CircDDX21 alleviates trophoblast dysfunction and Treg differentiation in recurrent spontaneous abortion via miR-520a-5p/ FOXP3/PD-L1 axis.

IF 3.2 3区医学 Q2 GENETICS & HEREDITY Journal of Assisted Reproduction and Genetics Pub Date : 2024-10-14 DOI:10.1007/s10815-024-03281-9

Biao Duan, Qing Feng, Li Li, Jiangfang Huang

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Abstract

Background: Recurrent spontaneous abortion (RSA) is a common complication during pregnancy, which is a burden to patients both physically and mentally. Circular RNAs (circRNAs) play important roles in RSA. However, the roles of circDDX21 in RSA development remain unknown.

Methods: Decidual samples were harvested from healthy pregnant women and RSA patients. In HTR-8/SVneo and Bewo trophoblast cells, proliferation and migration were analyzed by cell counting kit-8 (CCK-8)/5-ethynyl-2'-deoxyuridine (EdU) staining and transwell/wound healing assays, respectively. CD4⁺ T cells from peripheral blood mononuclear cells of patients were incubated with trophoblast-conditioned medium. Regulatory T cells (Treg) proliferation was detected by carboxyfluorescein succinimidyl ester (CFSE) assay. Treg proportion, Treg/T helper 17 cells (Th17) ratio, and cytokines were measured using flow cytometry. The association among genes was validated using dual-luciferase assay, RNA immunoprecipitation (RIP), and chromatin immunoprecipitation (ChIP).

Results: CircDDX21 and Forkhead box P3 (FOXP3) decreased, while miR-520a-5p increased in the decidual tissues of RSA patients. CircDDX21 overexpression promoted trophoblast proliferation and migration, and facilitated CD4⁺ T cell differentiation into Treg. CircDDX21 targeted miR-520a-5p to elevate FOXP3. MiR-520a-5p overexpression reversed the promoted trophoblast cell function of circDDX21 overexpression in HTR-8/SVneo cells. FOXP3 overexpression reversed the repressed trophoblast cell function elicited by miR-520a-5p overexpression in HTR-8/SVneo cells. FOXP3 promoted Treg differentiation by transcriptionally upregulating programmed cell death ligand 1 (PD-L1).

Conclusion: CircDDX21 ameliorated trophoblast dysfunction and Treg differentiation in RSA via miR-520a-5p/FOXP3/PD-L1 axis.

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CircDDX21通过miR-520a-5p/ FOXP3/PD-L1轴缓解复发性自然流产中滋养细胞功能障碍和Treg分化。

背景：复发性自然流产（RSA）是妊娠期常见的并发症，给患者造成身心负担。环状 RNA（circRNA）在 RSA 中发挥着重要作用。然而，circDDX21在RSA发生过程中的作用仍然未知：方法：从健康孕妇和 RSA 患者身上采集蜕膜样本。HTR-8/SVneo和Bewo滋养层细胞的增殖和迁移分别通过细胞计数试剂盒-8（CCK-8）/5-乙炔基-2'-脱氧尿苷（EdU）染色和透孔/伤口愈合试验进行分析。患者外周血单核细胞中的 CD4+ T 细胞与滋养细胞条件培养基一起培养。通过羧基荧光素琥珀酰亚胺酯（CFSE）检测调节性 T 细胞（Treg）的增殖。流式细胞术测量了Treg比例、Treg/T辅助17细胞（Th17）比例和细胞因子。使用双荧光素酶测定、RNA免疫沉淀（RIP）和染色质免疫沉淀（ChIP）验证了基因之间的关联：结果：在RSA患者的蜕膜组织中，CircDDX21和叉头框P3（FOXP3）减少，而miR-520a-5p增加。CircDDX21的过表达可促进滋养细胞的增殖和迁移，并促进CD4+ T细胞分化为Treg。CircDDX21靶向miR-520a-5p，从而提高FOXP3。MiR-520a-5p 的过表达逆转了 HTR-8/SVneo 细胞中 circDDX21 过表达对滋养层细胞功能的促进作用。FOXP3的过表达逆转了HTR-8/SVneo细胞中miR-520a-5p过表达引起的滋养层细胞功能抑制。FOXP3通过转录上调程序性细胞死亡配体1（PD-L1）促进Treg分化：结论：CircDDX21通过miR-520a-5p/FOXP3/PD-L1轴改善了RSA的滋养细胞功能障碍和Treg分化。

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来源期刊

Journal of Assisted Reproduction and Genetics 医学-妇产科学

CiteScore

5.70

自引率

9.70%

发文量

286

审稿时长

1 months

期刊介绍： The Journal of Assisted Reproduction and Genetics publishes cellular, molecular, genetic, and epigenetic discoveries advancing our understanding of the biology and underlying mechanisms from gametogenesis to offspring health. Special emphasis is placed on the practice and evolution of assisted reproduction technologies (ARTs) with reference to the diagnosis and management of diseases affecting fertility. Our goal is to educate our readership in the translation of basic and clinical discoveries made from human or relevant animal models to the safe and efficacious practice of human ARTs. The scientific rigor and ethical standards embraced by the JARG editorial team ensures a broad international base of expertise guiding the marriage of contemporary clinical research paradigms with basic science discovery. JARG publishes original papers, minireviews, case reports, and opinion pieces often combined into special topic issues that will educate clinicians and scientists with interests in the mechanisms of human development that bear on the treatment of infertility and emerging innovations in human ARTs. The guiding principles of male and female reproductive health impacting pre- and post-conceptional viability and developmental potential are emphasized within the purview of human reproductive health in current and future generations of our species. The journal is published in cooperation with the American Society for Reproductive Medicine, an organization of more than 8,000 physicians, researchers, nurses, technicians and other professionals dedicated to advancing knowledge and expertise in reproductive biology.