Relationship Between Premature Menopause-related Genetic Polymorphisms and Level of Serum Micronutrients

Relationship Between Premature Menopause-related Genetic Polymorphisms and Level of Serum Micronutrients
Mohammad Reza Mirinezhad,¹ Mohammad Zamiri Bidary,² Alireza Naghipour,³ Tayebeh Hamzehloei,⁴ Alireza Pasdar,⁵ Majid Ghayour-Mobarhan,^6,*
1. Mashhad University of Medical Sciences
2. Mashhad University of Medical Sciences
3. Mashhad University of Medical Sciences
4. Mashhad University of Medical Sciences
5. Mashhad University of Medical Sciences
6. Mashhad University of Medical Sciences
Introduction: Permanent menstrual cessation in women under 40 years old, is called Premature Menopause (PM). The last data indicated that the prevalence of this condition is about 65 percent higher in Iranian women than meta-analyzed estimates of all females around the world that might be related to earlier age at menarche in Iranian girls and lower age of developing ovarian cancer in Iranian females that requires treatments that could lead to PM. Genome-Wide Association Studies (GWAS) and some other epidemiological investigations found that some specific polymorphisms are highly significantly prevalent in PM cases than healthy controls. Moreover, it has been found that micronutrients including vitamin D, calcium, zinc, and copper are associated with PM and could be related to women reproductive tract. In the present study, we have investigated the association between some known PM-related Single Nucleotide Polymorphisms (SNPs) and important micronutrients serum level.
Methods: 117 PM cases along with 183 controls were selected from Mashhad Stroke and Heart Atherosclerotic Disorder (MASHAD) cohort study. PM diagnosis was established by the following criteria: 1) Menstruation cessation for 12 consecutive months, 2) Age under 40, or 3) Follicle-Stimulating Hormone (FSH) elevation (>40 IU/L) for 3 times (repeated at four-week intervals). In order to assess the SNPs of PM cases and comparing them to healthy controls, we used Allele-Specific Oligonucleotide Polymerase Chain Reaction (ASO-PCR) and Tetra Amplification Refractory Mutation System PCR (ARMS-PCR) methods. Serum level of vitamin D (25-hydroxy vitamin D), copper, zinc, calcium, phosphate, and magnesium were measured using standard methods. Finally, the data were analyzed by SPSS, version 24. By Kolmogorov-Smirnov (KSV) test, all variables were checked for normal distribution. Association of SNPs' genotypes and serum level of micronutrients was examined in PM cases and healthy controls, separately by Man-Whitney analysis. Multivariable logistic regression was performed to determine the relationship between two major genotypes of each SNP and micronutrients serum levels' categories, in an adjusted model for age and Body Mass Index (BMI) confounding variables.
Results: Our study found that following SNPs' genotypes are significantly different between cases and controls: rs4806660 (TMEM), rs451417 (MCM8), rs16991615 (MCM8), rs7246479 (BRSK1), rs244715 (ZNF346), and rs10183486 (TLK1). Serum copper, phosphate, and calcium level were considerably different among the genotypes of rs244715, rs451417, and rs4806660 SNPs, respectively in the PM subjects; While in controls, we found that just copper is different meaningfully in level between rs4806660, rs7246479, rs1046089, and rs2303369 SNPs' genotypes. After adjusting the effect size of other genotypes, age, and BMI, we recognized that in PM cases regarding rs4806660 SNP, decreased serum copper concentration was found 78% lower in TC cases than TT ones. Moreover, in assessing rs244715, serum level of copper was significantly elevated in GG genotype PM cases than cases with AA genotype. These significant differences were not found for controls.
Conclusion: It seems that according to previous investigations, SNPs related to PM are associated with hormonal changes, especially sex hormones, and alterations in metabolic factors. Moreover, it was indicated that sex hormone therapy could cause minor changes in copper level; Indeed, primary ovarian insufficiency, which is a known condition that could lead to PM, is associated with serum level of copper. Thus, it might be concluded that PM-related SNPs could cause changes to sex hormones and these hormones might affect the concentration of copper in these cases. In this study, rs4806660 and rs244715 genotypes were found to be related to abnormalities in PM cases' serum copper level.
Keywords: Premature menopause; Micronutrients; Copper; Medical genetics; Polymorphisms