The Role of Glial Cells in Chronic Pelvic Pain of Endometriosis

项目主任/主要研究者（最后，第一，中间）：Nowak，Romana A. 子宫内膜异位症是一种炎症性疾病，影响10%的妇女，但目前在高达70%的 盆腔疼痛的女性和30%的不孕症女性。子宫内膜异位症的治疗方法 仅在美国就有220亿美元，不包括相关的慢性疾病。因此可以肯定地说， 子宫内膜异位症是女性最昂贵的生殖疾病之一，不仅在治疗方面， 也会影响患者的生产力和生活质量。参与的机制 持续性或慢性疼痛的发展和维持尚未完全了解。一个潜在 机制是“中枢致敏”的过程，长期持续的分子变化促进 中枢神经系统（CNS）内伤害感受神经元的神经可塑性，导致放大 疼痛信号。中枢敏感化的发展涉及神经元和神经胶质细胞的变化 人口。子宫内膜异位症的神经变化已经被研究过， 归因于子宫内膜异位症可能涉及导致中枢致敏的神经元过程。 然而，神经胶质细胞的潜在作用还有待研究。我们的假设是 子宫内膜异位症会表现出脑内胶质细胞的活化状态和功能的显著变化 和脊髓对来自活化巨噬细胞的持续促炎信号的反应， 异位病变和腹膜液，这种激活有助于慢性 内脏疼痛本探索性研究提案的目标是开始使用 子宫内膜异位症的体内小鼠模型，以确定是否存在子宫内膜异位病变和 随后激活腹膜中的促炎巨噬细胞， 损伤引起胶质细胞增殖、活化状态和基因表达的显著改变， 脊髓和大脑的细胞。我们将在两个具体目标中检验这一假设： 1）：表征脑和脊髓中神经胶质细胞的活化和增殖的变化 发生子宫内膜异位症的小鼠脊髓，并将这些变化与子宫内膜异位症的发生相关 慢性内脏疼痛这将通过全面的免疫组织化学分析来确定， 以及一组评估疼痛的行为测试。2）、 测序和鉴定脑小胶质细胞全基因组转录改变， 子宫内膜异位症小鼠脊髓。我们将利用单细胞RNAseq（scRNAseq）技术 分析脊髓中纯化的神经胶质细胞群体中基因表达随时间的变化 和大脑，并将这些变化与患有神经退行性病变的小鼠进行比较。 接受了假移植手术 PHS 398/2590（Rev.06/09）

Program Director/Principal Investigator (Last, First, Middle): Nowak, Romana A. Endometriosis is an inflammatory disease that affects 10% of women but is present in up to 70% of women with pelvic pain and 30% of women with infertility. The estimated cost of endometriosis is over $22 billion in the US alone, not counting associated chronic illnesses. Thus it is safe to say that endometriosis is one of the costliest reproductive diseases in women not only in terms of treatment but also lost productivity and quality of life in affected patients. The mechanisms involved in the development and maintenance of persistent or chronic pain are not fully understood. One potential mechanism is the process of ‘central sensitization’, whereby long lasting molecular changes promote neuroplasticity of nociceptive neurons within the central nervous system (CNS) resulting in amplification of pain signaling. The development of central sensitization involves changes to neuronal and glial cell populations. Neural changes have been studied in endometriosis and it has been suggested that pain attributed to endometriosis is likely to involve neuronal processes leading to central sensitization. However, a potential role for glia has yet to be investigated. Our hypothesis is that mice with endometriosis will show significant changes in the activation state and function of glial cells in the brain and spinal cord in response to persistent pro-inflammatory signaling from activated macrophages in ectopic lesions and peritoneal fluid and that this activation contributes to the development of chronic visceral pain. The goal of this exploratory research proposal is to begin testing this hypothesis using an in vivo mouse model of endometriosis to determine whether the presence of endometriotic lesions and the subsequent activation of pro-inflammatory macrophages in the peritoneum and endometriotic lesions causes substantial alterations in the proliferation, activation status and gene expression in glial cells of the spinal cord and brain. We will test this hypothesis in two specific aims: 1): To characterize changes in activation and proliferation of glial cells in the brain and spinal cord of mice that develop endometriosis and correlate these changes with development of chronic visceral pain. This will be determined by comprehensive immunohistochemical analyses of brains and spinal cords of mice with lesions as well as a panel of behavior tests that assess pain. 2): To sequence and identify genome-wide transcriptional alterations in microglia of the brain and spinal cord of mice with endometriosis. We will utilize single cell RNAseq (scRNAseq) technology to analyze changes in gene expression over time in purified populations of glial cells in spinal cords and brains of mice bearing endometriotic lesions and compare these changes with mice that have undergone sham transplantation surgery. PHS 398/2590 (Rev. 06/09) Page Continuation Format Page