Chemical, Structural and Cell-Signaling Interrogation of 15-Prostanglandin Dehydrogenase in Tissue Repair and Regeneration

15-前列腺素脱氢酶在组织修复和再生中的化学、结构和细胞信号传导研究

基本信息

批准号：
10414952
负责人：
ANDREW A PIEPER
金额：
$ 134.53万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-06-01 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10414952
关键词：
Adult Alleles Backcrossings Binding Biochemistry Biological Availability Biotechnology Bleomycin Bone Marrow Bone Marrow Transplantation Brain CXCL12 gene Cell Fraction Cells Chemicals Collection Colon Complex Cryoelectron Microscopy Cytokine Network Pathway Cytokine Signaling Data Data Set Dinoprostone Disease Disease model Drug Targeting Engineering Enzymes Exclusion Foundations Future Growth Hematopoietic Hippocampus (Brain)Individual Inflammation Injury Joints Kidney Diseases Knock-out Knockout Mice Lead Legal patent Liver LoxP-flanked allele Mediating Methods Microglia Modeling Mouse Strains Mus Neurons Neurosciences Oral Organ Oxidoreductase Partial Hepatectomy Pattern Penetration Peripheral Pharmaceutical Chemistry Pharmaceutical Preparations Pharmacology Prostaglandins Proteins Publications Pulmonary Fibrosis Recovery Reporting Research Resolution Rodent Role Signal Transduction Signaling Molecule Specificity Spleen Structure Techniques Technology Therapeutic Tissue Model Tissues Trauma Traumatic Brain Injury Ulcer Ulcerative Colitis analog base cell type cognitive function conditional knockout cytokine defined contribution design dextran sulfate sodium induced colitis improved in silico in vivo inhibitor knockout gene mRNA Expression macrophage molecular dynamics mouse model predictive test preservation prevent programs protein expression response scaffold single-cell RNA sequencing small molecule small molecule inhibitor structural biology therapeutic target three dimensional structure tissue regeneration tissue repair

项目摘要

Abstract. Prostaglandin E2 (PGE2) regulates tissue growth and repair in multiple organs. A conserved mechanism of synthesis and degradation modulates PGE2 levels in response to trauma, inflammation and disease. In particular, the enzyme 15-prostaglandin dehydrogenase (15-PGDH) is the main PGE2-degrading enzyme and therefore a key regulator of tissue repair and regeneration. 15-PGDH is an attractive drug target for diseases characterized by tissue damage. Our team successfully developed the first small molecule inhibitors of 15-PGDH with in vivo activities. In rodents, our inhibitors 1) accelerate recovery following bone marrow transplantation, 2) accelerate recovery from, or prevent, ulcerative colitis, 3) accelerate regrowth of liver tissue following partial hepatectomy, 4) ameliorate pulmonary fibrosis in a bleomycin-induced disease model, 5) enhance survival of new hippocampal neurons in adult mice, and 6) preserve cognitive function and minimize neuronal damage in mice following traumatic brain injury. Independent reports have described beneficial effects of 15-PGDH inhibition in models of renal disease and pulmonary fibrosis. We now propose a collaborative chemical, structural and cell-signaling interrogation of the role and activity of 15-PGDH. Our expertise includes medicinal chemistry, biochemistry, neuroscience, pharmacology, and structural biology. In Aim 1, we will define and exploit the structural basis for inhibition of 15-PGDH by small molecules. This aims builds on the first cryoEM structure (2.3 Å resolution) of 15-PGDH and two unrelated scaffolds of low-nM inhibitors of 15-PGDH. Proposed research aims to solve the structure of 15-PGDH in complex with new small molecule inhibitors or substrate. Computational approaches will be employed to interrogate substrate/inhibitor binding and the enzymatic mechanism. In Aim 2, we will define the cellular, protein and cytokine signaling networks that are regulated by 15-PGDH and that are engaged by 15-PGDH inhibitors to potentiate tissue regeneration and repair. The foundation of this aim includes the first demonstration of 15-PGDH activity in the brain, the identification of macrophages and microglia as major reservoirs of 15-PGDH expression in peripheral tissues and brain, respectively, and the discovery of cell and cytokine networks that respond to inhibiting 15-PGDH. We now propose to use single-cell RNA sequencing to determine the cell types that express 15-PGDH. Similar approaches will identify the cell-signaling network of induced cytokines and the cell types activated to express them. These studies will be performed in mice recovering from injury that have been treated with 15-PGDH inhibitors, along with appropriate controls. Finally, we will engineer macrophage- and microglia-targeted 15-PGDH knockouts to define the role of 15-PGDH expression in macrophages and microglia in mediating a conserved, cross-tissue response to PGE2 and 15- PGDH inhibitors. This data set will provide a foundation for future advancement of therapeutics targeting 15- PGDH and additional drug targets that modulate tissue regeneration.

抽象的。前列腺素E2（PGE2）调节多个器官的组织生长和修复。保守的合成和降解机制可调节PGE2水平，以响应创伤，注射和疾病。特别是，酶15-蛋白酶脱氢酶（15-PGDH）是主要的PGE2降解酶，因此是组织修复和再生的关键调节剂。 15-PGDH是一个有吸引力的药物目标对于以组织损伤为特征的疾病。我们的团队成功开发了第一个小分子具有体内活性的15-PGDH的抑制剂。在啮齿动物中，我们的抑制剂1）加速骨骼后的恢复骨髓移植，2）加速或预防溃疡性结肠炎，3）加速改革部分肝切开术后肝组织，4）在博来霉素诱导的疾病中改善肺纤维化模型，5）增强成年小鼠新海马神经元的生存，6）保留认知功能和创伤性脑损伤后，使小鼠的神经元损伤最小化。独立报告描述了 15-PGDH抑制在肾脏疾病和肺纤维化模型中的有益作用。现在，我们提出了对角色和活性的协作化学，结构和细胞信号询问 15-PGDH。我们的专业知识包括药物化学，生物化学，神经科学，药理学和结构生物学。在AIM 1中，我们将定义和探索抑制15-PGDH的结构基础分子。这是基于第一个冷冻结构（2.3Å分辨率）的15-PGDH和两个无关的 15-PGDH的低NM抑制剂的支架。拟议的研究旨在解决15-PGDH的结构与新的小分子抑制剂或底物配合在一起。计算方法将被聘为询问底物/抑制剂结合和酶促机制。在AIM 2中，我们将定义细胞，蛋白质和细胞因子信号网络由15-PGDH调节，由15-PGDH参与潜在组织再生和修复的抑制剂。这个目标的基础包括第一个证明大脑中15-PGDH活性，巨噬细胞和小胶质细胞的鉴定为主要外周组织和大脑中15-PGDH表达的储层分别是细胞和细胞的发现响应抑制15-PGDH的细胞因子网络。我们现在建议将单细胞RNA测序用于确定表达15-PGDH的细胞类型。类似的方法将确定细胞信号网络诱导的细胞因子和激活的细胞类型以表达它们。这些研究将在小鼠中进行从损伤中恢复了15-PGDH抑制剂以及适当的对照。最后，我们将设计巨噬细胞和小胶质细胞的15-PGDH敲除，以定义15-PGDH的作用巨噬细胞中的表达和小胶质细胞中介导对PGE2和15-的组成的跨组织反应 PGDH抑制剂。该数据集将为未来针对15-的治疗疗法的进步提供基础。 PGDH和调节组织再生的其他药物靶标。