Brady Paula, Yousif Abdelrahman, Sasamoto Naoko, Vitonis Allison F, Fendler Wojciech, Stawiski Konrad, Hornstein Mark D, Terry Kathryn L, Elias Kevin M, Missmer Stacey A, Shafrir Amy L
Columbia University Fertility Center, Department of Obstetrics and Gynecology, Columbia University Irving Medical Center, New York, NY, United States.
Department of Obstetrics and Gynecology, Texas Tech University Health Sciences, El Paso, TX, United States.
Front Reprod Health. 2024 Apr 11;6:1360417. doi: 10.3389/frph.2024.1360417. eCollection 2024.
Prior studies have investigated the diagnostic potential of microRNA (miRNA) expression profiles for endometriosis. However, the vast majority of previous studies have only included adult women. Therefore, we sought to investigate differential expression of miRNAs among adolescents and young adults with endometriosis.
The Women's Health Study: from Adolescence to Adulthood (A2A) is an ongoing WERF EPHect compliant longitudinal cohort. Our analysis included 64 patients with surgically-confirmed endometriosis (96% rASRM stage I/II) and 118 females never diagnosed with endometriosis frequency matched on age (median = 21 years) and hormone use at blood draw. MicroRNA measurement was separated into discovery (10 cases and 10 controls) and internal replication (54 cases and 108 controls) phases. The levels of 754 plasma miRNAs were assayed in the discovery phase using PCR with rigorous internal control measures, with the relative expression of miRNA among cases vs. controls calculated using the 2 method. miRNAs that were significant in univariate analyses stratified by hormone use were included in the internal replication phase. The internal replication phase was split 2:1 into a training and testing set and utilized FirePlex miRNA assay to assess 63 miRNAs in neural network analyses. The testing set of the validation phase was utilized to calculate the area under the curve (AUC) of the best fit models from the training set including hormone use as a covariate.
In the discovery phase, 49 miRNAs were differentially expressed between endometriosis cases and controls. The associations of the 49 miRNAs differed by hormone use at the time of blood draw. Neural network analysis in the testing set of the internal replication phase determined a final model comprising 5 miRNAs (miR-542-3p, let-7b-3p, miR-548i, miR-769-5p, miR-30c-1-3p), yielding AUC = 0.77 (95% CI: 0.67-0.87, < 0.001). Sensitivity in the testing dataset improved (83.3% vs. 72.2%) while the specificity decreased (58.3% vs. 72.2%) compared to the training set.
The results suggest that miR-542-3p, let-7b-3p, miR-548i, miR-769-5p, miR-30c-1-3p may be dysregulated among adolescent and young adults with endometriosis. Hormone use was a significant modifier of miRNA dysregulation and should be considered rigorously in miRNA diagnostic studies.
先前的研究已经探讨了微小RNA（miRNA）表达谱对于子宫内膜异位症的诊断潜力。然而，绝大多数先前的研究仅纳入了成年女性。因此，我们试图研究青少年和年轻成年子宫内膜异位症患者中miRNA的差异表达。
“从青春期到成年期女性健康研究（A2A）”是一项正在进行的符合WERF EPHect标准的纵向队列研究。我们的分析纳入了64例经手术确诊为子宫内膜异位症的患者（96%为rASRM I/II期）和118例从未被诊断为子宫内膜异位症的女性，这些女性在年龄（中位数 = 21岁）和采血时的激素使用情况方面进行了频率匹配。miRNA检测分为发现阶段（10例病例和10例对照）和内部验证阶段（54例病例和108例对照）。在发现阶段，使用带有严格内部对照措施的PCR检测了754种血浆miRNA的水平，使用2−ΔΔCT方法计算病例与对照之间miRNA的相对表达。在按激素使用情况分层的单变量分析中有显著意义的miRNA被纳入内部验证阶段。内部验证阶段按2:1分为训练集和测试集，并利用FirePlex miRNA检测法在神经网络分析中评估63种miRNA。验证阶段的测试集用于计算训练集中包括激素使用作为协变量的最佳拟合模型的曲线下面积（AUC）。
在发现阶段，49种miRNA在子宫内膜异位症病例和对照之间存在差异表达。这49种miRNA的关联因采血时的激素使用情况而异。内部验证阶段测试集中的神经网络分析确定了一个由5种miRNA（miR - 542 - 3p、let - 7b - 3p、miR - 548i、miR - 769 - 5p、miR - 30c - 1 - 3p）组成的最终模型，AUC = 0.77（95%CI：0.67 - 0.87，P < 0.001）。与训练集相比，测试数据集中的敏感性有所提高（83.3%对72.2%），而特异性有所降低（58.3%对72.2%）。
结果表明，miR - 542 - 3p、let - 7b - 3p、miR - 548i、miR - 769 - 5p、miR - 30c - 1 - 3p在青少年和年轻成年子宫内膜异位症患者中可能存在失调。激素使用是miRNA失调的一个重要调节因素，在miRNA诊断研究中应予以严格考虑。
