Identification of a Novel Target for the Treatment of Endometriosis-associated Pain

确定治疗子宫内膜异位症相关疼痛的新靶点

基本信息

批准号：
10508963
负责人：
MICHAEL SEAN ROGERS
金额：
$ 26.55万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-15 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10508963
关键词：
ARNT gene Abdominal Pain Affect Affinity Agonist American Arrestins Biological Assay CD59 Antigen Cells Chronic Disease Clinical Research Data Development Disease Drug Kinetics Drug Targeting Endometrium Estrogen Receptors Exhibits FDA approved Family Female of child bearing age Fiber Friends Functional disorder G-Protein-Coupled Receptors GTP-Binding Protein alpha Subunits, Gs GTP-Binding Proteins Gene Expression Gene Expression Profile Genes Growth Health Care Costs Hemosiderin Hormonal Human Human Genome Immune Infertility Inflammatory Knockout Mice Knowledge Laboratories Lead Lesion Libraries Lipids Measures Mediating Mediator of activation protein Methodology Modeling Molecular Non-Steroidal Anti-Inflammatory Agents Operative Surgical Procedures Pain Patients Pelvic Pain Pharmaceutical Preparations Program Development Property Publishing Receptor Cell Regimen Resources Sampling Signal Transduction Stimulus Structure Sum Surveys System Testing Therapeutic Tissues Up-Regulation Validation Woman Work antagonist cell type child bearing chronic pelvic pain cost cost estimate endometriosis experience experimental study follow-up high throughput screening human disease in vivo knock-down lipid mediator mouse model new therapeutic target novel novel therapeutics overexpression pain relief pharmacodynamic biomarker pharmacophore preclinical study receptor receptor expression screening side effect single-cell RNA sequencing success therapeutic development tool unpublished works

项目摘要

Endometriosis is an inflammatory disease characterized by the presence of endometrium-like lesions and progressive abdominal or pelvic pain. It affects ~10% of women of childbearing age and ~30% of patients are not effectively treated by existing options; new therapeutics are desperately needed. We have developed and validated a mouse model of endometriosis pain. Using that model, we discovered that the specific pro- resolving lipid mediator (SPM) protectin DX (PDX) rapidly abolishes endometriosis-associated pain and shrinks lesions. Unfortunately, poor stability and pharmacokinetic properties render PDX a poor drug lead. To leverage the anti-endometriosis activity of PDX, its receptor must be identified. All SPMs for which a high affinity receptor is known activate G-protein coupled receptors (GPCRs). Thus we hypothesize that PDX acts through a GPCR and aim to identify it and the cell type(s) expressing the PDX receptor. We will identify the mechanism by which protectin DX acts by using the PRESTO-Tango system to measure arrestin activation in a comprehensive library of human non-olfactory GPCRs. We will then identify the G-protein(s) that couple to this GPCR in the context of PDX signaling. To confirm that this is the relevant receptor, we will measure the effect of ablating this GPCR on the PDX-mediated increase in efferocytosis and up regulation of M2 markers in RAW264.7 cells. Next, we will seek to identify the cell types that mediate the anti-endometriosis effects of multiple SPMs, including MaR1, RvD5, and PDX. To that end, we will use single- cell RNAseq (scRNAseq) to identify cell types that express SPM receptors in our mouse model of endometriosis-associated pain at a relevant timepoint. We will also measure the effect of PDX on the transcriptional profile of relevant cell types in vivo, in order to begin to understand how PDX relieves pain and reduces lesion size. Then, we will use available resources to confirm expression of these receptors in cells human lesions. When these experiments are complete, we will have identified a novel, druggable target for endometriosis therapy. As this is expected to be a GPCR, common strategies like high-throughput screening (with which we have experience) can then be used to identify pharmacophores that lack the liabilities present in PDX, itself. We will also identify pharmacodynamic markers of protectin DX activity that will allow target engagement to be measured during a therapeutic development program. Although additional target validations work (e.g. with knockout mice) will remain, these studies will provide important knowledge about the mechanism by which SPMs, including PDX, reduce endometriosis-associated pain and lesion growth. Thus, the knowledge to be gained by these studies will represent an important advance toward the development of desperately needed novel therapeutics for endometriosis.

子宫内膜异位症是一种炎症性疾病，其特征是存在子宫内膜样病变和进行性腹部或骨盆疼痛。它影响约10％的育龄妇女，约30％的患者是没有现有选择有效治疗；迫切需要新的治疗剂。我们已经开发了验证了子宫内膜异位疼痛的小鼠模型。使用该模型，我们发现特定的亲解决脂质介质（SPM）保护素DX（PDX）迅速废除子宫内膜异位相关的疼痛和收缩病变。不幸的是，稳定性和药代动力学特性差使PDX成为较差的药物铅。到必须鉴定出PDX的抗内膜异位性活性，必须鉴定出其受体。所有的SPM 亲和受体已知激活G蛋白偶联受体（GPCR）。因此，我们假设PDX作用通过GPCR并旨在识别它和表达PDX受体的细胞类型。我们将确定通过使用presto-tango系统来保护蛋白DX的机制在人类非野性GPCR的综合库中测量逮捕蛋白激活。然后我们将确定在PDX信号传导的背景下，夫妇与该GPCR的G蛋白。确认这是相关的受体，我们将衡量消融该GPCR对PDX介导的递增细胞增多症的增加和加强RAW264.7细胞中M2标记的调节。接下来，我们将寻求确定介导的细胞类型多个SPM的抗内膜异位作用，包括MAR1，RVD5和PDX。为此，我们将使用单一细胞RNASEQ（SCRNASEQ）识别在我们的小鼠模型中表达SPM受体的细胞类型在相关时间点上与子宫内膜异位相关的疼痛。我们还将测量PDX对相关细胞类型体内的转录曲线，以便开始了解PDX如何缓解疼痛和减小病变大小。然后，我们将使用可用资源来确认这些受体在细胞中的表达人体病变。这些实验完成后，我们将确定一个新型的可吸毒目标用于子宫内膜异位症治疗。因为这预计将是GPCR，诸如高通量筛查之类的常见策略（我们与之然后可以使用经验）来识别缺乏PDX本身中存在的负债的药算团。我们还将确定保护蛋白DX活动的药效学标记，这将使目标参与成为在治疗开发计划中测量。尽管其他目标验证有效（例如敲除小鼠）将保留，这些研究将提供有关机制的重要知识包括PDX在内的SPM减少子宫内膜异位症相关的疼痛和病变生长。因此，知识成为这些研究获得的将代表着迫切需要发展的重要进步用于子宫内膜异位症的新型疗法。