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miR-4433a-3p promotes granulosa cell apoptosis by targeting peroxisome proliferator–activated receptor alpha and inducing immune cell infiltration in polycystic ovarian syndrome

  • Reproductive physiology and disease
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Background

Granulosa cell (GC) proliferation and apoptosis are critical events of the ovum energy supply, which lead to follicular growth retardation or atresia, and various ovulatory obstacles, eventually resulting in the development of ovarian disorders such as polycystic ovarian syndrome (PCOS). Apoptosis and dysregulated miRNA expression in GCs are manifestations of PCOS. miR-4433a-3p has been reported to be involved in apoptosis. However, there is no study reporting the roles of miR-4433a-3p in GC apoptosis and PCOS progression.

Methods

miR-4433a-3p and peroxisome proliferator–activated receptor alpha (PPAR-α) levels in GCs of PCOS patients or in tissues of a PCOS rat model were examined by quantitative polymerase chain reaction and immunohistochemistry. Bioinformatics analyses and luciferase assays were used to examine the association between miR-4433a-3p and PPAR-α, as well as PPAR-α and immune cell infiltration, in PCOS patients.

Results

miR-4433a-3p expression in GCs of PCOS patients was increased. miR-4433a-3p overexpression inhibited the growth of the human granulosa-like tumor cell line (KGN) and promoted apoptosis, while co-treatment with PPAR-α and miR-4433a-3p mimic rescued miR-4433a-3p-induced apoptosis. PPAR-α was a direct target of miR-4433a-3p and its expression was decreased in PCOS patients. PPAR-α expression was also positively correlated with the infiltration of activated CD4+ T cells, eosinophils, B cells, gamma delta T cells, macrophages, and mast cells, but negatively correlated with the infiltration of activated CD8+ T cells, CD56+ bright natural killer cells, immature dendritic cells, monocytes, plasmacytoid dendritic cells, neutrophils, and type 1 T helper cells in PCOS patients.

Conclusion

The miR-4433a-3p/PPAR-α/immune cell infiltration axis may function as a novel cascade to alter GC apoptosis in PCOS.

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Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request, and in the Gene Expression Omnibus (GEO) repository. GSE84376 (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE84376).

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Funding

This research was supported by the National Natural Science Foundation of China (No. 81801532).

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Author notes

  1. Lin Zhu, ** Yao, and Ying Mo contributed equally to this work.

Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Center for Reproductive Medicine, Guangdong Provincial Key Laboratory of Major Obstetric Diseases, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, China

    Lin Zhu, ** Yao, Ying Mo, Ming-wei Chen, Si-chen Li, Jian-qiao Liu & Hai-ying Liu

  2. Key Laboratory for Reproductive Medicine of Guangdong Province, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, China

    Lin Zhu, ** Yao, Ying Mo, Ming-wei Chen, Si-chen Li, Jian-qiao Liu & Hai-ying Liu

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Contributions

LZ and XY analyzed and interpreted the patient data regarding the PCOS. YM, MWC, and SCL performed the histological examination of the ovarian and animal experiment. HYL and JQL were major contributors in writing the manuscript. All authors read and approved the final manuscript.

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Correspondence to Jian-qiao Liu or Hai-ying Liu.

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All study protocols were approved by the Institutional Ethics Committee of the Third Affiliated Hospital Guangzhou Medical University (Ethics Review Board, 2018NO:083; Guangzhou, China).

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Zhu, L., Yao, X., Mo, Y. et al. miR-4433a-3p promotes granulosa cell apoptosis by targeting peroxisome proliferator–activated receptor alpha and inducing immune cell infiltration in polycystic ovarian syndrome. J Assist Reprod Genet 40, 1447–1459 (2023). https://doi.org/10.1007/s10815-023-02815-x

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