Granulosa cell-derived miR-379-5p regulates macrophage polarization in polycystic ovarian syndrome

Reza Salehi; Meshach Asare-Werehene; Brandon A Wyse; Atefeh Abedini; Bo Pan; Alex Gutsol; Sahar Jahangiri; Peter Szaraz; Kevin D Burns; Barbara Vanderhyden; Julang Li; Dylan Burger; Clifford L Librach; Benjamin K Tsang

doi:10.3389/fimmu.2023.1104550

Granulosa cell-derived miR-379-5p regulates macrophage polarization in polycystic ovarian syndrome

Front Immunol. 2023 Mar 24:14:1104550. doi: 10.3389/fimmu.2023.1104550. eCollection 2023.

Authors

Reza Salehi^{1

2

3

4}, Meshach Asare-Werehene^{1

2

3}, Brandon A Wyse⁴, Atefeh Abedini⁵, Bo Pan⁶, Alex Gutsol⁷, Sahar Jahangiri⁴, Peter Szaraz⁴, Kevin D Burns⁷, Barbara Vanderhyden^{2

3

5}, Julang Li⁶, Dylan Burger^{1

3

7}, Clifford L Librach^{4

8

9

10}, Benjamin K Tsang^{1

2

3}

Affiliations

¹ Chronic Disease Program, Ottawa Hospital Research Institute, Ottawa, ON, Canada.
² Department of Obstetrics and Gynecology, University of Ottawa, Ottawa, ON, Canada.
³ Department of Cellular and Molecular Medicine and Center for Infection, Immunity and Inflammation, University of Ottawa, Ottawa, ON, Canada.
⁴ CReATe Fertility Centre, Toronto, ON, Canada.
⁵ Cancer Therapeutics Program, Ottawa Hospital Research Institute, Ottawa, ON, Canada.
⁶ Department of Animal BioScience, University of Guelph, Guelph, ON, Canada.
⁷ Division of Nephrology, Department of Medicine, Kidney Research Centre, University of Ottawa, Ottawa, ON, Canada.
⁸ Department of Obstetrics & Gynaecology, University of Toronto, Toronto, ON, Canada.
⁹ Department of Physiology, University of Toronto, Toronto, ON, Canada.
¹⁰ Institute of Medical Sciences, University of Toronto, Toronto, ON, Canada.

Abstract

Polycystic ovarian syndrome (PCOS) is associated with hyperandrogenemia and ovarian antral follicle growth arrest. We have previously demonstrated that androgen-induced exosomal release of miR-379-5p (miR379) from preantral follicle granulosa cells increases the proliferation of target cells via phosphoinositide-dependent kinase 1 (PDK1) upregulation. Androgen also increases inflammatory M1 macrophage abundance, but reduces anti-inflammatory M2 polarization in rat antral and preovulatory follicles. However, the role of small extracellular vesicles (sEVs; also known as exosomes) secretion in determining the cellular content and function of miRNAs in exosome-receiving cells is largely unknown. Our objectives were to determine: 1) the regulatory role of granulosa cells (GC)-derived exosomal miR379 on macrophage polarization and ovarian inflammation; 2) whether miR379-induced M1 polarization regulates GC proliferation; and 3) if this regulated process is follicular stage-specific. Compared with non-PCOS subjects, PCOS subjects had a higher M1/M2 ratio, supporting the concept that PCOS is an inflammatory condition. Ovarian overexpression of miR379 increased the number of M1 macrophages and the M1/M2 ratio in preantral follicles specifically. Transfection of macrophages with a miR379 mimic reduced the cellular content of PDK1 and induced M0→M1 polarization; whereas its inhibitor polarized M0→M2. Conditioned media from macrophages transfected with miR379 mimic and follicular fluid from PCOS subjects had higher galectin-3 content, a pro-inflammatory cytokine which specifically suppresses human antral follicle GC proliferation. These results indicate that miR379 inhibits M2 macrophage polarization, a condition which suppresses GC proliferation in a follicle stage-dependent manner, as exhibited in PCOS.

Keywords: PCOS (polycystic ovarian syndrome); PDK1; exosomes; extracellular vesicle; granulosa cells; macrophage; miR-379-5p.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Androgens
Animals
Female
Granulosa Cells
Humans
Macrophages
MicroRNAs* / genetics
Polycystic Ovary Syndrome* / genetics
Rats

Substances

Androgens
MicroRNAs
MIRN379 microRNA, human

Grants and funding

MOP-119381/CIHR/Canada