The role and regulation of cell survival in uterine functions.

细胞存活在子宫功能中的作用和调节。

• 批准号: RGPIN-2019-06151
• 负责人: Asselin, Eric
• 金额: $ 3.64万
• 依托单位: Université du Québec à Trois-Rivières
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=745045
• 关键词: role; regulation; cell; survival; uterine

BACKGROUND & RATIONALE: Prostate apoptosis response-4 (Par-4), a novel regulator of apoptosis as well as epithelial to mesenchymal transition (EMT), is active in a variety of cell lines and knockout mice show reduced lifespan and enhanced abnormal cell proliferation in the endometrium. We demonstrated that Par-4 is a direct target of caspase-3 during apoptosis induction and that the resulting cleaved fragment is capable of inducing programmed cell death through its translocation to nucleus. We also showed that TGFß increases EMT of endometrial cells and that it can regulate the long form of Par-4 in vitro. Interestingly, preliminary evidence gathered using cell lines in our laboratory and publically available databases suggest that 17ß-estradiol (E2) can negatively regulate Par-4. E2, an important female sex hormone, act through two known pathways, the canonical genomic pathway involving its intracellular receptor ER? which bind to estrogen-response elements (ERE) in DNA and the non-genomic pathway involving the PI-3K/Akt signaling action, an important survival and proliferation actor in the endometrium. How E2 action can be modulated through the inhibition of the pro-apoptotic factor Par-4 is unknown and it is crucial that we understand the molecular mechanisms controlling these functions. HYPOTHESES: 1) E2 reduces Par-4 expression and modifies its activity as well as its localization, abrogating its ability to induce apoptosis in endometrial cells. Therefore, inhibition of E2 activity should reinstate Par-4 expression and activity. This hypothesis will be explored in Obj1. 2) Par-4 is negatively regulated through the non-genomic E2 pathway or through PI3/Akt axis; this pathway would also influence Par-4 localization through TGFß signaling, which plays a pivotal role in cell survival and EMT/MET. This hypothesis will be explored in Obj2. The overall and long-term objective of the proposed research program is to examine and understand the regulation of expression as well as the endocrine, paracrine and autocrine activity of Par-4 in the regulation of endometrial cell fate. EXPERIMENTAL APPROACHES: Known estrogen responsive and non-responsive endometrial cells will be used for in vitro studies. Mouse pregnancy and induced decidualization will be used to study TGF-ß and Par-4 axis in the EMT process. SPECIFIC OBJECTIVES: 1) To determine the role of E2, through ERa and PI-3K/Akt pathway, in the control of Par-4 expression, localization and activity; 2) To elucidate how Par-4 differentially induce apoptosis and EMT and the key mechanisms governing these processes. INNOVATION & SIGNIFICANCE: A deeper understanding of E2 and TGFß-dependent Par-4 regulation is essential in uterine functions. Information gained from these studies will also help to identify the molecular mechanisms controlling apoptosis/survival and other types of cell fate (migration, invasion, EMT/MET) and will provide important clues in our understanding of reproductive functions.

背景和依据：前列腺细胞凋亡反应-4（Par-4）是一种新型的细胞凋亡调节因子，也是上皮细胞向间质细胞转化（EMT）的调节因子，在多种细胞系中具有活性，敲除小鼠的寿命缩短，子宫内膜异常细胞增殖增强。我们证明，Par-4是凋亡诱导过程中caspase-3的直接靶点，并且所产生的切割片段能够通过其易位到细胞核来诱导程序性细胞死亡。我们还表明，TGF β 1增加子宫内膜细胞的EMT，并且它可以在体外调节长型Par-4。有趣的是，使用我们实验室中的细胞系和药物可用数据库收集的初步证据表明，17 β-雌二醇（E2）可以负调节Par-4。雌激素E2是一种重要的雌性激素，其作用途径有两条，一条是细胞内受体ER参与的经典基因组途径;其与DNA中的雌激素反应元件（ERE）结合，并与涉及PI-3 K/Akt信号传导作用的非基因组途径结合，PI-3 K/Akt信号传导作用是子宫内膜中重要的存活和增殖因子。如何E2的行动可以通过抑制促凋亡因子Par-4调制是未知的，这是至关重要的，我们了解控制这些功能的分子机制。 假设：1）E2降低Par-4表达并改变其活性及其定位，消除其诱导子宫内膜细胞凋亡的能力。因此，抑制E2活性应恢复Par-4表达和活性。这个假设将在目标1中探讨。2)Par-4通过非基因组E2途径或通过PI 3/Akt轴负调控;该途径还将通过TGF β信号传导影响Par-4定位，其在细胞存活和EMT/MET中起关键作用。这个假设将在目标2中探讨。 拟议研究计划的总体和长期目标是检查和了解Par-4在调节子宫内膜细胞命运中的表达调节以及内分泌，旁分泌和自分泌活性。实验方法：已知的雌激素反应性和非反应性子宫内膜细胞将用于体外研究。本研究将利用小鼠妊娠和诱导蜕膜化研究TGF-β 1和Par-4轴在EMT过程中的作用。具体说明：1）研究E2通过ER α和PI-3 K/Akt通路调控Par-4的表达、定位和活性; 2）阐明Par-4如何差异诱导细胞凋亡和EMT，以及调控这些过程的关键机制。 创新与意义：深入了解E2和TGF β依赖的Par-4调节在子宫功能中至关重要。从这些研究中获得的信息也将有助于确定控制细胞凋亡/存活和其他类型的细胞命运（迁移，侵袭，EMT/MET）的分子机制，并将为我们理解生殖功能提供重要线索。