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Cargo small non-coding RNAs of extracellular vesicles isolated from uterine fluid associate with endometrial receptivity and implantation success
Fertility and Sterility ( IF 6.6 ) Pub Date : 2020-12-01 , DOI: 10.1016/j.fertnstert.2020.10.046
Tiantian Li 1 , Ellen M Greenblatt 2 , Michelle EyunJung Shin 3 , Theodore J Brown 4 , Crystal Chan 2
Affiliation  

OBJECTIVE To optimize a method of isolating extracellular vesicles (EVs) from uterine fluid and to characterize small non-coding RNAs (sncRNAs) from the EVs, with the goal of identifying novel receptivity-associated biomarkers. DESIGN Longitudinal study comparing sncRNA expression profiles from endometrial EVs. SETTING University-affiliated, hospital-based fertility clinic. PATIENT(S) Healthy volunteers with no history of infertility (Group A) and women receiving controlled ovarian stimulation (COS)-in vitro fertilization treatment (Group B). INTERVENTIONS(S) In Group A, EVs were isolated from uterine fluid obtained on luteinizing hormone (LH)+2 and LH+7 in one natural menstrual cycle. In Group B, EVs were isolated from uterine fluid obtained on human chorionic gonadotropin (hCG)+2 and hCG+7 in one COS cycle. RNAs extracted from EVs were profiled using next-generation sequencing. MAIN OUTCOME MEASURE(S) Differential EV-sncRNAs between LH+2 and LH+7 (Group A), between hCG+2 and hCG+7 (Group B), and between pregnant and nonpregnant in vitro fertilization cycles (Group B). RESULT(S) Ultracentrifugation was validated as the most efficient method to isolate EVs from uterine fluid. We identified 12 endometrial EV-sncRNAs (11 microRNAs and 1 piwi-interacting RNA) as receptivity-associated transcripts conserved in both natural and COS cycles. These sncRNAs were associated strongly with biological functions related to immune response, extracellular matrix, and cell junction. Within COS cycles, we also identified a group of EV-sncRNAs that exhibited differential expression in patients who conceived versus those who did not, with hsa-miR-362-3p most robustly overexpressed in the nonpregnant patients. CONCLUSION(S) This study is the first to profile comprehensively sncRNAs in endometrial EVs from uterine fluid and identify sncRNA biomarkers of endometrial receptivity and implantation success.

中文翻译:

从子宫液中分离的细胞外囊泡的货物小非编码 RNA 与子宫内膜容受性和植入成功相关

目的 优化从子宫液中分离细胞外囊泡 (EV) 的方法,并表征来自 EV 的小非编码 RNA (sncRNA),目的是识别新的接受性相关生物标志物。设计 比较来自子宫内膜 EV 的 sncRNA 表达谱的纵向研究。设置 大学附属医院生育诊所。患者 没有不孕史的健康志愿者(A 组)和接受控制性卵巢刺激(COS)-体外受精治疗的女性(B 组)。干预措施(S) 在 A 组中,从在一个自然月经周期中使用促黄体生成素 (LH)+2 和 LH+7 获得的子宫液中分离出 EV。在 B 组中,在一个 COS 周期中,从人绒毛膜促性腺激素 (hCG)+2 和 hCG+7 获得的子宫液中分离出 EV。从 EV 中提取的 RNA 使用下一代测序进行了分析。主要结果测量 LH+2 和 LH+7(A 组)之间、hCG+2 和 hCG+7(B 组)之间以及怀孕和非怀孕体外受精周期(B 组)之间的差异 EV-sncRNA。结果(S)超速离心被证实是从子宫液中分离 EV 的最有效方法。我们鉴定了 12 个子宫内膜 EV-sncRNA(11 个 microRNA 和 1 个 piwi 相互作用的 RNA)作为在自然和 COS 循环中都保守的接受性相关转录本。这些 sncRNA 与免疫反应、细胞外基质和细胞连接等生物学功能密切相关。在 COS 周期内,我们还确定了一组 EV-sncRNA,这些 EV-sncRNA 在受孕患者与未受孕患者中表现出差异表达,hsa-miR-362-3p 在非妊娠患者中过表达最为强烈。结论(S)本研究首次全面分析了子宫内膜 EV 中的 sncRNA,并确定了子宫内膜容受性和植入成功的 sncRNA 生物标志物。
更新日期:2020-12-01
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