rs4986938

5. ESR2 gene polymorphism NC_000014.9:g.64296935A>G rs2987983

6. PGR gene polymorphism NC_000011.10:g.101062681C>A rs1042838

7. PGR gene polymorphism NC_000014.9:g.64296935A>G rs10895068

8. PGR gene polymorphism NC_000011.10:g.101105243C>G rs590688

9. PGR gene polymorphism NC_000011.10:g.101063100G>T rs1824128

10. PGR gene polymorphism NC_000011.10:g.101039579A>T rs10895054

11. PGR gene polymorphism NC_000011.10:g.101051471G>A rs1042839

12. Trinucleotide repeat length polymorphism (CAG)n in the AR gene, rs3032358

13. AR gene polymorphism NC_000023.11:g.67545785G>A rs6152

14. AR gene polymorphism NC_000023.11:g.67568383G>A rs1204038

15. AR gene polymorphism NC_000023.11:g.67764173G>A rs1337082

16. COMT gene polymorphism NC_000022.11:g.19963748G>A (p.Val158Met) rs4680.

Genetic polymorphic variants responsible for the level of testosterone and DHEA-c in the blood:

17. Polymorphism of the SHBG gene (sex hormone binding globulin) NM_001289114.1:c.-62+4486G>Trs12150660

18. Polymorphism of the SHBG gene (sex hormone binding globulin) NC_000017.11:g.7631360C>T rs6258

19. Polymorphism of the intergenic region NC(000023.11):g.8945785T>C rs5934505

20. Polymorphism of the SHBG gene (sex hormone binding globulin) NM_001289113.1:c.-63-311G>Ars1799941

21. Polymorphism of the ZKSCAN5 gene (zinc finger protein with KRAB and SCAN 5 domains) NM_145102.4:c.772+874C>T, rs11761528

22. Polymorphism of the SULT2A1 gene (sulfotransferase 2A1) g. 48401893G>Ars2637125

23. Polymorphism of the BMF gene (Bcl-2-modifying factor) g. 40068540 A>Grs7181230

24. Polymorphism of the BCL2L11 gene (Bcl-2-like protein) chr2:111191750 G>C rs6738028

25. Polymorphism of the CYP2C9 gene (cytochrome P450 family 2C9) g. 94991513 T>Crs2185570.

Polymorphism of the CYP2C9 gene (cytochrome P450 family 2C9) g. 94991513 T>Crs2185570, where the presence of at least one of these polymorphisms in the patient indicates a possible predisposition to the development of disorders of steroid hormone metabolism.

The proposed invention allows us to make an assumption about the type of steroidogenesis and the possible development of metabolic disorders of steroid hormones.

To find a combination of polymorphisms suitable for creating a panel that would make it possible to make an unambiguous assumption about the type of steroidogenesis, a study was conducted taking into account the following factors indicated in Table 1.

Table 1

Genes and polymorphic variants Pathological genotype Possible effects ZKSCAN5 (Zinc Finger Protein With KRAB And SCAN Domains) (NM_145102.4):c.772+874C>T, rs11761528 Homozygote - T/T, heterozygote C/T Carriage of the T allele is associated with a decrease in the concentration of DHEAs in the blood. SULT2A1 chr19 48401893 G>A rs2637125 Homozygote - A/A heterozygote G/A Carriage of the A allele is associated with a decrease in the concentration of DHEAs in the blood. BMF chr15 40068540 A>G rs7181230 Homozygote - G/G heterozygote G/A Carriage of the G allele is associated with an increase in the concentration of DHEAs in the blood. BCL2L11 chr2:111191750 G>C rs6738028 Homozygote - G/G heterozygote G/C Carriage of the G allele is associated with a decrease in the concentration of DHEAs in the blood CYP2C9 chr10:94991513 T>C rs2185570 Homozygote - C/C heterozygote T/C Carriage of the C allele is associated with a decrease in the concentration of DHEAs in the blood SHBG (NM_001289114.1):c.-62+4486G>T rs12150660 Homozygote - T/T Carriage of the T allele is associated with an increase in serum total testosterone and SHBG SHBG(NM_001289113.1):c.-63-311G>A rs1799941 Homozygote - A/A heterozygote G/A Carriage of allele A - high SHBG, increased total testosterone, decreased free testosterone NC_000017.11:g.7631360C>T rs6258 Homozygote - T/T Carriage of the T allele is associated with a decrease in total testosterone and SHBG in serum **** NC_000023.11:g.8945785T>C rs5934505 Homozygote - T/T Carriage of the T allele is associated with a decrease in total testosterone. ESR1 NC_000006.12:g.151842200T>C rs2234693 Homozygote S/C heterozygote S/T Carriage of the C allele is associated with the risk of developing breast cancer and endomitriosis NC_000006.12:g.152098960G>A rs2228480 Homozygote - A/A heterozygote G/A Carriage of the A allele is associated with a reduced risk of breast cancer and increases the risk of migraine. NC_000006.12:g.151797984T>G rs2881766 Homozygote - T/T Carriage of the T allele is associated with an increased risk of breast cancer, hypertension in pregnant women, and the development of thyroid tumors ESR2 NC_000014.9:g.64233098C>T rs4986938 Homozygote - T/T heterozygote - C/T Carriage of the T allele is associated with endometrial hyperplasia, obesity, metabolic syndrome, and Graves' disease. NC_000014.9:g.64296935A>G rs2987983 Homozygote - G/G heterozygote G/A Carriage of the G allele is associated with an increased risk of developing prostate and breast cancer PGR NC_000011.10:g.101062681C>A rs1042838 Homozygote - A/A heterozygote - C/A Carriage of the A allele is associated with the risk of developing endometrial ovarian and breast cancer NC_000014.9:g.64296935A>G rs10895068 Homozygote - G/G heterozygote - G/A Carriage of the G allele is associated with the risk of developing breast cancer NC_000011.10:g.101105243C>G rs590688 Homozygote - G/G Carriage of the G allele is associated with the risk of developing breast cancer NC_000011.10:g.101051471G>A rs1042839 Homozygote - A/A heterozygote G/A Carriage of the A allele is associated with the risk of developing ovarian endometrial cancer NC_000011.10:g.101039579A>T rs10895054 Homozygote - T/T Carriage of the T allele is associated with the risk of developing breast cancer AR VNTR (CAG)n rs3032358 (CAG)n<18 (CAG)n>24 (CAG)n < 18 is associated with: Increased sensitivity to androgens. Increased risk of developing and the likelihood of recurrence of prostate cancer, more aggressive tumor development. Polycystic ovary syndrome. Increased risk of developing breast and endometrial cancer. Hyperandrogenism in women. (CAG)n >24 is associated with: Decreased sensitivity to androgens. Impaired spermatogenesis. Decreased fertility. Hypogonadism. Pathology of pregnancy. Rheumatoid arthritis. Baldness. More than 40 - spinobulbar muscular atrophy (Kennedy disease). NC_000023.11:g.67545785G>A rs6152 Homozygote - G/G heterozygote G/A Carriage of allele A - reduces the risk of baldness, increases the risk of endometrial cancer, increases DHEA, free and total testosterone in plasma. NC_000023.11:g.67568383G>A rs1204038 Homozygote - A/A heterozygote G/A Carriage of allele A increases the risk of developing endometrial cancer, increasing the level of free testosterone in plasma. NC_000023.11:g.67764173G>A rs1337082 Homozygote G/G Heterozygote G/A Carriage of the G allele increases the risk of developing endometrial cancer COMT NC_000022.11:g.19963748G>A (p.Val158Met) rs4680 Absent The G/G genotype is associated with - high enzyme activity, low dopamine levels, high pain threshold, better resistance to stress, although with a moderate decrease in research performance. With the A/A genotype, there is a 3- and 4-fold decrease in enzymatic activity compared to the G/G genotype. As a result, there is more dopamine in the prefrontal cortex, which may be responsible for many neuropsychological associations - lower pain thresholds, increased vulnerability to stress, but also more efficient information processing in most conditions and greater analytical abilities. Decreased enzyme activity contributes to physiologically elevated estradiol levels, which may be associated with an increased risk of breast cancer.

According to the results of the studies, eleven genes were selected from a statistically significant cohort of patients, namely ZKSCAN5, SULT2A1, BMF, BCL2L11, CYP2C9, SHBG, ESR1, ESR2, PGR, AR and COMT, certain polymorphisms of these genes were identified and those most associated with disorders were identified steroidogenesis genotypes of the specified polymorphisms, where the patient has at least one of the following polymorphisms:

Polymorphic variants of estradiol receptors (ESR1,2), progesterone receptors (PGR), androgen receptor (AR) and catechol-O-methyltransferase enzyme (COMT):

1. ESR1 gene polymorphism NC_000006.12:g.151842200T>C rs2234693

2. ESR1 gene polymorphism NC_000006.12:g.151797984T>G rs2881766

3. ESR1 gene polymorphism NC_000006.12:g.152098960G>A rs2228480

4. ESR2 gene polymorphism NC_000014.9:g.64233098C>T rs4986938

5. ESR2 gene polymorphism NC_000014.9:g.64296935A>G rs2987983

6. PGR gene polymorphism NC_000011.10:g.101062681C>A rs1042838

7. PGR gene polymorphism NC_000014.9:g.64296935A>G rs10895068

8. PGR gene polymorphism NC_000011.10:g.101105243C>G rs590688

9. PGR gene polymorphism NC_000011.10:g.101063100G>T rs1824128

10. PGR gene polymorphism NC_000011.10:g.101039579A>T rs10895054

11. PGR gene polymorphism NC_000011.10:g.101051471G>A rs1042839

12. Trinucleotide repeat length polymorphism (CAG)n in the AR gene, rs3032358

13. AR gene polymorphism NC_000023.11:g.67545785G>A rs6152

14. AR gene polymorphism NC_000023.11:g.67568383G>A rs1204038

15. AR gene polymorphism NC_000023.11:g.67764173G>A rs1337082

16. COMT gene polymorphism NC_000022.11:g.19963748G>A (p.Val158Met) rs4680.

Genetic polymorphic variants responsible for the level of testosterone and DHEA-c in the blood:

17. Polymorphism of the SHBG gene (sex hormone binding globulin) NM_001289114.1:c.-62+4486G>Trs12150660

18. Polymorphism of the SHBG gene (sex hormone binding globulin) NC_000017.11:g.7631360C>T rs6258

19. Polymorphism of the intergenic region NC(000023.11):g.8945785T>C rs5934505

20. Polymorphism of the SHBG gene (sex hormone binding globulin) NM_001289113.1:c.-63-311G>Ars1799941

21. Polymorphism of the ZKSCAN5 gene (zinc finger protein with KRAB and SCAN 5 domains) NM_145102.4:c.772+874C>T, rs11761528

22. Polymorphism of the SULT2A1 gene (sulfotransferase 2A1) g. 48401893G>Ars2637125

23. Polymorphism of the BMF gene (Bcl-2-modifying factor) g. 40068540 A>Grs7181230

24. Polymorphism of the BCL2L11 gene (Bcl-2-like protein) chr2:111191750 G>C rs6738028

25. Polymorphism of the CYP2C9 gene (cytochrome P450 family 2C9) g. 94991513 T>Crs2185570

Determining exactly this combination of genotypes of polymorphisms of these genes is diagnostically significant, since it covers a sufficient number of genes involved in the formation and transport of steroid hormones, the sensitivity of steroid receptors and the metabolism of steroid hormones, which makes it possible to identify the risk at all stages of steroidogenesis.

Implementation of the invention.

The proposed invention is based on a molecular genetic study of samples of cells, tissues or physiological fluids from patients. As a non-limiting example, a sample of peripheral blood from patients aged 18 years or older was examined for the following groups of mutations in the genes ZKSCAN5, SULT2A1, BMF, BCL2L11, CYP2C9, SHBG, ESR1, ESR2, PGR, AR and COMT. The molecular genetic research procedure is carried out as follows:

For genotyping, DNA isolated from human blood cells is used. For isolation, for example, the “PROBA-GS-GENETIKA” kit (DNA technology, Russia) or another set of reagents is used that allows you to isolate DNA from peripheral blood with a concentration above 5 ng/μl. The isolation procedure follows the manufacturer's protocol.

As reagents for real-time polymerase chain reaction (RT-PCR), for example, the “5X ScreenMix-HS” kit (Evrogen, Russia) is used. In the polymerase chain reaction, probes such as TaqMan are used. The selection of oligonucleotides (primers and probes) was carried out using, for example, PrimerExpress v3 software (Thermo, USA). The synthesis of oligonucleotides is carried out, for example, on an AppliedBiosistems 3900 device (Thermo, USA) by order from Lumiprob Rus LLC (Russia).

To carry out PCR, for example, a CFX96 c1000 Touch amplifier (BioRad) with real-time detection of amplification products is used.

Diagnosis is carried out based on data obtained from the results of molecular genetic analysis of gene sections for the presence of risk alleles (Table 2).

Table 2.

Gene Polymorphism Condition associated with risk ZKSCAN5 (Zinc Finger Protein With KRAB And SCAN Domains) (NM_145102.4):c.772+874C>T, rs11761528 Homozygote - T/T, heterozygote C/T SULT2A1 chr19 48401893 G>A rs2637125 Homozygote - A/A heterozygote G/A BMF chr15 40068540 A>G rs7181230 Homozygote - G/G heterozygote G/A BCL2L11 chr2:111191750 G>C rs6738028 Homozygote - G/G heterozygote G/C CYP2C9 chr10:94991513 T>C rs2185570 Homozygote - C/C heterozygote T/C SHBG (NM_001289114.1):c.-62+4486G>T rs12150660 Homozygote - T/T SHBG(NM_001289113.1):c.-63-311G>A rs1799941 Homozygote - A/A heterozygote G/A NC_000017.11:g.7631360C>T rs6258 Homozygote - T/T **** NC_000023.11:g.8945785T>C rs5934505 Homozygote - T/T ESR1 NC_000006.12:g.151842200T>C rs2234693 Homozygote S/C heterozygote S/T NC_000006.12:g.152098960G>A rs2228480 Homozygote - A/A heterozygote G/A NC_000006.12:g.151797984T>G rs2881766 Homozygote - T/T ESR2 NC_000014.9:g.64233098C>T rs4986938 Homozygote - T/T heterozygote - C/T NC_000014.9:g.64296935A>G rs2987983 Homozygote - G/G heterozygote G/A PGR NC_000011.10:g.101062681C>A rs1042838 Homozygote - A/A heterozygote - C/A NC_000014.9:g.64296935A>G rs10895068 Homozygote - G/G heterozygote - G/A NC_000011.10:g.101105243C>G rs590688 Homozygote - G/G NC_000011.10:g.101051471G>A rs1042839 Homozygote - A/A heterozygote G/A NC_000011.10:g.101039579A>T rs10895054 Homozygote - T/T AR VNTR (CAG)n rs3032358 (CAG)n<18 (CAG)n>24 NC_000023.11:g.67545785G>A rs6152 Homozygote - G/G heterozygote G/A NC_000023.11:g.67568383G>A rs1204038 Homozygote - A/A heterozygote G/A NC_000023.11:g.67764173G>A rs1337082 Homozygote G/G Heterozygote G/A COMT NC_000022.11:g.19963748G>A (p.Val158Met) rs4680 Absent

Below are examples demonstrating the possibility of achieving the specified technical result, which show the results of studies of samples from patients and confirmation of the possibility of predicting differences in the formation and transport of steroid hormones, the sensitivity of steroid receptors and the metabolism of steroid hormones, which were confirmed by additional clinical and laboratory studies.

Examples of implementation of the invention

EXAMPLE 1: Patient, born in 1968, male, results of analysis for genetic risk factors for the development of steroid hormone disorders (Table 3).

Table 3.

Index Genotype SHBG gene (sex hormone binding globulin), polymorphism SHBG(NM_001289114.1):c.-62+4486G>T rs12150660 Genotype G/G. Genotype is not associated with changes in SHBG and total testosterone concentrations. SHBG gene (sex hormone binding globulin), polymorphism SHBG(NM_001289113.1):c.-63-311G>A rs1799941 Genotype G/G. Genotype is not associated with changes in SHBG and total testosterone concentrations. SHBG gene (sex hormone binding globulin), polymorphism NC_000017.11:g.7631360C>T rs6258 Genotype C/S. Genotype is not associated with changes in SHBG and total testosterone concentrations. Polymorphism NC_000023.11: g.8945785T>C rs5934505 Genotype T/T. This genotype is associated with a decrease in the concentration of free and total testosterone. Gene ZKSCAN5, polymorphism NM_145102.4:c.772+874C>T, rs11761528 Genotype C/S. The genotype is not associated with changes in the concentration of DHEA-c in the blood. SULT2A1 gene, polymorphism NC_000019.10:g.47898636G>A, rs2637125 Genotype G/A. Carriage of the A allele is associated with a decrease in the concentration of DHEAs in the blood. BMF gene, polymorphism NC_000015.10:g.40068540A>G, rs7181230 Genotype G/A. Carriage of the G allele is associated with an increase in the concentration of DHEAs in the blood. BCL2L11 gene, polymorphism NC_000002.12:g.111191750G>C, rs6738028 Genotype G/G. Carriage of the G allele is associated with a decrease in the concentration of DHEAs in the blood. CYP2C9 gene, polymorphism NC_000010.11:g.94991513T>C, rs2185570 Genotype T/T. The genotype is not associated with changes in the concentration of DHEAs in the blood. Androgen receptor gene mutation ((CAG)nAR) The number of CAG repeats is within normal limits (from 18 to 24) AR gene (androgen receptor) polymorphism rs6152 NC_000023.11:g.67545785G>A Genotype G/G. The G allele is associated with an increased risk of baldness. AR gene (androgen receptor) polymorphism rs1204038 NC_000023.11:g.67568383G>A 77/23 Genotype G/G. There are no associations with pathological conditions. ESR1 gene (estradiol receptor 1), polymorphism NC_000006.12:g.151842200T>C rs2234693 Genotype T/T. There are no associations with pathological conditions. ESR1 gene (estradiol receptor type 1) polymorphism rs2881766 NC_000006.12:g.151797984T>G 78/22 Genotype T/G - the G allele is associated with an increased risk of thyroid tumors. ESR2 gene (estradiol receptor 2), polymorphism NC_000014.9:g.64233098C>T rs4986938 Genotype C/S. There are no associations with pathological conditions. ESR2 gene (estradiol receptor type 2) polymorphism rs2987983 NC_000014.9:g.64296935A>G 65/35 Genotype A/A, no associations with pathological conditions. PGR gene (progesterone receptor), polymorphism NC_000014.9:g.64233098C>T rs1042838 Genotype C/S. There are no associations with pathological conditions. PGR gene (progesterone receptor) polymorphism rs10895068 (+331G/A) NC_000014.9:g.64296935A>G 65/35 The A/G genotype is associated with a 1.5 increase in the risk of breast cancer in women; in men, associations with pathological conditions are unknown. PGR gene (progesterone receptor) polymorphism rs590688 NC_000011.10:g.101105243C>G 53/47 Genotype C/S. There are no associations with pathological conditions. COMT gene (Catechol-O-methyltransferase), polymorphism NC_000022.11:g.19963748G>A (p.Val158Met) rs4680 Genotype G/A. Not associated with changes in physiological conditions. CYP19A1 gene (aromatase, cytochrome 19A1) polymorphism rs2470152 NC_000015.10:g.51302775G>A Genotype G/G. For the GG genotype, an increase in the level of serum estradiol and estrone in men is shown (up to 13% compared to the AA genotype). The G allele is associated with higher levels of aromatase expression and with increased transformation of androstenedione and testosterone to estrone and estradiol. Does not affect testosterone levels. CYP17A1 gene (17-alpha-hydroxylase) polymorphism rs743572 NC_000010.11:g.102837395A>G Genotype A/G. There are no associations with pathological conditions. SRD5A1 gene (5-alpha reductase 1) polymorphism rs1691053 NC_000005.10:g.6677052T>C Genotype T/T. There are no associations with pathological conditions. SRD5A2 gene (5-alpha reductase 2) polymorphism rs523349 NC_000002.12:g.31580636G>C Genotype G/C. Association of the C allele with the risk of ovarian cancer and recurrence after prostatectomy. Not associated with prostate cancer (reviews and meta-analysis). Comments: The genotype of CAG repeats in the AR gene is 20 (normal variant).

The identified changes in the genotype were confirmed by laboratory data (Table 4).

Table 4.

Testosterone, nmol/l (RR: 13.0-33.00), HPLC/MS method 7.35 Dehydroepiandrosterone (DHEA), nmol/l (RD: 2.0-16.0), HPLC/MS method 3.47

The data obtained indicate that the DHEA value tends to the lower limit of the reference range, the testosterone value is significantly lower than normal, i.e. testosterone synthesis is impaired.

EXAMPLE 2: Patient, born in 1980, female, results of analysis for genetic risk factors for the development of steroid hormone disorders (Table 5).

Table 5.

Index Genotype SHBG gene (sex hormone binding globulin), polymorphism SHBG(NM_001289114.1):c.-62+4486G>T rs12150660 Genotype G/G. Genotype is not associated with changes in SHBG and total testosterone concentrations. SHBG gene (sex hormone binding globulin), polymorphism SHBG(NM_001289113.1):c.-63-311G>A rs1799941 Genotype G/A. For allele A, an increase in the level of SHBG and a decrease in the level of free testosterone are shown. Polymorphism may have a direct effect on SHBG gene transcription. SHBG gene (sex hormone binding globulin), polymorphism NC_000017.11:g.7631360C>T rs6258 Genotype C/S. Genotype is not associated with changes in SHBG and total testosterone concentrations. Polymorphism NC_000023.11: g.8945785T>C rs5934505 Genotype T/T. This genotype is associated with a decrease in the concentration of free and total testosterone. Gene ZKSCAN5, polymorphism NM_145102.4:c.772+874C>T, rs11761528 Genotype C/S. The genotype is not associated with changes in the concentration of DHEAs in the blood. SULT2A1 gene, polymorphism NC_000019.10:g.47898636G>A, rs2637125 Genotype G/A. Carriage of the A allele is associated with a decrease in the concentration of DHEAs in the blood. BMF gene, polymorphism NC_000015.10:g.40068540A>G, rs7181230 Genotype A/A. The genotype is not associated with changes in the concentration of DHEAs in the blood. BCL2L11 gene, polymorphism NC_000002.12:g.111191750G>C, rs6738028 Genotype C/C. The genotype is not associated with changes in the concentration of DHEAs in the blood. CYP2C9 gene, polymorphism NC_000010.11:g.94991513T>C, rs2185570 Genotype T/T. The genotype is not associated with changes in the concentration of DHEAs in the blood. Androgen receptor gene mutation ((CAG)nAR) The number of CAG repeats is within normal limits (from 18 to 24) ESR1 gene (estradiol receptor 1), polymorphism NC_000006.12:g.151842200T>C rs2234693 Genotype T/C. Carriage of the C allele is associated with hypertension, placental dysfunction, preeclampsia, myocardial infarction and ischemic cerebral stroke. ESR2 gene (estradiol receptor 2), polymorphism NC_000014.9:g.64233098C>T rs4986938 Genotype C/S. There are no associations with pathological conditions. PGR gene (progesterone receptor), polymorphism NC_000014.9:g.64233098C>T rs1042838 Genotype C/A. Increased risk of developing ovarian endometrial cancer. COMT gene (Catechol-O-methyltransferase), polymorphism NC_000022.11:g.19963748G>A (p.Val158Met) rs4680 Genotype G/G. The genotype is associated with high COMT enzymatic activity and low dopamine levels, increased pain threshold, high resistance to stress, and is also associated with reduced interest in research and with a moderate decrease in the effectiveness of activities aimed at cognition. Comments on the sample: Genotype of CAG repeats in the AR gene 20/23 (normal variant)

The identified changes in the genotype were confirmed by laboratory data (Table 6).

Table 6.

Testosterone, nmol/l (RR: 0.8-2.70), HPLC/MS method 0.54 Dehydroepiandrosterone (DHEA), nmol/l (RD: 2.0-16.0), HPLC/MS method 3.59

The data obtained indicate that the DHEA value tends to the lower limit of the reference range, the testosterone value is below normal, i.e. testosterone synthesis is impaired.

EXAMPLE 3: Patient, born in 1979, male, results of analysis for genetic risk factors for the development of steroid hormone disorders (Table 7).

Table 7.

Index Genotype SHBG gene (sex hormone binding globulin), polymorphism SHBG(NM_001289114.1):c.-62+4486G>T rs12150660 Genotype G/G. Genotype is not associated with changes in SHBG and total testosterone concentrations. SHBG gene (sex hormone binding globulin), polymorphism SHBG(NM_001289113.1):c.-63-311G>A rs1799941 Genotype G/A. For allele A, an increase in the level of SHBG and a decrease in the level of free testosterone are shown. Polymorphism may have a direct effect on SHBG gene transcription. SHBG gene (sex hormone binding globulin), polymorphism NC_000017.11:g.7631360C>T rs6258 Genotype C/S. Genotype is not associated with changes in SHBG and total testosterone concentrations. Polymorphism NC_000023.11: g.8945785T>C rs5934505 Genotype S/T. The genotype is not associated with changes in the concentration of SHBG and total testosterone (compared to carriers of the C allele). Gene ZKSCAN5, polymorphism NM_145102.4:c.772+874C>T, rs11761528 Genotype S/T. Carriage of the T allele is associated with a decrease in the concentration of DHEAs in the blood. SULT2A1 gene, polymorphism NC_000019.10:g.47898636G>A, rs2637125 Genotype G/G. The genotype is not associated with changes in the concentration of DHEAs in the blood. BMF gene, polymorphism NC_000015.10:g.40068540A>G, rs7181230 Genotype A/A. The genotype is not associated with changes in the concentration of DHEAs in the blood. BCL2L11 gene, polymorphism NC_000002.12:g.111191750G>C, rs6738028 Genotype G/G. Carriage of the G allele is associated with a decrease in the concentration of DHEAs in the blood (compared to carriers of the C allele). CYP2C9 gene, polymorphism NC_000010.11:g.94991513T>C, rs2185570 Genotype T/T. The genotype is not associated with changes in the concentration of DHEAs in the blood. Androgen receptor gene mutation ((CAG)nAR) The number of CAG repeats is within normal limits (from 18 to 24) AR gene (androgen receptor) polymorphism rs6152 NC_000023.11:g.67545785G>A Genotype G/A. The A allele is associated with endometrial cancer, increased DHEA and total testosterone. The G allele is associated with an increased risk of baldness in men. AR gene (androgen receptor) polymorphism rs1204038 NC_000023.11:g.67568383G>A 77/23 Genotype G/G. There are no associations with pathological conditions. ESR1 gene (estradiol receptor 1), polymorphism NC_000006.12:g.151842200T>C rs2234693 Genotype T/T. There are no associations with pathological conditions. ESR1 gene (estradiol receptor type 1) polymorphism rs2881766 NC_000006.12:g.151797984T>G 78/22 Genotype T/T. There are no associations with pathological conditions. ESR2 gene (estradiol receptor 2), polymorphism NC_000014.9:g.64233098C>T rs4986938 Genotype C/S. There are no associations with pathological conditions. ESR2 gene (estradiol receptor type 2) polymorphism rs2987983 NC_000014.9:g.64296935A>G 65/35 Genotype A/A, no associations with pathological conditions. PGR gene (progesterone receptor), polymorphism NC_000014.9:g.64233098C>T rs1042838 Genotype C/S. There are no associations with pathological conditions. PGR gene (progesterone receptor) polymorphism rs10895068 (+331G/A) NC_000014.9:g.64296935A>G 65/35 The A/A genotype is associated with a 2.5 increase in the risk of breast cancer; in men, associations with pathological conditions are unknown. PGR gene (progesterone receptor) polymorphism rs590688 NC_000011.10:g.101105243C>G 53/47 Genotype G/G. There is an association with breast cancer in the presence of risk alleles rs1042838 and rs10895068. COMT gene (Catechol-O-methyltransferase), polymorphism NC_000022.11:g.19963748G>A (p.Val158Met) rs4680 Genotype A/A. Associated with low COMT activity (up to 25% of enzyme activity in carriers of the G/G genotype), increased dopamine in the anterior cerebral cortex, tendency to anxiety, low pain threshold, increased vulnerability to stress, and is also associated with an advantage in performing cognitive tasks, but only in a calm environment. Also, a number of studies show that people with the A/A genotype may have more effective adjustment (not to be confused with resistance) to stress, a higher level of self-control and higher reactivity to positive stimuli. CYP19A1 gene (aromatase, cytochrome 19A1) polymorphism rs2470152 NC_000015.10:g.51302775G>A Genotype G/G. For the GG genotype, an increase in the level of serum estradiol and estrone in men is shown (up to 13% compared to the AA genotype). The G allele is associated with higher levels of aromatase expression and with increased transformation of androstenedione and testosterone to estrone and estradiol. Does not affect testosterone levels. CYP17A1 gene (17-alpha-hydroxylase) polymorphism rs743572 NC_000010.11:g.102837395A>G Genotype G/G. Associated with polycystic ovary syndrome and prostate cancer (in certain populations). SRD5A1 gene (5-alpha reductase 1) polymorphism rs1691053 NC_000005.10:g.6677052T>C Genotype T/T. There are no associations with pathological conditions. SRD5A2 gene (5-alpha reductase 2) polymorphism rs523349 NC_000002.12:g.31580636G>C Genotype G/C. Association of the C allele with the risk of ovarian cancer and recurrence after prostatectomy. Not associated with prostate cancer (reviews and meta-analysis).

The identified changes in the genotype were confirmed by laboratory data (Table 8).

Table 8.

Testosterone, nmol/l (RD: 13.0-33.0), HPLC/MS method 12.8 Dehydroepiandrosterone (DHEA), nmol/l (RD: 2.0-16.0), HPLC/MS method 14.2 Cortisol, nmol/l (RD: 130.0-270.0), HPLC/MS method 421 Estradiol, nmol/l (RR: 0.07-0.2) 0.2 SHBG, nmol/l (RZ: 18-54) 18.3

The data obtained indicate that the testosterone value is below normal, while the estradiol value is at the upper limit of the reference range, and the transformation of testosterone into estradiol is increased. There is also a high level of cortisol, which is most likely due to low COMT activity. The low level of SHBG is noteworthy.

EXAMPLE 4: Patient, born in 1972, female, results of analysis for genetic risk factors for the development of steroid hormone disorders (Table 9).

Table 9.

Index Genotype SHBG gene (sex hormone binding globulin), polymorphism SHBG(NM_001289114.1):c.-62+4486G>T rs12150660 Genotype G/G. Genotype is not associated with changes in SHBG and total testosterone concentrations. SHBG gene (sex hormone binding globulin), polymorphism SHBG(NM_001289113.1):c.-63-311G>A rs1799941 Genotype G/A. For allele A, an increase in the level of SHBG and a decrease in the level of free testosterone are shown. Polymorphism may have a direct effect on SHBG gene transcription. SHBG gene (sex hormone binding globulin), polymorphism NC_000017.11:g.7631360C>T rs6258 Genotype C/S. Genotype is not associated with changes in SHBG and total testosterone concentrations. Polymorphism NC_000023.11: g.8945785T>C rs5934505 Genotype T/T. This genotype is associated with a decrease in the concentration of free and total testosterone. Gene ZKSCAN5, polymorphism NM_145102.4:c.772+874C>T, rs11761528 Genotype C/S. The genotype is not associated with changes in the concentration of DHEAs in the blood. SULT2A1 gene, polymorphism NC_000019.10:g.47898636G>A, rs2637125 Genotype G/G. The genotype is not associated with changes in the concentration of DHEAs in the blood. BMF gene, polymorphism NC_000015.10:g.40068540A>G, rs7181230 Genotype A/A. The genotype is not associated with changes in the concentration of DHEAs in the blood. BCL2L11 gene, polymorphism NC_000002.12:g.111191750G>C, rs6738028 Genotype G/G. Carriage of the G allele is associated with a decrease in the concentration of DHEAs in the blood. CYP2C9 gene, polymorphism NC_000010.11:g.94991513T>C, rs2185570 Genotype T/T. The genotype is not associated with changes in the concentration of DHEAs in the blood. Androgen receptor gene mutation ((CAG)nAR) The number of CAG repeats is within normal limits (from 18 to 24) ESR1 gene (estradiol receptor 1), polymorphism NC_000006.12:g.151842200T>C rs2234693 Genotype T/C. Carriage of the C allele is associated with hypertension, placental dysfunction, preeclampsia, and myocardial infarction. ESR2 gene (estradiol receptor 2), polymorphism NC_000014.9:g.64233098C>T rs4986938 Genotype C/S. There are no associations with pathological conditions. PGR gene (progesterone receptor), polymorphism NC_000014.9:g.64233098C>T rs1042838 Genotype C/S. There are no associations with pathological conditions. COMT gene (Catechol-O-methyltransferase), polymorphism NC_000022.11:g.19963748G>A (p.Val158Met) rs4680 Genotype G/A. Not associated with changes in physiological conditions. Comments on the sample: genotype of CAG repeats in the AR gene 22/21

The identified changes in the genotype were confirmed by laboratory data (Table 10).

Table 10.

SHBG, nmol/l (RZ: 27-128) 163.0

The data obtained indicate that the SHBG value is significantly higher than normal, and the transcription of the SHBG gene is increased.

EXAMPLE 5: Patient, born in 1974, female, results of analysis for genetic risk factors for the development of steroid hormone disorders (Table 11).

Table 11.

Index Genotype SHBG gene (sex hormone binding globulin), polymorphism SHBG(NM_001289114.1):c.-62+4486G>T rs12150660 Genotype G/T. Carriage of the T allele is associated with increased concentrations of SHBG and total testosterone. SHBG gene (sex hormone binding globulin), polymorphism SHBG(NM_001289113.1):c.-63-311G>A rs1799941 Genotype G/A. For allele A, an increase in the level of SHBG and a decrease in the level of free testosterone are shown. Polymorphism may have a direct effect on SHBG gene transcription. SHBG gene (sex hormone binding globulin), polymorphism NC_000017.11:g.7631360C>T rs6258 Genotype C/S. Genotype is not associated with changes in SHBG and total testosterone concentrations. Polymorphism NC_000023.11: g.8945785T>C rs5934505 Genotype S/T. Genotype is not associated with changes in SHBG and total testosterone concentrations. Gene ZKSCAN5, polymorphism NM_145102.4:c.772+874C>T, rs11761528 Genotype C/S. The genotype is not associated with changes in the concentration of DHEAs in the blood. SULT2A1 gene, polymorphism NC_000019.10:g.47898636G>A, rs2637125 Genotype G/A. Carriage of the A allele is associated with a decrease in the concentration of DHEAs in the blood. BMF gene, polymorphism NC_000015.10:g.40068540A>G, rs7181230 Genotype G/G. Carriage of the G allele is associated with an increase in the concentration of DHEAs in the blood. BCL2L11 gene, polymorphism NC_000002.12:g.111191750G>C, rs6738028 Genotype G/G. Carriage of the G allele is associated with a decrease in the concentration of DHEAs in the blood. CYP2C9 gene, polymorphism NC_000010.11:g.94991513T>C, rs2185570 Genotype T/T. The genotype is not associated with changes in the concentration of DHEAs in the blood. Androgen receptor gene mutation ((CAG)nAR) An increase in CAG repeats (>24) in the heterozygous state was detected. ESR1 gene (estradiol receptor 1), polymorphism NC_000006.12:g.151842200T>C rs2234693 Genotype T/T. There are no associations with pathological conditions. ESR2 gene (estradiol receptor 2), polymorphism NC_000014.9:g.64233098C>T rs4986938 Genotype C/S. There are no associations with pathological conditions. PGR gene (progesterone receptor), polymorphism NC_000014.9:g.64233098C>T rs1042838 Genotype C/S. There are no associations with pathological conditions. COMT gene (Catechol-O-methyltransferase), polymorphism NC_000022.11:g.19963748G>A (p.Val158Met) rs4680 Genotype G/G. The genotype is associated with high COMT enzymatic activity and low dopamine levels, increased pain threshold, high resistance to stress, and is also associated with reduced interest in research and with a moderate decrease in the effectiveness of activities aimed at cognition. Comments on the sample: The genotype of CAG repeats in the AR gene is 23/27.

The identified changes in the genotype were confirmed by laboratory data (Table 12).

Table 12.

Testosterone, nmol/l (RR: 0.8-2.70), HPLC/MS method 2.77 Dihydrotestosterone, nmol/l (RD: 0.1-0.91), HPLC/MS method 1.26

The findings suggest that testosterone metabolism is increased, which may be caused by a decreased response to androgens, which increases the risk of cancer or early ovarian inactivation.

EXAMPLE 6: Patient, born in 1988, male, results of analysis for genetic risk factors for the development of steroid hormone disorders (Table 13).

Table 13.

Index Genotype SHBG gene (sex hormone binding globulin), polymorphism SHBG(NM_001289114.1):c.-62+4486G>T rs12150660 Genotype G/G. Genotype is not associated with changes in SHBG and total testosterone concentrations. SHBG gene (sex hormone binding globulin), polymorphism SHBG(NM_001289113.1):c.-63-311G>A rs1799941 Genotype G/A. For allele A, an increase in the level of SHBG and a decrease in the level of free testosterone are shown. Polymorphism may have a direct effect on SHBG gene transcription. SHBG gene (sex hormone binding globulin), polymorphism NC_000017.11:g.7631360C>T rs6258 Genotype C/S. Genotype is not associated with changes in SHBG and total testosterone concentrations. Polymorphism NC_000023.11: g.8945785T>C rs5934505 Genotype S/T. The genotype is not associated with changes in the concentration of SHBG and total testosterone (compared to carriers of the C allele). Gene ZKSCAN5, polymorphism NM_145102.4:c.772+874C>T, rs11761528 Genotype S/T. Carriage of the T allele is associated with a decrease in the concentration of DHEAs in the blood. SULT2A1 gene, polymorphism NC_000019.10:g.47898636G>A, rs2637125 Genotype G/G. The genotype is not associated with changes in the concentration of DHEAs in the blood. BMF gene, polymorphism NC_000015.10:g.40068540A>G, rs7181230 Genotype A/A. The genotype is not associated with changes in the concentration of DHEAs in the blood. BCL2L11 gene, polymorphism NC_000002.12:g.111191750G>C, rs6738028 Genotype G/G. Carriage of the G allele is associated with a decrease in the concentration of DHEAs in the blood (compared to carriers of the C allele). CYP2C9 gene, polymorphism NC_000010.11:g.94991513T>C, rs2185570 Genotype T/T. The genotype is not associated with changes in the concentration of DHEAs in the blood. Androgen receptor gene mutation ((CAG)nAR) The number of CAG repeats is within normal limits (from 18 to 24) AR gene (androgen receptor) polymorphism rs6152 NC_000023.11:g.67545785G>A Genotype G/G. The G allele is associated with an increased risk of baldness in men, but not in women. AR gene (androgen receptor) polymorphism rs1204038 NC_000023.11:g.67568383G>A 77/23 Genotype G/G. There are no associations with pathological conditions. ESR1 gene (estradiol receptor 1), polymorphism NC_000006.12:g.151842200T>C rs2234693 Genotype T/C. Carriage of the C allele is associated with hypertension, placental dysfunction, preeclampsia, myocardial infarction and ischemic cerebral stroke. ESR1 gene (estradiol receptor type 1) polymorphism rs2881766 NC_000006.12:g.151797984T>G 78/22 Genotype T/T. There are no associations with pathological conditions. ESR2 gene (estradiol receptor 2), polymorphism NC_000014.9:g.64233098C>T rs4986938 Genotype C/S. There are no associations with pathological conditions. ESR2 gene (estradiol receptor type 2) polymorphism rs2987983 NC_000014.9:g.64296935A>G 65/35 Genotype A/G, G allele is associated with an increased risk of breast and prostate cancer. PGR gene (progesterone receptor), polymorphism NC_000014.9:g.64233098C>T rs1042838 Genotype C/A. Increased risk of developing ovarian endometrial cancer. PGR gene (progesterone receptor) polymorphism rs10895068 (+331G/A) NC_000014.9:g.64296935A>G 65/35 The A/G genotype is associated with a 1.5 increase in the risk of breast cancer in women; in men, associations with pathological conditions are unknown. PGR gene (progesterone receptor) polymorphism rs590688 NC_000011.10:g.101105243C>G 53/47 Genotype C/G. There is an association with breast cancer in the presence of risk alleles rs1042838 and rs10895068. COMT gene (Catechol-O-methyltransferase), polymorphism NC_000022.11:g.19963748G>A (p.Val158Met) rs4680 Genotype G/A. Not associated with changes in physiological conditions. CYP19A1 gene (aromatase, cytochrome 19A1) polymorphism rs2470152 NC_000015.10:g.51302775G>A Genotype G/G. For the GG genotype, an increase in the level of serum estradiol and estrone in men is shown (up to 13% compared to the AA genotype). The G allele is associated with higher levels of aromatase expression and increased transformation of androstenedione and testosterone to estrone and estradiol. Does not affect testosterone levels. CYP17A1 gene (17-alpha-hydroxylase) polymorphism rs743572 NC_000010.11:g.102837395A>G Genotype G/G. Associated with polycystic ovary syndrome and prostate cancer (in certain populations). SRD5A1 gene (5-alpha reductase 1) polymorphism rs1691053 NC_000005.10:g.6677052T>C Genotype T/T. There are no associations with pathological conditions. SRD5A2 gene (5-alpha reductase 2) polymorphism rs523349 NC_000002.12:g.31580636G>C Genotype G/C. Association of the C allele with the risk of ovarian cancer and recurrence after prostatectomy. Not associated with prostate cancer (reviews and meta-analysis).

The identified changes in the genotype were confirmed by laboratory data (Table 14).

Table 14.

Testosterone, nmol/l (RD: 13.0-33.0), HPLC/MS method 12.1 Dihydrotestosterone, nmol/l (RD: 0.4-2.65), HPLC/MS method <0.29 Estradiol, nmol/l (RR: 0.07-0.2) 0.19

The data obtained indicate that the value of testosterone and dihydrotestosterone is below normal, while the value of estradiol is at the upper limit of the reference range, the transformation of testosterone into estradiol is increased.

EXAMPLE 7: Patient, born in 1967, male, results of analysis for genetic risk factors for the development of steroid hormone disorders (Table 15).

Table 15.

Index Genotype SHBG gene (sex hormone binding globulin), polymorphism SHBG(NM_001289114.1):c.-62+4486G>T rs12150660 Genotype G/G. Genotype is not associated with changes in SHBG and total testosterone concentrations. SHBG gene (sex hormone binding globulin), polymorphism SHBG(NM_001289113.1):c.-63-311G>A rs1799941 Genotype G/A. For allele A, an increase in the level of SHBG and a decrease in the level of free testosterone are shown. Polymorphism may have a direct effect on SHBG gene transcription. SHBG gene (sex hormone binding globulin), polymorphism NC_000017.11:g.7631360C>T rs6258 Genotype C/S. Genotype is not associated with changes in SHBG and total testosterone concentrations. Polymorphism NC_000023.11: g.8945785T>C rs5934505 Genotype S/T. The genotype is not associated with changes in the concentration of SHBG and total testosterone (compared to carriers of the C allele). Gene ZKSCAN5, polymorphism NM_145102.4:c.772+874C>T, rs11761528 Genotype S/T. Carriage of the T allele is associated with a decrease in the concentration of DHEAs in the blood. SULT2A1 gene, polymorphism NC_000019.10:g.47898636G>A, rs2637125 Genotype G/G. The genotype is not associated with changes in the concentration of DHEAs in the blood. BMF gene, polymorphism NC_000015.10:g.40068540A>G, rs7181230 Genotype G/A. Carriage of the G allele is associated with an increase in the concentration of DHEAs in the blood. BCL2L11 gene, polymorphism NC_000002.12:g.111191750G>C, rs6738028 Genotype G/G. Carriage of the G allele is associated with a decrease in the concentration of DHEAs in the blood (compared to carriers of the C allele). CYP2C9 gene, polymorphism NC_000010.11:g.94991513T>C, rs2185570 Genotype T/T. The genotype is not associated with changes in the concentration of DHEAs in the blood. Androgen receptor gene mutation ((CAG)nAR) A decrease in CAG repeats was detected in the homozygous state (<18). Increased sensitivity to androgens. AR gene (androgen receptor) polymorphism rs6152 NC_000023.11:g.67545785G>A Genotype G/G. The G allele is associated with an increased risk of baldness in men, but not in women . AR gene (androgen receptor) polymorphism rs1204038 NC_000023.11:g.67568383G>A 77/23 Genotype G/G. There are no associations with pathological conditions. ESR1 gene (estradiol receptor 1), polymorphism NC_000006.12:g.151842200T>C rs2234693 Genotype T/C. Carriage of the C allele is associated with hypertension, placental dysfunction, preeclampsia, myocardial infarction and ischemic cerebral stroke. ESR1 gene (estradiol receptor type 1) polymorphism rs2881766 NC_000006.12:g.151797984T>G 78/22 Genotype T/G - allele G is associated with a reduced risk of breast cancer, hypertension in pregnant women, and an increased risk of thyroid tumors. ESR2 gene (estradiol receptor 2), polymorphism NC_000014.9:g.64233098C>T rs4986938 Genotype C/S. There are no associations with pathological conditions. ESR2 gene (estradiol receptor type 2) polymorphism rs2987983 NC_000014.9:g.64296935A>G 65/35 Genotype G/G, the G allele is associated with an increased risk of breast cancer and prostate cancer. PGR gene (progesterone receptor), polymorphism NC_000014.9:g.64233098C>T rs1042838 Genotype C/S. There are no associations with pathological conditions. PGR gene (progesterone receptor) polymorphism rs10895068 (+331G/A) NC_000014.9:g.64296935A>G 65/35 Genotype G/G. There are no associations with pathological conditions. PGR gene (progesterone receptor) polymorphism rs590688 NC_000011.10:g.101105243C>G 53/47 Genotype C/G. There is an association with breast cancer in the presence of risk alleles rs1042838 and rs10895068. COMT gene (Catechol-O-methyltransferase), polymorphism NC_000022.11:g.19963748G>A (p.Val158Met) rs4680 Genotype A/A. Associated with low COMT activity (up to 25% of enzyme activity in carriers of the G/G genotype), increased dopamine in the anterior cerebral cortex, tendency to anxiety, low pain threshold, increased vulnerability to stress, and is also associated with an advantage in performing cognitive tasks, but only in a calm environment. Also, a number of studies show that people with the A/A genotype may have more effective adjustment (not to be confused with resistance) to stress, a higher level of self-control and higher reactivity to positive stimuli. CYP19A1 gene (aromatase, cytochrome 19A1) polymorphism rs2470152 NC_000015.10:g.51302775G>A Genotype G/G. For the GG genotype, an increase in the level of serum estradiol and estrone in men is shown (up to 13% compared to the AA genotype). The G allele is associated with higher levels of aromatase expression and increased transformation of androstenedione and testosterone to estrone and estradiol. Does not affect testosterone levels. CYP17A1 gene (17-alpha-hydroxylase) polymorphism rs743572 NC_000010.11:g.102837395A>G Genotype A/G. There are no associations with pathological conditions. SRD5A1 gene (5-alpha reductase 1) polymorphism rs1691053 NC_000005.10:g.6677052T>C Genotype T/T. There are no associations with pathological conditions. SRD5A2 gene (5-alpha reductase 2) polymorphism rs523349 NC_000002.12:g.31580636G>C Genotype G/C. Association of the C allele with the risk of ovarian cancer and recurrence after prostatectomy. Not associated with prostate cancer (reviews and meta-analysis).

The identified changes in the genotype were confirmed by laboratory data (Table 16).

Table 16.

Testosterone, nmol/l (RD: 13.0-33.0), HPLC/MS method 16.85 Dihydrotestosterone, nmol/l (RD: 0.4-2.65), HPLC/MS method <0.29 Dehydroepiandrosterone (DHEA), nmol/l (RD: 0.4-2.65), by HPLC/MS 2.17

The data obtained indicate that the value of testosterone is within the normal range, and dihydrotestosterone is below normal, the concentration of DHEA is at the lower limit of the reference range.

EXAMPLE 8: Patient, born in 1952, male, results of analysis for genetic risk factors for the development of steroid hormone disorders (Table 17).

Table 17.

Index Genotype SHBG gene (sex hormone binding globulin), polymorphism SHBG(NM_001289114.1):c.-62+4486G>T rs12150660 Genotype G/T. Carriage of the T allele is associated with increased concentrations of SHBG and total testosterone. SHBG gene (sex hormone binding globulin), polymorphism SHBG(NM_001289113.1):c.-63-311G>A rs1799941 Genotype G/A. For allele A, an increase in the level of SHBG and a decrease in the level of free testosterone are shown. Polymorphism may have a direct effect on SHBG gene transcription. SHBG gene (sex hormone binding globulin), polymorphism NC_000017.11:g.7631360C>T rs6258 Genotype C/S. Genotype is not associated with changes in SHBG and total testosterone concentrations. Polymorphism NC_000023.11: g.8945785T>C rs5934505 Genotype T/T. This genotype is associated with a decrease in the concentration of free and total testosterone. Gene ZKSCAN5, polymorphism NM_145102.4:c.772+874C>T, rs11761528 Genotype C/S. The genotype is not associated with changes in the concentration of DHEAs in the blood. SULT2A1 gene, polymorphism NC_000019.10:g.47898636G>A, rs2637125 Genotype A/A. Carriage of the A allele is associated with a decrease in the concentration of DHEAs in the blood. BMF gene, polymorphism NC_000015.10:g.40068540A>G, rs7181230 Genotype G/A. Carriage of the G allele is associated with an increase in the concentration of DHEAs in the blood. BCL2L11 gene, polymorphism NC_000002.12:g.111191750G>C, rs6738028 Genotype G/G. Carriage of the G allele is associated with a decrease in the concentration of DHEAs in the blood. CYP2C9 gene, polymorphism NC_000010.11:g.94991513T>C, rs2185570 Genotype T/T. The genotype is not associated with changes in the concentration of DHEAs in the blood. Androgen receptor gene mutation ((CAG)nAR) The number of CAG repeats is within normal limits (from 18 to 24) ESR1 gene (estradiol receptor 1), polymorphism NC_000006.12:g.151842200T>C rs2234693 Genotype T/C. Carriage of the C allele is associated with hypertension, myocardial infarction and ischemic stroke of the brain. ESR2 gene (estradiol receptor 2), polymorphism NC_000014.9:g.64233098C>T rs4986938 Genotype T/T. Carriage of the T allele is associated with obesity, metabolic syndrome, and Graves' disease. PGR gene (progesterone receptor), polymorphism NC_000014.9:g.64233098C>T rs1042838 Genotype C/S. There are no associations with pathological conditions. COMT gene (Catechol-O-methyltransferase), polymorphism NC_000022.11:g.19963748G>A (p.Val158Met) rs4680 Genotype G/A. Not associated with changes in physiological conditions.

The identified changes in the genotype were confirmed by laboratory data (Table 18).

Table 18.

Antibodies to TSH receptors, U/l (RR: <1.75) 1.99 Dehydroepiandrosterone (DHEA), nmol/l (RD: 0.4-2.65), by HPLC/MS 0.17

The data obtained indicate that the Graves' disease marker is above the reference range (AT to TSH receptors), and the DHEA concentration is below the reference range.

Claims (27)

1. A panel to determine susceptibility to the development of disorders of steroid hormone metabolism, including means for detecting the following polymorphisms in a patient sample: 1) ESR1 gene polymorphism NC_000006.12:g.151842200 T>C rs2234693; 2) ESR1 gene polymorphism NC_000006.12:g.151797984 T>G rs2881766; 3) polymorphism of the ESR1 gene NC_000006.12:g.152098960 G>A rs2228480; 4) ESR2 gene polymorphism NC_000014.9:g.64233098 C>T rs4986938; 5) ESR2 gene polymorphism NC_000014.9:g.64296935 A>G rs2987983; 6) polymorphism of the PGR gene NC_000011.10:g.101062681 C>A rs1042838; 7) polymorphism of the PGR gene NC_000014.9:g.64296935 A>G rs10895068; 8) polymorphism of the PGR gene NC_000011.10:g.101105243 C>G rs590688; 9) polymorphism of the PGR gene NC_000011.10:g.101063100 G>T rs1824128; 10) polymorphism of the PGR gene NC_000011.10:g.101039579 A>T rs10895054; 11) polymorphism of the PGR gene NC_000011.10:g.101051471 G>A rs1042839; 12) trinucleotide repeat length polymorphism (CAG)n in the AR gene, rs3032358; 13) AR gene polymorphism NC_000023.11:g.67545785 G>A rs6152; 14) AR gene polymorphism NC_000023.11:g.67568383 G>A rs1204038; 15) AR gene polymorphism NC_000023.11:g.67764173 G>A rs1337082; 16) COMT gene polymorphism NC_000022.11:g.19963748 G>A (p.Val158Met) rs4680; 17) SHBG gene polymorphism NM_001289114.1:c.-62+4486G>Trs12150660; 18) SHBG gene polymorphism NC_000017.11:g.7631360C>T rs6258; 19) polymorphism of the intergenic region NC_000023.11:g.8945785T>C rs5934505; 20) SHBG gene polymorphism NM_001289113.1:c.-63-311G>Ars1799941; 21) polymorphism of the ZKSCAN5 gene NM_145102.4: c.772+874C>T, rs11761528; 22) polymorphism of the SULT2A1 g gene. 48401893G>Ars2637125; 23) polymorphism of the BMF g gene. 40068540 A>Grs7181230; 24) polymorphism of the BCL2L11 gene chr2:111191750 G>C rs6738028; 25) polymorphism of the CYP2C9 g gene. 94991513 T>Crs2185570. 2. A method for determining predisposition to the development of disorders of steroid hormone metabolism, including the use of the panel according to claim 1, where the presence in the patient of at least one of the polymorphisms determined by the panel according to claim 1 indicates a predisposition to the development of disorders of steroid hormone metabolism.