The Adequate Corpus Luteum: miR-96 Promotes Luteal Cell Survival and Progesterone Production.

Bushra T Mohammed,Sadanand D Sontakke,F Xavier Donadeu,Jason Ioannidis,W Colin Duncan

doi:10.1210/jc.2017-00259

Abstract

Context:Inadequate progesterone production from the corpus luteum is associated with pregnancy loss. Data available in model species suggest important roles of microRNAs (miRNAs) in luteal development and maintenance.Objective:To comprehensively investigate the involvement of miRNAs during the ovarian follicle-luteal transition.Design:The effects of specific miRNAs on survival and steroid production by human luteinized granulosa cells (hLGCs) were tested using specific miRNA inhibitors. Candidate miRNAs were identified through microarray analyses of follicular and luteal tissues in a bovine model.Setting:An academic institution in the United Kingdom associated with a teaching hospital. hLGCs were obtained by standard transvaginal follicular-fluid aspiration from 35 women undergoing assisted conception.Intervention(s):Inhibition of candidate miRNAs in vitro.Main outcome measure(s):Levels of miRNAs, mRNAs, FOXO1 protein, apoptosis, and steroids were measured in tissues and/or cultured cells.Results:Two specific miRNA clusters, miR-183-96-182 and miR-212-132, were dramatically increased in luteal relative to follicular tissues. miR-96 and miR-132 were the most upregulated miRNAs within each cluster. Database analyses identified FOXO1 as a putative target of both these miRNAs. In cultured hLGCs, inhibition of miR-96 increased apoptosis and FOXO1 protein levels, and decreased progesterone production. These effects were prevented by small interfering RNA-mediated downregulation of FOXO1. In bovine luteal cells, miR-96 inhibition also led to increases in apoptosis and FOXO1 protein levels.Conclusions:miR-96 targets FOXO1 to regulate luteal development through effects on cell survival and steroid production. The miR-183-96-182 cluster could provide a novel target for the manipulation of luteal function.

Highlights

In monovular species, such as humans, ovulation involves rupture of the wall of a mature follicle and release of the contained oocyte for fertilization
Our studies identified a miRNA cluster which is highly expressed in the corpus luteum (CL) and plays a role in promoting luteal cell survival and steroidogenesis, providing a potential target for future interventions in human reproductive health
The miR-183-96-182 and miR-212-132 clusters are highly upregulated during the follicularluteal transition

Summary

Introduction

In monovular species, such as humans, ovulation involves rupture of the wall of a mature follicle and release of the contained oocyte for fertilization. Critical is the differentiation of estrogen-producing follicular cells into luteal cells with the ability to produce high levels of progesterone. The importance of this is reflected in the association of luteal insufficiency and/or suboptimal progesterone levels with pregnancy failure in species including cattle, sheep and horses [2,3,4]. A much-needed understanding of molecular regulation of luteal development in humans would provide clarification and assist in identifying novel therapeutic targets for fertility manipulation

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Results

Conclusion