The role of endocannabinoids receptors in regulating trophoblast cell function

背景 内源性大麻素是一种基于脂质的信号分子，可调节疼痛、食欲和神经内分泌功能。最近有报道称，大麻素受体1和2（CB 1和CB 2）沿着第三种亚型GPR 55可能与生殖组织中的内源性大麻素信号有关。它们可能有助于胎盘的发育和滋养层细胞（构成胎盘的干细胞）的功能。然而，具体的作用和机制，这些受体调节的主要接口，管理母亲/胎儿的相互作用知之甚少。来自我实验室的体外证据表明，刺激培养的滋养层细胞中的内源性大麻素受体（s）大大降低了它们侵入和形成融合合胞体的能力;这是形成母胎界面的基本步骤。 假设 我们的假设是，破坏滋养层细胞中的大麻素受体信号传导将导致与胎盘发育、侵袭、血管重塑和内分泌功能相关的过程的进展改变。 研究计划 目标1。在培养的人滋养层细胞中，大麻素受体信号传导影响的功能途径的阐明。将使用人滋养层细胞进行机制研究，因为没有普遍接受的啮齿动物滋养层细胞系。我们将评估在滋养层细胞系中用受体特异性激动剂和拮抗剂激活和抑制CB 1、CB 2和GPR 55的影响。将监测滋养层功能结局，如侵袭、合胞体形成和内分泌信号传导（激素产生）。我们将使用分离的人原代滋养层细胞证实我们的观察结果。 目标二。线粒体在内源性大麻素信号传导中的作用。我们将通过各种药理学和分子方法破坏这些过程来确定线粒体自由基产生和细胞器形态变化对大麻素受体信号传导的贡献，并评估对滋养层功能的影响（目的1）。 目标3：内源性大麻素受体信号传导系统性中断对滋养层功能和胎盘形成的影响。我们将评估在妊娠期间的两个时间点激活或抑制亚型特异性大麻素受体信号通路对胎盘形成进展的影响。我们将评估滋养层浸润和血管重塑的变化，这两个过程对胎盘形成至关重要。我们将这些结果与胎儿生长相关联。 HQP的培训 我的计划促进了研究生，本科生的培训，采用综合方法进行技术培训和知识翻译。我对HQP培训的承诺超出了我的实验室，因为我开发了一个泛大学的推广计划，促进了本科生和研究生在科学传播方面的培训。

BACKGROUND Endogenous cannabinoids are lipid-based signaling molecules which regulate pain, appetite, neuroendocrine function. Recently it has been reported that cannabinoid receptors 1 and 2 (CB1and CB2), along with a third subtype, GPR55, may be linked to endocannabinoid signaling in reproductive tissues. They may contribute to the development of the placenta and the function of trophoblast cells (the stem cells which comprise the placenta). However, the specific roles and the mechanisms by which these receptors regulate the primary interface that governs maternal/fetal interactions are poorly understood. In vitro evidence from my laboratory suggests that stimulation of the endocannabinoid receptor(s) in cultured trophoblast cells substantially reduce their ability to invade and form a fused syncytium; essential steps in forming the maternal-fetal interface. HYPOTHESIS Our hypothesis is that disruption of cannabinoid receptor signaling in trophoblast cells will result in altered progression of the processes associated with placental development; invasion, vascular remodeling and endocrine function. RESEARCH PROGRAM Aim 1. Elucidation of the functional pathways impacted by cannabinoid receptor signaling in cultured human trophoblasts. Mechanistic studies will be conducted using human trophoblast cells since there are no universally accepted rodent trophoblast cell lines. We will evaluate the impact of activating and inhibiting CB1, CB2 and GPR55 with receptor specific agonists and antagonists in trophoblast cell lines. Trophoblast functional outcomes such as invasion, syncytium formation and endocrine signaling (hormone production) will be monitored. We will confirm our observations using isolated human primary trophoblasts. Aim 2. The role of mitochondria in endocannabinoid signaling. We will determine the contribution of mitochondrial free radical production and changes in organelle morphology towards cannabinoid receptor signaling by disrupting these process with various pharmacological and molecular approaches and evaluate the consequences on trophoblast function (Aim 1). Aim 3. Consequences of systemic disruption of endocannabinoid receptor signaling on trophoblast function and placentation. We will evaluate the consequence of activating or inhibition of subtype-specific cannabinoid receptor signaling pathways on the progression of placentation at two time points during gestation. We will evaluate changes in trophoblast invasion and vascular remodeling; two processes which are critical to placentation. We will associate these outcomes with fetal growth. TRAINING OF HQP My program facilitates the training of graduate, undergraduate students using an integrated approach to technical training and knowledge translation. My commitment to training HQP extends beyond my laboratory as I have developed a pan-university outreach program which facilitates undergraduate and graduate student training in science communication.