Human myometrial transcriptome and DNA methylome of testosterone treated patients appear more similar to myometrium from fibroid patients [array]

Uterine fibroids, or leiomyomas, are noncancerous tumors of the myometrium and the most common reproductive tract tumors in women, with an incidence of up to 80% by age 50. Currently, hysterectomy is the only definitive cure, and effective non-hormonal therapeutics are lacking. Understanding the etiology of fibroids may lead to alternative, less invasive treatments. Several obstetric disorders, including polycystic ovary syndrome (PCOS), have been linked to uterine fibroids, and women with PCOS often exhibit hormonal imbalances, particularly elevated serum testosterone levels. However, the impact of testosterone on the myometrium remains poorly understood. We hypothesize that elevated testosterone may increase the risk of developing uterine fibroids. Using RNA sequencing and MethylationEPIC array analysis, we compared myometrial tissue from women without fibroids (MyoN, n=33), with fibroids (MyoF, n=66), and after testosterone therapy (MyoT, n=7). The transcriptomic and methylation profiles of MyoT clustered with MyoF and were distinct from MyoN. We identified 1,321 differentially expressed protein-coding genes between MyoT and MyoN, while only 494 were found between MyoT and MyoF. Disease ontology analysis of MyoT vs. MyoN revealed enrichment of the fibroid tumor gene set. Fibroid associated genes including TGFβ3, CCND1, SERPINE1, and FGFR1 were upregulated in MyoT and MyoF samples compared to MyoN samples. The DNA methylation profiles of MyoT were closer to those of MyoF, but no correlation was observed between methylation status and gene expression. Our preliminary data suggest that testosterone induces transcriptional and methylation changes in the myometrium consistent with those observed in MyoF tissues. These findings suggest that elevated testosterone may be associated with an increased risk of developing uterine fibroids. Fibroid-free myometrium (MyoN, n=33), myometrium from fibroid patient (MyoF, n=63) and myometrium from patient treated with testosterone (MyoT, n=7) were sent for Infinium Methylation Array Hybridization. C: Caucasian, AA: Black/African American, H: Hispanic, A: Asian, Multi: Multi-race

子宫肌瘤（uterine fibroids）又称平滑肌瘤（leiomyomas），是起源于子宫肌层（myometrium）的良性肿瘤，也是女性最常见的生殖道肿瘤，50岁前的发病率可达80%。目前，子宫切除术（hysterectomy）是唯一的根治性治疗手段，且尚无有效的非激素治疗方案。阐明子宫肌瘤的病因学，有望开发出侵入性更低的替代疗法。多项产科疾病（包括多囊卵巢综合征（PCOS））均与子宫肌瘤存在关联，而多囊卵巢综合征患者常出现激素失衡，尤以血清睾酮（serum testosterone）水平升高为显著特征。然而，睾酮对子宫肌层的影响目前仍不甚明确。本研究提出假设：血清睾酮水平升高可能增加子宫肌瘤的发病风险。本研究采用RNA测序（RNA sequencing）与MethylationEPIC芯片阵列分析，对三组女性的子宫肌层组织进行了对比分析：无肌瘤者的子宫肌层组织（MyoN，n=33）、肌瘤患者的子宫肌层组织（MyoF，n=66）以及经睾酮治疗后的子宫肌层组织（MyoT，n=7）。结果显示，MyoT的转录组与甲基化谱与MyoF聚为一类，且与MyoN存在显著差异。本研究在MyoT与MyoN之间鉴定出1321个差异表达的蛋白编码基因，而在MyoT与MyoF之间仅发现494个。对MyoT vs MyoN进行疾病本体论分析发现，其显著富集子宫肌瘤相关基因集。与MyoN相比，MyoT与MyoF样本中子宫肌瘤相关基因（包括转化生长因子β3（TGFβ3）、细胞周期蛋白D1（CCND1）、纤溶酶原激活物抑制剂1（SERPINE1）和成纤维细胞生长因子受体1（FGFR1））均呈上调表达。MyoT的DNA甲基化谱与MyoF更为接近，但未观察到甲基化状态与基因表达之间存在相关性。本研究的初步数据表明，睾酮可诱导子宫肌层发生转录组与甲基化组改变，且该改变与MyoF组织中观察到的变化一致。上述研究结果提示，血清睾酮水平升高可能与子宫肌瘤发病风险增加相关。本研究将无肌瘤子宫肌层组织（MyoN，n=33）、肌瘤患者子宫肌层组织（MyoF，n=63）以及睾酮治疗患者子宫肌层组织（MyoT，n=7）送检，进行Infinium甲基化芯片杂交。注：C代表高加索人种，AA代表黑人/非裔美国人，H代表西班牙裔，A代表亚裔，Multi代表多种族。