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Hypermethylation and down-regulation of vitamin D receptor (VDR) as contributing factors for polycystic ovary syndrome (PCOS): a case–control study from Kashmir, North India

  • Gynecologic Endocrinology and Reproductive Medicine
  • Published:
Archives of Gynecology and Obstetrics Aims and scope Submit manuscript

Abstract

Background

Polycystic ovarian syndrome (PCOS) is a prevailing endocrinopathy affecting a significant population of women of reproductive age across the globe. A myriad set of complex intertwined factors ranging from etiological, genetic, and epigenetic reasons cause this disorder. Out of the different factors, vitamin D shows an imperative aspect in health and fertility of women with polycystic ovary syndrome (PCOS). The importance of vitamin D is facilitated by vitamin D receptor (VDR), a ligand-dependent transcription factor in the steroid/ thyroid hormone receptor superfamily that controls the pleiotropic biological properties of vitamin D.

Purpose

The purpose of this study was to evaluate the role of promoter methylation of the VDR gene, a transcription factor with numerous biological utilities, with its relative expression and clinico-pathological findings and outcomes.

Methodology

A total of 200 blood samples were collected, 100 from PCOS case subjects, and 100 from the normal healthy controls respectively, which were assessed by qRT-PCR for determining the expression summary. MS-PCR technique was used for analyzing the promoter methylation status of the VDR gene. Blood samples were withdrawn, respectively, for each case and the control study separately experimented for different stages for the given study, of which estimation of vitamin D was also a part.

Results

In this test-versus-control study, first, the promoter methylation status of VDR gene was identified which was found more prominent i.e., hyper-methylation of the VDR gene was identified in 84 cases (84%), and in the normal healthy controls, it was found (62%). The promoter methylation status of the VDR gene has remarkably shown the results with a significant difference (p value < 0.0001*). Second, the expression analysis of VDR gene was found to be strongly downregulated in majority (64%) of PCOS case samples analyzed by means fold change of 0.8743 (± 0.06466) (p value 0.0054**). This result is, therefore, indicative of VDR gene role in PCOS pathogenesis as the said gene is downregulated. Moreover, compared to the vitamin D parameter, hyper-methylation and expression analysis of the VDR promoter gene were found to correspond to some associations with PCOS. Certain case-and-control study analyses showed that patients with normal vitamin D levels showed less indicative effects of PCOS and vice versa.

Conclusion

Our study, being exclusive from Kashmir, one of the foremost specified that VDR confirms anomalous methylation configuration in PCOS with subsequent downregulation in the gene expression i.e., there is an inverse correlation among VDR gene expression (downregulated) and methylation status (hyper-methylated) from the conclusion of our PCOS case-versus-control study.

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Data availability

This work has been published in our earlier artcile titled “Clinical profiling of Polycystic Ovary Syndrome patients in Kashmir Population”. The article details can be checked from: [http://www.matrixscipharma.org IP: 246.246.250.214] DOI: https://doi.org/10.4103/mtsp.mtsp_4_22.

Abbreviations

PCOS:

Polycystic ovary syndrome

VDR:

Vitamin D receptor

RT-PCR:

Real-time polymerase chain reaction

IR:

Insulin resistance

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Acknowledgements

The authors are highly thankful for the support and amenities provided by the Centre of Research for Development (CORD) at the University of Kashmir, Srinagar and the Department of Endocrinology at Superspeciality Hospital Srinagar, Kashmir. A. Ashraf acknowledges support from Shri Venkateshwara University for carrying out Ph.D. research on PCOS, from subject Biochemistry under the Department of Applied Sciences.

Funding

This research didn’t receive any specific grant from any funding agencies in the public, commercial, or not-for-profit sectors.

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Authors and Affiliations

  1. Department of Biochemistry, School of Applied Sciences, Shri Venkateshwara University, Gajraula Amroha, UP, 244236, India

    Ahila Ashraf & Rajesh Singh

  2. Centre of Research for Development, University of Kashmir, Srinagar, J&K, 190006, India

    Ahila Ashraf & Bashir Ahmad Ganai

  3. Department of Endocrinology, Government Medical College, Super Speciality Hospital, Srinagar, J&K, India

    Shahnawaz Mir

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  1. Ahila Ashraf
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Contributions

Professor (Dr.) R. Singh and A. Ashraf held the frame responsibility of the present study; A. Ashraf conducted the research, collected the patient data and information, composed the paper work and executed the statistical data analysis. Professor (Dr.) Bashir Ahmad Ganai held the main responsibility of providing the work understanding, including the theory understanding, bench work, methodological contributions and all lab work data to accomplish the practical portion of the research work. Dr. Shahnawaz Mir held the responsibility over guidance and compiling of clinico-pathological data of the patients in his presence. The intellectual content of this article was evaluated and analyzed decisively by Professor R. Singh, Professor B.A. Ganai and Dr. S. Mir. The major concern for the concluding work of this article was carried on by A. Ashraf, Professor R. Singh, Professor B.A. Ganai and Dr. S. Mir. The prime concern and aim of this research was to evaluate and perceive the study of PCOS genetically, epigenetically as well and clinico-pathologically with respect to vitamin D, so as to understand this disease over higher perspectives, which can be beneficial to the society in coming future and also to lead a study finding out if there can be some more and accurate diagnostic criteria of PCOS in near future.

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Correspondence to Ahila Ashraf.

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There has been no conflict of interest from any of the authors.

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The present research work was approved by the Institutional Ethics Committee (IEC) of GMC, Srinagar under Ref. No. 142/ETH/GMC/ICM, 2019.

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All the participants had given their written informed consent in a written in-formed consent letter. All methods were carried out in accordance with relevant guidelines and regulations.

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Ashraf, A., Singh, R., Ganai, B.A. et al. Hypermethylation and down-regulation of vitamin D receptor (VDR) as contributing factors for polycystic ovary syndrome (PCOS): a case–control study from Kashmir, North India. Arch Gynecol Obstet 309, 1091–1100 (2024). https://doi.org/10.1007/s00404-023-07326-9

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