Hamshaw Isabel, Straube Anne, Stark Richard, Baxter Laura, Alam Mohammad T, Wever Walter J, Yin Jun, Yue Yong, Pinton Philippe, Sen Aritro, Ferguson Gregory D, Blanks Andrew M
Clinical Science Research Laboratories, Division of Biomedical Sciences, Warwick Medical School, University of Warwick, Coventry, United Kingdom.
Centre for Mechanochemical Cell Biology, Division of Biomedical Sciences, University of Warwick, Coventry, United Kingdom.
Front Pharmacol. 2023 Dec 14;14:1285779. doi: 10.3389/fphar.2023.1285779. eCollection 2023.
Preterm birth is the leading cause of infant morbidity and mortality. There has been an interest in developing prostaglandin F (PGF) antagonists as a new treatment for preterm birth, although much of the rationale for their use is based on studies in rodents where PGF initiates labour by regressing the corpus luteum and reducing systemic progesterone concentrations. How PGF 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 PGF receptor (FP) antagonist. We verified the functionality of the cell line by stimulation with PGF, resulting in Gα-specific coupling and Ca release, which were inhibited by FP antagonism. Compared to four published FP receptor antagonists, the novel FP antagonist N582707 was the most potent compound [F 7.67 ± 0.63 (IC 21.26 nM), AUC 7.30 ± 0.32 (IC 50.43 nM), and frequency of Ca oscillations 7.66 ± 0.41 (IC 22.15 nM)]. RNA-sequencing of the MYLA cell line at 1, 3, 6, 12, 24, and 48 h post PGF treatment revealed a transforming phenotype from a fibroblastic to smooth muscle mRNA profile. PGF treatment increased the expression of , , and as well as the pro-labour genes , , and , 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 , , and . Our findings suggest that in addition to the well-described acute contractile effect, PGF 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.
早产是婴儿发病和死亡的主要原因。人们一直对开发前列腺素F（PGF）拮抗剂作为早产的新治疗方法感兴趣，尽管其使用的许多理论依据是基于在啮齿动物中的研究，在这些研究中PGF通过使黄体退化和降低全身孕酮浓度来启动分娩。PGF拮抗作用在全身孕酮不下降的人类中如何起作用仍不清楚。除了急性刺激宫缩外，一种可能性是子宫肌层平滑肌细胞状态直接改变为促分娩表型。在本研究中，我们建立了一种永生化子宫肌层细胞系MYLA，它源自一名未分娩的孕妇的子宫肌层组织，以及一类新型的PGF受体（FP）拮抗剂。我们通过用PGF刺激来验证细胞系的功能，导致Gα特异性偶联和钙释放，而这被FP拮抗作用所抑制。与四种已发表的FP受体拮抗剂相比，新型FP拮抗剂N582707是最有效的化合物[F 7.67±0.63（IC21.26 nM），AUC 7.30±0.32（IC50.43 nM），以及钙振荡频率7.66±0.41（IC22.15 nM）]。在PGF处理后1、3、6,、12、24和48小时对MYLA细胞系进行RNA测序，揭示了从成纤维细胞型到平滑肌mRNA谱的转变表型。PGF处理增加了、和的表达以及促分娩基因、和的表达，而这些被FP拮抗作用所抑制。与抑制平滑肌、促分娩转变同时，FP拮抗作用增加了成纤维细胞标记基因、和的表达。我们的研究结果表明，除了众所周知的急性收缩作用外，PGF还将子宫肌层平滑肌细胞从肌成纤维细胞转变为平滑肌、促分娩样状态，并且新型化合物N582707除了用作急性宫缩抑制剂外，还具有在早产管理中预防性使用的潜力。
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