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receptor receptor expression screening side effect single-cell RNA sequencing success therapeutic development tool unpublished works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，进行性腹部或骨盆疼痛。它影响约 10% 的育龄妇女，约 30% 的患者患有现有方案未有效处理；迫切需要新的疗法。我们已经开发并验证了子宫内膜异位症疼痛的小鼠模型。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) 可快速消除子宫内膜异位症相关疼痛并缩小子宫内膜异位症病变。不幸的是，较差的稳定性和药代动力学特性使得 PDX 成为较差的先导药物。到要利用 PDX 的抗子宫内膜异位活性，必须鉴定其受体。所有 SPM 的高已知亲和受体可激活 G 蛋白偶联受体 (GPCR)。因此我们假设 PDX 的作用通过 GPCR 并旨在识别它和表达 PDX 受体的细胞类型。我们将通过使用 PRESTO-Tango 系统来确定 Protectin DX 的作用机制测量人类非嗅觉 GPCR 综合文库中抑制蛋白的激活。然后我们将识别在 PDX 信号传导背景下与该 GPCR 偶联的 G 蛋白。确认这是相关的受体，我们将测量消除该 GPCR 对 PDX 介导的胞吞作用增加的影响， RAW264.7 细胞中 M2 标记物的上调。接下来，我们将寻求鉴定介导该过程的细胞类型多种 SPM（包括 MaR1、RvD5 和 PDX）的抗子宫内膜异位症作用。为此，我们将使用单细胞 RNAseq (scRNAseq) 用于识别在我们的小鼠模型中表达 SPM 受体的细胞类型相关时间点子宫内膜异位症相关的疼痛。我们还将测量 PDX 对体内相关细胞类型的转录谱，以便开始了解 PDX 如何缓解疼痛和减少病灶大小。然后，我们将利用可用的资源来确认这些受体在细胞中的表达人体损伤。当这些实验完成后，我们将确定一种新的、可药物治疗子宫内膜异位症的靶点治疗。由于这预计是 GPCR，因此常见的策略如高通量筛选（我们使用它）有经验）然后可用于识别缺乏 PDX 本身存在的缺陷的药效团。我们还将确定保护素 DX 活性的药效学标记物，这将使靶标参与成为可能。在治疗开发计划期间测量。尽管额外的目标验证有效（例如，基因敲除小鼠）将继续存在，这些研究将提供有关机制的重要知识 SPM（包括 PDX）可减轻子宫内膜异位症相关的疼痛和病变生长。因此，所要掌握的知识这些研究取得的成果将代表着在开发急需的药物方面取得了重要进展子宫内膜异位症的新疗法。