Profilin-1's Role in Atherosclerosis

Profilin-1 在动脉粥样硬化中的作用

• 批准号: 10599835
• 负责人: Abigail Elizabeth Allen Gondringer
• 金额: $ 4.68万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10599835
• 关键词: Actin-Binding Protein; Actins; Address; Adhesions; Adopted; Agonist; Alleles; Animal Model; Apolipoprotein E; Arterial Fatty Streak; Atherosclerosis; Basic Science; Biochemical; Biological Assay; Biosensor; Blood Vessels; Cardiovascular Diseases; Cardiovascular system; Cause of Death; Cells; Cessation of life; Cytoskeleton; Development; Disease; Disease Progression; Embryo; Endothelial Cells; Endothelium; Environment; Excision; Extravasation; F-Actin; Fluorescence Resonance Energy Transfer; Foam Cells; Fostering; Genes; Genetic; Goals; Growth; Immune; In Vitro; Infiltration; Inflammatory; Intervention; Knock-out; Knockout Mice; Knowledge; Leukocytes; Ligands; Link; Measures; Mediating; Mediator of activation protein; Microfluidics; Mus; Nutrient; Pathology; Patients; Permeability; Pharmacology; Physiological; Pre-Clinical Model; Proteins; Retina; Role; Rupture; Severities; Smooth Muscle Myocytes; Testing; Therapeutic; Time; Tumor-infiltrating immune cells; United States; Vascular Endothelial Cell; Vascular Endothelium; Vascular Permeabilities; alternative treatment; angiogenesis; atherogenesis; atheroprotective; base; cancer cell; cell type; human disease; improved; in vivo; inhibitor; macrophage; migration; monocyte; mouse model; neovascularization; novel; novel therapeutics; overexpression; profilin 1; small molecule; small molecule inhibitor; therapeutic target; translational impact; translational potential

PROJECT SUMMARY Atherosclerosis is a major cause of cardiovascular-related death worldwide. Actin-binding protein Profilin-1 (Pfn1), an important regulator of actin cytoskeleton is overexpressed in atherosclerotic plaques in preclinical models as well as in human disease. Global reduction of Pfn1 by embryonic heterozygous knockout (KO) of the Pfn1 gene in mice displays diminished vascular infiltration of pro-atherogenic inflammatory cells (i.e. macrophages) and plaque size in experimentally induced atherosclerosis, suggesting that Pfn1 diminution confers partial protection against atherosclerosis. Since atherosclerosis is a disease that involves pathology of multiple cell types (vascular endothelial cells (VEC), various types of immune cells, smooth muscle cells), previous studies could not discern Pfn1’s contribution from specific cell types in atherosclerosis progression. Furthermore, lack of marketed small molecule inhibitors of Pfn1 has hampered translational efforts of targeting Pfn1 as a potential strategy to mitigate atherosclerosis development and/or progression. The goal of the proposed study is to specifically dissect the role of endothelial Pfn1 in atherosclerosis and further explore whether Pfn1 is an impactful pharmacological target to diminish atherosclerosis. To address this goal, a novel inducible VEC-specific Pfn1 KO mouse model and novel small molecule investigative compounds targeting the Pfn1-actin interaction interface will be used. Angiogenesis and vascular permeability facilitate nutrient delivery and leukocyte invasion, promoting atherogenesis, plaque growth and eventually plaque rupture. We have shown that endothelial Pfn1 is an important mediator of angiogenesis in physiological settings. We also found that Pfn1 depletion in VEC improves barrier function, a finding consistent with Pfn1’s ability to exacerbate agonist-induced microvascular hyper-permeability in vivo. Thus, we hypothesize that inhibition of vascular endothelial Pfn1 function exerts an atheroprotective effect by reducing plaque angiogenesis and vascular permeability. Aim 1A will utilize a novel EC-specific Pfn1 KO mouse to study atherosclerosis progression in Pfn1 deficient mice. In Aim 1B, in vitro endothelial perturbation of Pfn1 expression will be used to reveal the effect of Pfn1 to changes in leukocytic adhesion and extravasation in static and flow-based assays, as well as changes in endothelial contractility. Aim 2 will winnow available novel Pfn1:actin interaction inhibitors to the most effective inhibitor as determined by endothelial proliferation, migration, biochemical assays, and in vivo angiogenesis assays. The optimal inhibitor will be used in ApoE deficient mouse model as a potential therapeutic for atherosclerosis.

项目摘要 动脉粥样硬化是全球心血管相关死亡的主要原因。肌动蛋白结合蛋白Profilin-1 （Pfn 1），肌动蛋白细胞骨架的重要调节因子，在临床前动脉粥样硬化斑块中过表达 模型以及人类疾病。通过胚胎杂合敲除（KO）Pfn 1基因， Pfn 1基因在小鼠中显示减少的促动脉粥样硬化炎性细胞的血管浸润（即， 巨噬细胞）和斑块大小，表明Pfn 1减少 提供部分保护以对抗动脉粥样硬化。由于动脉粥样硬化是一种涉及 多种细胞类型（血管内皮细胞（VEC）、各种类型的免疫细胞、平滑肌细胞）， 以前的研究不能辨别动脉粥样硬化进展中特定细胞类型的Pfn 1的贡献。 此外，缺乏市售的Pfn 1小分子抑制剂阻碍了靶向的翻译努力， pfn 1作为一种潜在的缓解动脉粥样硬化发展和/或进展的策略。的目标 拟开展的研究旨在具体分析内皮细胞Pfn 1在动脉粥样硬化中的作用，并进一步探讨 pfn 1是减少动脉粥样硬化的有效药理学靶点。为了实现这一目标， VEC特异性Pfn 1 KO小鼠模型和靶向Pfn 1-肌动蛋白的新型小分子研究化合物 将使用交互界面。血管生成和血管渗透性促进营养输送， 白细胞侵入，促进动脉粥样硬化形成、斑块生长和最终斑块破裂。我们已经证明 内皮Pfn 1是生理环境中血管生成的重要介质。我们还发现Pfn 1 VEC的耗竭改善了屏障功能，这一发现与Pfn 1加剧激动剂诱导的 体内微血管通透性过高。因此，我们假设抑制血管内皮细胞Pfn 1 功能通过减少斑块血管生成和血管通透性发挥动脉粥样硬化保护作用。 目的1A利用一种新的EC特异性Pfn 1基因敲除小鼠研究Pfn 1基因缺陷小鼠动脉粥样硬化的进展 小鼠在目的1B中，Pfn 1表达的体外内皮扰动将用于揭示Pfn 1对血管内皮细胞的影响。 在静态和基于流动的测定中白细胞粘附和外渗的变化，以及 内皮收缩性目的2将筛选出最有效的新型Pfn 1：肌动蛋白相互作用抑制剂 通过内皮细胞增殖、迁移、生物化学测定和体内血管生成测定的抑制剂 测定。最佳抑制剂将在ApoE缺陷小鼠模型中用作潜在的治疗剂， 动脉粥样硬化