Targeting cellular senescence to inhibit the development and progression of ovarian endometriomas

靶向细胞衰老抑制卵巢子宫内膜异位症的发生和进展

• 批准号: 10512390
• 负责人: Shannon Michelle Hawkins
• 金额: $ 65.72万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-15 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10512390
• 关键词: ARID1A gene; Address; Adhesions; Adult; Affect; Assisted Reproductive Technology; Automobile Driving; Biological; Biological Markers; CDKN2A gene; Cell Aging; Cell Culture Techniques; Cell Cycle Arrest; Cell Differentiation process; Cells; Cellular Morphology; Clinical; Clinical Research; Data; Development; Developmental Process; Disease; Endometriomas; Enterobacteria phage P1 Cre recombinase; Excision; Exhibits; Fertility; Fertility Rates; Future; Ganciclovir; Genes; Genetic Engineering; Genetic Recombination; Goals; Gynecologic; HSV-Tk Gene; Harvest; Health; Histologic; Human; In Vitro; Individual; Inflammatory; KRAS oncogenesis; KRASG12D; Lesion; Luciferases; Mediating; Modeling; Molecular; Mus; Mutation; National Institute of Child Health and Human Development; Operative Surgical Procedures; Ovarian; Ovarian Clear Cell Tumor; Ovary; Pancreas; Pathogenesis; Pelvic cavity structure; Pharmacology; Phenotype; Pregnancy Outcome; Process; Reproducibility; Reproductive Health; Research; Research Personnel; Retrograde Menstruation; Role; Signal Transduction; Strategic Planning; Surgical complication; Testing; Time; Woman; beta-Galactosidase; cell type; cellular targeting; endometriosis; granulosa cell; high risk; hormone therapy; improved; molecular marker; mouse model; novel; reproductive; senescence; surgical risk; theories; therapeutic target; transcriptome; transcriptomics; transdifferentiation

Ovarian endometriomas are deep endometriosis lesions on the ovary. Endometriomas are a unique form of endometriosis in that they do not respond to hormonal therapy and carry the highest risk of developing clear cell ovarian cancer. There is an urgent need to determine the unique pathogenesis of endometriomas to improve the lives of women. As an alternative to retrograde menstruation, the induction theory of endometriosis posits that a substance induces an adult cell to transdifferentiate into endometriosis, although the inductive substances or the cells which transdifferentiate into endometriosis have yet to be identified. AKA mice (Arid1aflox/flox; Krasflox-stop-flox-G12D; Amhr2Cre) spontaneously and reproducibly develop large, cystic endometriomas that recapitulate human endometriomas at the histologic and molecular level. As the genetic recombination lies in the granulosa cells of the ovary, AKA endometriomas do not develop by retrograde menstruation. The AKA mouse model allows for the cellular and molecular interrogation of the paradigm- shifting induction theory of endometriosis using a rigorously reproducible and easily manipulatable model. Transcriptomic analysis of AKA endometriomas revealed enrichment in cellular senescence genes. Cellular senescence is defined as a permanent cell cycle arrest. Senescent cells exhibit a senescence- associated secretory phenotype (SASP) and secrete high levels of pro-inflammatory molecules, similar to those found in the pelvic cavity of women with endometriosis. These results suggest that senescent cells in the AKA ovary secrete factors and induce endometriosis. As for the cells induced, granulosa cells exhibit the ability to transdifferentiate, a developmental process by which fully differentiated cells change into different fully differentiated cells. The role of senescence in endometriomas is conceptually novel, and the ability of granulosa cells to transdifferentiate into endometriosis through senescence signaling is a new paradigm. The central hypothesis is that the senescent microenvironment, mediated by Arid1a loss and oncogenic Kras, is critical for developing endometriomas through induction and transdifferentiation of granulosa cells. The objective of Aim 1 is to characterize the unique transcriptomic profile of the senescent cells and the endometriotic microenvironment using spatial transcriptomics. The objectives of Aim 2 are to identify the genetic changes (i.e., Kras G12D) required for senescence in granulosa cells, validate the expression of the endometriosis SASP (from SA 1), and establish SASP-mediated endometriosis transdifferentiation using primary murine granulosa cell cultures and a soluble Cre recombinase. The objectives of Aim 3 are to determine if senescence is essential for endometriosis and determine whether senotherapies restore ovarian function, fertility, and reduce endometrioma development and progression. Targeting senescence through senotherapies is critical to developing non- hormonal therapies, an urgent unmet need for endometriosis.

卵巢子宫内膜异位瘤是卵巢深部子宫内膜异位病变。子宫内膜异位症是一种独特的形式 子宫内膜异位症，因为它们对激素治疗没有反应，并且发生明确子宫内膜异位症的风险最高 细胞卵巢癌。迫切需要确定子宫内膜异位症的独特发病机制 改善妇女的生活。作为逆行月经的替代方案，子宫内膜异位症的诱导理论 假设某种物质诱导成体细胞转分化为子宫内膜异位症，尽管诱导性 转分化为子宫内膜异位症的物质或细胞尚未确定。又名小鼠 （Arid1aflox/flox；Krasflox-stop-flox-G12D；Amhr2Cre）自发且可重复地发育成大的囊状 子宫内膜异位瘤在组织学和分子水平上重现人类子宫内膜异位瘤。由于遗传 重组发生在卵巢的颗粒细胞中，又称为子宫内膜异位瘤，不会逆行发展 经期。 AKA 小鼠模型允许对范式进行细胞和分子询问- 使用严格可重复且易于操作的转移子宫内膜异位症诱导理论 模型。 AKA 子宫内膜异位瘤的转录组分析揭示了细胞衰老的富集 基因。细胞衰老被定义为永久性的细胞周期停滞。衰老细胞表现出衰老现象—— 相关分泌表型（SASP）并分泌高水平的促炎分子，类似 那些在患有子宫内膜异位症的女性盆腔中发现的。这些结果表明，衰老细胞 AKA卵巢分泌因子并诱发子宫内膜异位症。对于诱导的细胞，颗粒细胞表现出 转分化的能力，完全分化的细胞转变为不同的完全不同的细胞的发育过程 分化的细胞。衰老在子宫内膜异位症中的作用在概念上是新颖的，并且 颗粒细胞通过衰老信号转分化为子宫内膜异位症是一种新的方法 范例。核心假设是，衰老微环境是由 Arid1a 缺失和 致癌性 Kras，通过诱导和转分化对于子宫内膜异位症的发展至关重要 颗粒细胞。目标 1 的目标是表征衰老细胞的独特转录组谱 使用空间转录组学研究细胞和子宫内膜异位微环境。目标的目标 2 是确定颗粒衰老所需的遗传变化（即 Kras G12D） 细胞，验证子宫内膜异位症 SASP（来自 SA 1）的表达，并建立 SASP 介导的 使用原代小鼠颗粒细胞培养物和可溶性 Cre 进行子宫内膜异位症转分化 重组酶。目标 3 的目标是确定衰老是否是子宫内膜异位症所必需的 并确定 sentherapy 是否可以恢复卵巢功能、生育能力并减少子宫内膜异位症 发展和进步。通过感觉疗法靶向衰老对于发展非衰老细胞至关重要 激素疗法，子宫内膜异位症的迫切未满足需求。