Ovarian Inflammaging as a Mechanism for Ovarian Cancer

项目总结 高级别浆液性卵巢癌(HGSOC)是女性生殖系统最致命的癌症，具有 不到50%的患者在确诊后5年内存活。女人一生中排卵的总数是 卵巢癌的主要危险因素，以及抑制排卵的因素，包括口服 避孕药可显著降低HGSOC的发病率。尽管与排卵有关，HGSOC通常 在绝经后不再排卵的妇女中表现为晚期转移性癌症。因此，a 该领域的主要问题是：女性生殖旺季和卵巢老化后如何排卵？ 更年期导致卵巢癌的发生。在这里，我提出了卵巢老化的两个不同方面 促进HGSOC启动和发病的微环境。首先是卵泡液，它会变成纤维- 高生育年龄的炎症性疾病，引发输卵管上皮(FTE)肿瘤。 其次，老化的卵巢细胞外基质(ECM)的组成和生物力学的变化允许 增强肿瘤细胞的黏附和侵袭能力。最近的数据表明，HGSOC肿瘤起源于FTE，并且 暴露于卵泡液中会导致体内和体外FTE细胞的DNA损伤。然而，直接的影响是 生殖衰老对FTE肿瘤发生的影响还没有得到研究。我们实验室的工作发现了一系列 随着生育年龄的增加，人卵泡液中纤维炎性细胞因子的增加，包括 血管内皮生长因子在HGSOC转移中起作用，并参与其他器官系统的肿瘤启动。在目标1中 在这项提议中，我将检验卵泡液诱导FTE在 以年龄依赖的方式，通过候选细胞因子血管内皮生长因子的作用。我会用人类来治疗FTE细胞系 卵泡液通过老化连续体或重组人血管内皮生长因子的剂量曲线来确定对 细胞增殖、DNA损伤、氧化应激和全球基因表达。在FTE中启动肿瘤后， 肿瘤细胞迁移到卵巢进行扩张。我们实验室的工作已经表征了 随着生育年龄的增加，卵巢细胞外基质的组成，包括I型和III型胶原水平增加， 这会增加组织的硬度。HGSOC细胞已被证明倾向于3D富含胶原蛋白的基质进行扩张 众所周知，更坚硬的基质可以增加癌细胞的运动能力。在目标2中，我将确定 卵巢细胞外基质老化的组织环境促进HGSOC的发病 细胞外基质成分和卵巢组织硬度对HGSOC细胞黏附和侵袭的独立作用。 为此，我将在体外评估HGSOC细胞在水凝胶上的黏附和侵袭，这些水凝胶概括了硬度 以及老化卵巢的胶原蛋白丰富。此外，我还将评估HGSOC细胞的黏附和侵袭能力 可繁殖的幼年和老年脱细胞小鼠卵巢的体外实验以及体内移植后的观察 变成具有繁殖力的年轻和年老的主人。综上所述，这些研究将提供机械性的洞察 排卵和卵巢老化有助于HGSOC，并将关键途径确定为治疗靶点。

PROJECT SUMMARY High-grade serous ovarian carcinoma (HGSOC) is the most lethal cancer of the female reproductive system with less than 50% of patients surviving 5 years after diagnosis. A woman’s total lifetime number of ovulations is a key risk factor for developing ovarian cancer, and factors that repress ovulation including use of oral contraceptives significantly reduce HGSOC incidence. Despite the association with ovulation, HGSOC typically presents as late-stage, metastatic cancer in post-menopausal women who are no longer ovulating. Therefore, a major question in the field is: how do ovulation during a woman’s reproductive prime and the aging ovary post- menopause contribute to ovarian cancer development. Here, I propose two distinct aspects of the aging ovarian microenvironment that promote HGSOC initiation and pathogenesis. First, follicular fluid, which becomes fibro- inflammatory with advanced reproductive age, triggers tumor initiation in the fallopian tube epithelium (FTE). Second, changes to the composition and biomechanics of the aging ovarian extracellular matrix (ECM) allow for enhanced tumor cell adhesion and invasion. Recent data suggest that HGSOC tumors arise from the FTE, and exposure to follicular fluid results in DNA damage in FTE cells both in vivo and in vitro. However, the direct effect of reproductive aging on FTE tumorigenesis has not been examined. Work from our lab has identified a series of fibro-inflammatory cytokines that increase in human follicular fluid with advanced reproductive age, including VEGF which has roles in HGSOC metastasis and is involved in tumor initiation in other organ systems. In Aim 1 of this proposal, I will test the hypothesis that follicular fluid induces neoplastic transformation of the FTE in an age-dependent manner, through the action of candidate cytokine VEGF. I will treat FTE cell lines with human follicular fluid across an aging continuum or a dose-curve of recombinant-human VEGF to determine effects to cell proliferation, DNA damage, oxidative stress, and global gene expression. After tumor initiation in the FTE, tumor cells migrate to the ovary for expansion. Work from our lab has characterized significant changes to the composition of the ovarian ECM with advanced reproductive age including increased levels of collagen I and III, which increase tissue stiffness. HGSOC cells have been shown to favor a 3D collagen-rich matrix for expansion and stiffer matrices are known to increase cancer cell motility. In Aim 2, I will determine the mechanism by which the tissue environment of the aging ovarian ECM promotes HGSOC pathogenesis by investigating the independent functions of ECM composition and ovarian tissue stiffness on HGSOC cell adhesion and invasion. To this end, I will assess in vitro HGSOC cell adhesion and invasion on hydrogels that recapitulate the stiffness and collagen abundance of the aging ovary. Furthermore, I will evaluate HGSOC cell adhesion and invasion in reproductively young and old decellularized mouse ovaries ex vivo, as well as an in vivo after transplantation into reproductively young and old hosts. Taken together, these studies will provide mechanistic insight into how ovulation and the aging ovary contribute to HGSOC and identify key pathways as therapeutic targets.