Table_1_Obesity and PCOS radically alters the snRNA composition of follicular fluid extracellular vesicles.xlsx

IntroductionThe ovarian follicle consists of the oocyte, somatic cells, and follicular fluid (FF). Proper signalling between these compartments is required for optimal folliculogenesis. The association between polycystic ovarian syndrome (PCOS) and extracellular vesicular small non-coding RNAs (snRNAs) signatures in follicular fluid (FF) and how this relates to adiposity is unknown. The purpose of this study was to determine whether FF extracellular vesicle (FFEV)-derived snRNAs are differentially expressed (DE) between PCOS and non-PCOS subjects; and if these differences are vesicle-specific and/or adiposity-dependent.MethodsFF and granulosa cells (GC) were collected from 35 patients matched by demographic and stimulation parameters. FFEVs were isolated and snRNA libraries were constructed, sequenced, and analyzed.ResultsmiRNAs were the most abundant biotype present, with specific enrichment in exosomes (EX), whereas in GCs long non-coding RNAs were the most abundant biotype. In obese PCOS vs. lean PCOS, pathway analysis revealed target genes involved in cell survival and apoptosis, leukocyte differentiation and migration, JAK/STAT, and MAPK signalling. In obese PCOS FFEVs were selectively enriched (FFEVs vs. GCs) for miRNAs targeting p53 signalling, cell survival and apoptosis, FOXO, Hippo, TNF, and MAPK signalling.DiscussionWe provide comprehensive profiling of snRNAs in FFEVs and GCs of PCOS and non-PCOS patients, highlighting the effect of adiposity on these findings. We hypothesize that the selective packaging and release of miRNAs specifically targeting anti-apoptotic genes into the FF may be an attempt by the follicle to reduce the apoptotic pressure of the GCs and stave off premature apoptosis of the follicle observed in PCOS.

引言卵巢卵泡由卵母细胞、体细胞和卵泡液（FF）组成。这些腔室之间适当的信号传导对于卵泡发生的最佳状态至关重要。关于多囊卵巢综合征（PCOS）与卵泡液（FF）中细胞外囊泡小非编码RNA（snRNAs）特征之间的关联及其与肥胖关系尚不清楚。本研究旨在确定PCOS与非PCOS受试者之间卵泡液细胞外囊泡（FFEV）来源的snRNAs是否存在差异表达（DE）；以及这些差异是否具有囊泡特异性及/或与肥胖相关。方法从35名按人口统计学和刺激参数匹配的患者中收集FF和颗粒细胞（GC）。分离FFEV并构建snRNA文库，进行测序和分析。结果miRNAs是最丰富的生物型，在微囊泡（EX）中具有特异性富集，而在GCs中，长非编码RNA是最丰富的生物型。在肥胖PCOS与瘦PCOS的比较中，通路分析揭示了参与细胞存活和凋亡、白细胞分化和迁移、JAK/STAT和MAPK信号传导的靶基因。在肥胖PCOS中，FFEV对靶向p53信号传导、细胞存活和凋亡、FOXO、Hippo、TNF和MAPK信号传导的miRNAs进行了选择性富集（FFEVs vs. GCs）。讨论我们提供了PCOS和非PCOS患者FFEV和GCs中snRNA的全面分析，突出了肥胖对这些发现的影响。我们假设，miRNAs选择性包装和释放到FF中，专门针对抗凋亡基因，可能是卵泡为了减轻GCs的凋亡压力和防止PCOS中观察到的卵泡过早凋亡而采取的一种尝试。