Li Lingchuan, Ye Kefan, Wang Dongjie
Department of Gynaecology, The First People's Hospital of Yunnan Province, Kunming, 650032, Yunnan, China.
The Affiliated Hospital of Kunming University of Science and Technology, Kunming, 650032, Yunnan, China.
In Vitro Cell Dev Biol Anim. 2023 Mar;59(3):166-178. doi: 10.1007/s11626-023-00760-8. Epub 2023 Apr 5.
The present study identified a novel upstream long chain non-coding (lncRNA) NEAT1/miR-141-3p/HTRA1 axis that regulated the activation of NLR family pyrin domain containing 3 (NLRP3) inflammasome to modulate endometriosis (EM) development. Specifically, clinical data suggested that the expression of NLRP3 and apoptosis-associated speck-like protein containing CARD (ASC), the cleavage of caspase-1 and gasdermin D (GSDMD), and the production of inflammatory cytokines (interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, and IL-18) were all significantly increased in the ectopic endometrium (EE) tissues, compared to the normal endometrium (NE) tissues. Then, through analyzing the datasets from GEO database (GSE2339, GSE58178, and GSE7305) using the GEO2R bioinformatics tools, we verified that HtrA Serine Peptidase 1 (HTRA1) was especially enriched in the EE tissues compared to the NE tissues. To further confirm the biological functions of HTRA1, HTRA1 was overexpressed or downregulated in primary human endometrial stromal cells (hESCs) isolated from NE tissues or EE tissues, respectively. The results showed that upregulation of HTRA1 activated NLRP3 inflammasome-mediated pyroptotic cell death and cellular inflammation in NE-derived hESCs, whereas silencing of HTRA1 played an opposite role in EE-derived hESCs. In addition, the lncRNA NEAT1/miR-141-3p axis was screened as the upstream regulator of HTRA1. Mechanistically, lncRNA NEAT1 sponged miR-141-3p to positively regulate HTRA1 in a competing endogenous RNA (ceRNA) mechanisms-dependent manner. The recovery experiments in hESCs from NE and EE tissues confirmed that lncRNA NEAT1 overexpression promoted NLRP3 inflammasome-mediated pyroptotic cell death through regulating the miR-141-3p/HTRA1 axis. Taken together, this study firstly uncovered the underlying mechanisms by which a novel lncRNA NEAT1/miR-141-3p/HTRA1-NLRP3 pathway contributed to the development of EM, which provided novel diagnostic and therapeutic biomarkers for this disease.
本研究鉴定出一条新的上游长链非编码（lncRNA）NEAT1/miR-141-3p/HTRA1轴，该轴调节含吡啉结构域的NLR家族3（NLRP3）炎性小体的激活，从而调控子宫内膜异位症（EM）的发展。具体而言，临床数据表明，与正常子宫内膜（NE）组织相比，异位子宫内膜（EE）组织中NLRP3和含半胱天冬酶激活和招募结构域的凋亡相关斑点样蛋白（ASC）的表达、半胱天冬酶-1和gasdermin D（GSDMD）的裂解以及炎性细胞因子（白细胞介素（IL）-1β、IL-6、肿瘤坏死因子（TNF）-α和IL-18）的产生均显著增加。然后，通过使用GEO2R生物信息学工具分析来自GEO数据库（GSE2339、GSE58178和GSE7305）的数据集，我们证实与NE组织相比，HtrA丝氨酸蛋白酶1（HTRA1）在EE组织中尤其富集。为了进一步证实HTRA1的生物学功能，分别在从NE组织或EE组织分离的原代人子宫内膜基质细胞（hESC）中过表达或下调HTRA1。结果表明，HTRA1的上调激活了NE来源的hESC中NLRP3炎性小体介导的焦亡细胞死亡和细胞炎症，而HTRA1的沉默在EE来源的hESC中发挥相反作用。此外，lncRNA NEAT1/miR-141-3p轴被筛选为HTRA1的上游调节因子。机制上，lncRNA NEAT1通过竞争性内源RNA（ceRNA）机制依赖性方式海绵化miR-141-3p以正向调节HTRA1。来自NE和EE组织的hESC中的恢复实验证实，lncRNA NEAT1的过表达通过调节miR-141-3p/HTRA1轴促进NLRP3炎性小体介导的焦亡细胞死亡。综上所述，本研究首次揭示了一条新的lncRNA NEAT1/miR-141-3p/HTRA1-NLRP3途径促进EM发展的潜在机制，为该疾病提供了新的诊断和治疗生物标志物。
