TET2 as a novel epigenetic regulator for uterine function and fertility

项目总结 不孕不育和不孕不育是妇女普遍存在的问题，子宫内膜对 植入胚胎被认为是一个重要的、但知之甚少的因素。10-11 易位蛋白(Tet)是铁(II)/a-酮戊二酸(Fe(II)/a-KG)依赖的甲基胞嘧啶 双加氧酶(TET1、TET2和TET3)，通过5-甲基-4-甲基-5-甲氧基苯乙酸甲酯的反复氧化来实现活性DNA去甲基化 甲基胞嘧啶(5mC)至5-羟甲基胞嘧啶(5hmC)、5-甲酰胞嘧啶(5fC)和5-羧基胞嘧啶 (5caC)。TETS还可以影响染色质修饰和基因表达，而不依赖于酶的活性。 与其他蛋白质的相互作用，包括转录因子(如WT1)和表观遗传修饰物(如SIN3A)。 新的证据表明，Tet蛋白可能是卵巢反应的关键表观遗传调节因子 类固醇激素雌激素(E_2)和孕酮(P_4)通过其同源核受体ESR1和PGR表达 对正常子宫功能的调节。我们在人类和老鼠身上的初步数据表明，这三个TET 在子宫内膜上皮细胞和间质细胞中均有表达(TET2；GT；&GT；TET3&GT；TET1)，在 子宫内膜异位症在位内膜接受窗减少。鉴于TET2在 女性肿瘤，特别是子宫内膜癌和子宫癌中顶端突变基因的研究 我们建立了子宫TET2缺失的雌性小鼠，PgrCre/+Tet2f/f(Tet2d/d)。Tet2d/d 母亲不孕不育，卵巢和输卵管功能正常。因此，这项建议的目标是 用条件性TET2突变小鼠模型确定TET2在子宫生理中的活体作用。在目标1中， 我们将确定TET2在调节感受窗中的作用。在目标2中，我们将确定 TET2在在体子宫内膜间质细胞蜕膜形成中的作用这些目标的完成将定义 TET2在子宫内膜生理调控中的表型作用，包括着床和蜕膜形成 在活体内。此外，这项鼠标工作产生的数据将为未来的标准NIH提供强有力的支持 R01/R21应用，解决子宫上皮和/或间质TET2的表观遗传机制 在怀孕期间调节子宫功能和疾病状态(例如，子宫内膜异位症和子宫内膜癌)。 因此，这项研究的发现将对人类临床医学具有重要意义。

PROJECT SUMMARY Infertility and subfertility are pervasive problems in women, and failure of the endometrial adaptation to an implanting embryo is considered a significant, yet poorly understood, contributing factor. Ten-Eleven Translocation proteins (TETs) are the iron(II)/a-ketoglutarate (Fe(II)/a-KG)-dependent methylcytosine dioxygenases (TET1, TET2 and TET3) that confer active DNA demethylation by the iterative oxidation of 5- methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC). TETs can also affect chromatin modifications and gene expression independent of enzymatic activity via interactions with other proteins including transcription factors (e.g., WT1) and epigenetic modifiers (e.g., SIN3A). Emerging evidence suggests that TET proteins may serve as key epigenetic regulators in response to ovarian steroid hormones estrogen (E2) and progesterone (P4) via their cognate nuclear receptors ESR1 and PGR in the regulation of normal uterine function. Our preliminary data in humans and mice has shown that all three TETs are expressed (Tet2>>Tet3>Tet1) in both endometrial epithelial and stromal cells, increased significantly during the window of receptivity but decreased in eutopic endometrium of endometriosis. Given that TET2 ranks among top-mutated genes in female cancers, particularly uterine corpus endometrial carcinoma (UCEC) and uterine carcinosarcoma (UCS), we have generated uterine Tet2-deleted female mice, PgrCre/+Tet2f/f (Tet2d/d). The Tet2d/d mothers were subfertile with normal ovarian and oviductal functions. Therefore, the goal of this proposal is to determine the in vivo role of Tet2 in uterine physiology using conditional Tet2 mutant mouse models. In Aim 1, we will determine the role of Tet2 in regulation of the window of receptivity. In Aim 2, we will determine the role of Tet2 in endometrial stromal cell decidualization in vivo. Completion of these aims will have defined the phenotypic roles of TET2 in the regulation of endometrial physiology, including implantation and decidualization in vivo. In addition, data generated from this mouse work will provide strong support for a future Standard NIH R01/R21 application, addressing the epigenetic mechanisms by which uterine epithelial and/or stromal TET2 regulate uterine function during pregnancy and disease states (e.g., endometriosis and endometrial cancer). Thus, the findings from this research will be important in informing human clinical medicine.