Targeting the endometrial stem cell niche inendometriosis

项目总结 患有子宫内膜异位症的妇女子宫内膜组织在宫腔外异位生长，并表现为 结果，经历了使人虚弱的慢性盆腔疼痛，高不孕率，以及对身体的破坏性影响， 精神和社会福利。子宫内膜拥有一种独特的再生能力，使其能够生长， 在女人的一生中，区分和分解成百上千次。这一潜力是 位于深基底部子宫内膜以及子宫内膜其他区域的干细胞所赋予的 上皮和间质。在转化生长因子(转化生长因子)途径中，配体如转化生长因子和 骨形态发生蛋白(BMPs)是拮抗性的，在细胞分化过程中扮演相反的角色。我们的 已发表的小鼠模型显示子宫内膜转化生长因子和骨形态发生蛋白信号在整个妊娠过程中起着关键作用。 用于产后子宫内膜再生。利用子宫内膜器质培养，我们确定了 转化生长因子信号导致骨形态发生蛋白和维甲酸两条与干细胞相关的关键信号通路被激活 酸性信号通路。子宫内膜异位症患者的异位病变过度表达骨形态发生 蛋白质受体(BMPR2)以及催化合成维甲酸的ALDH1A1和ALDH1A3酶 在许多组织类型中都是干细胞标记。骨形态发生蛋白驱动表达ALDHHI的肿瘤干细胞的扩张 卵巢中的细胞。尽管BMPs和ALDH在其他器官的干细胞功能中起着关键作用， 它们在正常子宫内膜再生和子宫内膜异位症中的作用尚不清楚。在这个R01中，我们将 验证BMP信号通过控制子宫内膜的增殖和分化而影响子宫内膜干细胞的假说 ALDH1A1+和ALDH1A3+干细胞的分化具体目标是：1)定义维甲酸 生物合成控制子宫内膜干细胞的再生和分化2) 用细胞、器官和小鼠确定BMP受体激酶抑制治疗子宫内膜异位症的疗效 子宫内膜异位症的动物模型。我们的方法将定义ALDH1A1+和ALDH1A3+细胞如何参与 通过产生和鉴定两个新的Aldh1a1creERT2-td番茄来研究子宫内膜的再生潜能 和Aldh1a3creERT2-tdTomato报告小鼠品系的体内谱系追踪和体外研究 子宫内膜器质的形态和全基因组转录研究。来自鼠标模型的数据将 用于对具有高ALDH活性的纯化的人子宫内膜进行的研究。晋级 对于子宫内膜异位症妇女的治疗选择，我们将确定BMP 2型受体BMPR2， 子宫内膜异位症。我们还将与药物发现中心合作，测试新的 在子宫内膜异位症细胞上开发BMPR2激酶抑制剂，3D有机化合物，并在小鼠模型中 子宫内膜异位症。因此，通过确定ALDH1A1和ALDH1A3作为干细胞标记物的作用 子宫内膜，并通过推进女性这种疾病的治疗选择，我们的研究将解决 NICHD妇科健康和疾病科的战略目标。

PROJECT SUMMARY Women with endometriosis develop ectopic growth of endometrial tissue outside of the uterine cavity and as a result, experience debilitating chronic pelvic pain, high rates of infertility, and damaging effects to their physical, mental, and social well-being. The endometrium holds a unique regenerative power allowing it to grow, differentiate, and break down hundreds of times over the course of a woman’s lifetime. This potential is conferred by stem cells residing in the deep basalis endometrium, as well as in other areas of the endometrial epithelium and stroma. In the transforming growth factor  (TGF) pathway, ligands such as TGF and the bone morphogenetic proteins (BMPs) are antagonistic and play opposing roles in cell differentiation. Our published mouse models show that endometrial TGF and BMP signaling are critical throughout pregnancy and for post-partum endometrial regeneration. Using endometrial organoid cultures, we identified that loss of TGF signaling resulted in activation of two key stem-cell related signaling pathways, the BMP and retinoic acid signaling pathways. Ectopic lesions in women with endometriosis overexpress the bone morphogenetic protein receptor (BMPR2) and the ALDH1A1 and ALDH1A3 enzymes, which catalyze retinoic acid synthesis and are stem cell markers in many tissue types. BMPs drive the expansion of ALDHHI-expressing cancer stem cells in the ovary. Despite the critical roles of BMPs and ALDH on stem cell function in other organs, their roles in normal endometrial regeneration and endometriosis remain unknown. In this R01, we will examine the hypothesis that BMP signaling affects endometrial stemness by controlling the proliferation and differentiation of ALDH1A1+ and ALDH1A3+ stem cells. The Specific Aims are, 1) Define how retinoid biosynthesis controls endometrial stem cell regeneration and differentiation in the endometrium and 2) Determine the efficacy of BMP receptor kinase inhibition in endometriosis using cells, organoids, and mouse models of endometriosis. Our approach will define how ALDH1A1+ and ALDH1A3+ cells contribute to the regenerative potential of the endometrium by generating and characterizing two new Aldh1a1creERT2-tdTomato and Aldh1a3creERT2-tdTomato reporter mouse lines using in vivo lineage tracing studies and in vitro morphological and genome wide transcriptomic studies in endometrial organoids. Data from mouse models will be leveraged with studies performed in purified human endometrium with high ALDH activity. To advance therapeutic options for women with endometriosis, we will define the role of the BMP type 2 receptor, BMPR2, in endometriosis. We will also collaborate with the Center for Drug Discovery to test the efficacy of newly developed BMPR2 kinase inhibitors on endometriosis cells, 3D organoids, and in a mouse model of endometriosis. Thus, by defining the roles of ALDH1A1 and ALDH1A3 as stem cell markers of the endometrium and by advancing the therapeutic options for women with this disease, our studies will address the strategic goals of the Gynecological Health and Disease Branch at the NICHD.