PGF2α induces a pro-labour phenotypical switch in human myometrial cells that can be inhibited with PGF2α receptor antagonists

Isabel Hamshaw, Anne Straube, Richard Stark, Laura Baxter, Mohammad Tauqeer Alam, Walter J. Wever, Jun Yin, Yong Yue, Philippe Pinton, Aritro Sen, Gregory D. Ferguson, Andrew M. Blanks

Research output: Contribution to journalArticlepeer-review

Abstract

Preterm birth is the leading cause of infant morbidity and mortality. There has been an interest in developing prostaglandin F2α (PGF2α) antagonists as a new treatment for preterm birth, although much of the rationale for their use is based on studies in rodents where PGF2α initiates labour by regressing the corpus luteum and reducing systemic progesterone concentrations. How PGF2α antagonism would act in humans who do not have a fall in systemic progesterone remains unclear. One possibility, in addition to an acute stimulation of contractions, is a direct alteration of the myometrial smooth muscle cell state towards a pro-labour phenotype. In this study, we developed an immortalised myometrial cell line, MYLA, derived from myometrial tissue obtained from a pregnant, non-labouring patient, as well as a novel class of PGF2α receptor (FP) antagonist. We verified the functionality of the cell line by stimulation with PGF2α, resulting in Gαq-specific coupling and Ca2+ release, which were inhibited by FP antagonism. Compared to four published FP receptor antagonists, the novel FP antagonist N582707 was the most potent compound [Fmax 7.67 ± 0.63 (IC50 21.26 nM), AUC 7.30 ± 0.32 (IC50 50.43 nM), and frequency of Ca2+ oscillations 7.66 ± 0.41 (IC50 22.15 nM)]. RNA-sequencing of the MYLA cell line at 1, 3, 6, 12, 24, and 48 h post PGF2α treatment revealed a transforming phenotype from a fibroblastic to smooth muscle mRNA profile. PGF2α treatment increased the expression of MYLK, CALD1, and CNN1 as well as the pro-labour genes OXTR, IL6, and IL11, which were inhibited by FP antagonism. Concomitant with the inhibition of a smooth muscle, pro-labour transition, FP antagonism increased the expression of the fibroblast marker genes DCN, FBLN1, and PDGFRA. Our findings suggest that in addition to the well-described acute contractile effect, PGF2α transforms myometrial smooth muscle cells from a myofibroblast to a smooth muscle, pro-labour–like state and that the novel compound N582707 has the potential for prophylactic use in preterm labour management beyond its use as an acute tocolytic drug.
Original languageEnglish
JournalFrontiers in Pharmacology
Volume14
Early online date14 Dec 2023
DOIs
Publication statusPublished - 14 Dec 2023

Acknowledgements

The authors are grateful to the women and couples who consented to this research and are indebted to all the staff in the Centre for Reproductive Medicine and Biomedical Research Unit, University Hospitals Coventry, and Warwickshire National Health Service Trust for facilitating myometrial sample collection. The authors thank Parmjit S. Jat (University College London) for providing stable retroviral producer cells for immortalisation. The authors are indebted to the staff at the Genomics Facility at the University of Warwick for running the next-generation sequencing and to Ayan Dirir, Paul J. Brighton, and Emma S. Lucas for technical assistance with the research.

Funding

The authors declare financial support was received for the research, authorship, and/or publication of this article. This work was funded by a project grant awarded to AB from Ferring Research Institute Inc. (San Diego, CA, United States). GF co-conceived the project. AS was supported by the Wellcome Trust Investigator Award (200870/Z/16/Z).

FundersFunder number
The Wellcome Trust

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