Functional Analysis of Endometrial Stem/Progenitor Cells

描述(由申请人提供)：子宫疾病是影响人类女性生活质量和发病率/死亡率的一个极其常见的因素。在美国，每年有超过35,000名妇女被诊断出患有子宫内膜癌；子宫内膜异位症影响了高达15%的育龄妇女，并经常导致不孕。此外，妊娠早期子宫衬里的准备不足被认为是导致反复自然妊娠丢失的原因之一。很明显，在大多数器官的构建和维护过程中，干细胞协调发育和组织更新过程。然而，几乎没有关于干细胞在子宫发育过程中的研究，子宫可能是成年哺乳动物中经历最广泛的增殖性变化和发育重塑的器官。我们假设内源性子宫内膜干/祖细胞在子宫内的周期性变化中起着功能作用。我们的实验工作包括骨髓移植、外周血细胞输注和异种繁殖，共同排除了其他人所建议的骨髓为子宫组织结构贡献干细胞的可能性。利用H2 B：GFP转基因小鼠进行的脉冲追逐实验和命运定位研究表明，在离间质-肌层界面最近的腺上皮和间质隔室中存在长期标记保留细胞(LRC)。子宫内膜退化和再生的小鼠模型显示，尽管子宫蜕膜后腔上皮完全丧失，但残留腺组织足以完全取代所有上皮组织，这一事件发生在没有卵巢衍生类固醇激素的情况下。正在进行的分离和体内移植的研究将测试这些假定的基质和上皮性成体干细胞的干细胞活性。这项提议的长期目标是建立一个框架，在我们开始了解子宫内膜干细胞如何有助于正常的子宫发育和生育，以及当干细胞功能出现问题时，子宫内膜异位症和子宫内膜癌等子宫疾病的基础上建立一个框架。 公共卫生相关性：子宫功能障碍是影响人类女性生育力、生活质量和发病率/死亡率的共同因素。在美国，每年有超过3.5万名女性被诊断出患有子宫内膜癌(在女性中排名第四)，子宫内膜异位症是另一种使人衰弱的子宫疾病，经常导致严重的盆腔疼痛和不孕，在所有育龄妇女中，高达15%的人受到影响。据估计，所有哺乳动物受孕的25%-60%都会发生反复妊娠丢失，这被认为部分是由于怀孕早期子宫衬里准备不当所致。我们对这些子宫病理，特别是其病因的了解充其量是有限的。正常的子宫功能，从而成功怀孕，依赖于正常的子宫内膜生长、分化、退化(月经)和再生周期。我们假设干细胞负责维持这一子宫周期，并建议分离和鉴定子宫内膜干细胞。我们提议的研究的长期目标将是证明子宫干细胞功能缺陷与妇科疾病之间的因果联系，这些疾病对女性的生育能力和生活质量产生负面影响。

DESCRIPTION (provided by applicant): Uterine disease is an extremely common factor affecting the quality of life and morbidity/mortality of the human female. Each year, more than 35,000 women in the US are diagnosed with endometrial cancer; and endometriosis affects up to 15% of reproductive aged women and often results in infertility. Further, inadequate preparation of the uterine lining during early gestation is thought to contribute to recurrent spontaneous pregnancy loss. It is clear that stem cells coordinate developmental and tissue renewal processes during the construction and maintenance of most organs. Yet there are almost no studies of stem cells in the development of the uterus, an organ that undergoes perhaps the most extensive proliferative changes and developmental remodeling in adult mammals. We hypothesize that endogenous endometrial stem/progenitor cells play a functional role in the cyclic changes within the uterus. Our experimental efforts involving bone marrow transplantation, peripheral blood cell infusions and parabiosis have collectively ruled out the likelihood that bone marrow contributes stem cells to uterine tissue architecture as has been suggested by others. Pulse- chase experiments using H2B:GFP transgenic mice and fate mapping studies revealed the identity of long-term label retaining cells (LRC) in the glandular epithelium and stromal compartment nearest the stromal:myometrial interface. A mouse model of endometrial regression and regeneration reveals that despite complete loss of the luminal epithelium following uterine decidualization, residual glandular tissue is sufficient to completely replace all epithelial tissue, an event that occurs in the absence of ovarian derived steroid hormones. Ongoing studies involving isolation and in vivo transplantation will test the stem cell activity of these putative stromal and epithelial adult stem cells. The long term goal of this proposal is to establish a framework with which to build upon as we begin to understand how endometrial stem cells contribute to normal uterine development and fertility, as well as to uterine diseases like endometriosis and endometrial cancer when stem cell function goes awry. PUBLIC HEALTH RELEVANCE: Dysfunction of the uterus is a common factor affecting fertility, the quality of life, and morbidity/mortality of the human female. Each year, more than 35,000 women in the United States are diagnosed with endometrial cancer (fourth most frequent among women), and endometriosis, another debilitating uterine disease that often results in severe pelvic pain and infertility, affects up to 15% of all reproductive aged women. Recurrent pregnancy loss, occurring in an estimated 25-60% of all mammalian conceptions, is thought to occur in part because of faulty preparation of the uterine lining during early pregnancy. Our understanding of these uterine pathologies, and in particular their etiology, is at best limited. Normal uterine function, and thus successful pregnancy, is dependent on normal cycles of endometrial growth, differentiation, regression (menses) and regeneration. We hypothesize that stem cells are responsible for maintenance of this uterine cycle and have proposed to isolate and characterize endometrial stem cells. The long-term goal of our proposed studies will be to demonstrate a causal link between faulty uterine stem cell function and gynecologic pathologies that impact negatively on fertility and quality of life in women.