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

• 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
• 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receptor expression; screening; side effect; single-cell RNA sequencing; success; therapeutic development; tool; unpublished works

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性疾病，其特征是存在子宫内膜样病变， 进行性腹部或盆腔疼痛。它影响约10%的育龄妇女，约30%的患者 现有的选择不能有效治疗;迫切需要新的治疗方法。我们已经开发并 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- 分解脂质介质（SPM）保护素DX（PDX）可迅速消除糖尿病相关疼痛， 病变不幸的是，差的稳定性和药代动力学特性使得PDX成为差的药物先导。到 利用PDX的抗子宫内膜异位症活性，必须鉴定其受体。所有SPM， 已知亲和受体激活G蛋白偶联受体（GPCR）。因此，我们假设PDX作为 通过GPCR，目的是鉴定它和表达PDX受体的细胞类型。 我们将通过使用PRESTO-Tango系统来识别保护DX作用机制， 测量人类非嗅觉GPCR的综合文库中的抑制蛋白激活。然后我们将确定 在PDX信号传导背景下与该GPCR偶联的G蛋白。为了确认这与 受体，我们将测量消融这种GPCR对PDX介导的红细胞增多的影响， RAW264.7细胞中M2标志物的上调。接下来，我们将试图确定介导细胞类型。 多种SPM的抗子宫内膜异位症作用，包括MaR 1，RvD 5和PDX。为此，我们将使用单- 细胞RNAseq（scRNAseq），以鉴定在我们的小鼠模型中表达SPM受体的细胞类型。 在相关时间点的椎间盘突出相关疼痛。我们还将测量PDX对 为了开始了解PDX如何缓解疼痛， 减小病变尺寸。然后，我们将利用现有的资源来确认这些受体在细胞中的表达 人体病变 当这些实验完成后，我们将确定一个新的，药物治疗子宫内膜异位症的目标 疗法由于这预计将是一个GPCR，常见的策略，如高通量筛选（与我们 具有经验），然后可用于鉴定缺乏PDX本身中存在的负债的药效团。 我们还将确定protectin DX活性的药效学标志物，这将允许靶向参与， 在治疗开发项目中测量。尽管额外的目标验证也有效（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）可减少乳腺癌相关疼痛和病变生长。所以，知识是 这些研究所获得的成果将代表着对迫切需要的发展的重要进展。 子宫内膜异位症的新疗法。