Monocyte chemoattractant Proteins and Vascular Injury

单核细胞趋化蛋白与血管损伤

• 批准号: 10732564
• 负责人: Sanjay Misra
• 金额: $ 71.02万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10732564
• 关键词: Affect; American; Angioplasty; Animals; Arteries; Arteriovenous fistula; Balloon Angioplasty; Binding; Blood Vessels; Blood flow; CCL2 gene; CCL7 gene; CCL8 gene; Cd68; Cell Adhesion Molecules; Cells; Chemotaxis; Clinical; Data; Development; Devices; Diagnosis; Disease Outcome; Doppler Ultrasound; Drug Kinetics; Electron Microscope; Encapsulated; End stage renal failure; Endothelial Cells; Endothelium; Family suidae; Fibrosis; Functional disorder; Gene Expression; Genes; Glycolates; Goals; Hemodialysis; Histologic; Hydrogels; Hyperplasia; Immune; In Vitro; Individual; Infiltration; Inflammation; Inflammatory; Intercellular adhesion molecule 1; Interleukin-8; Kidney Diseases; Legal patent; Letters; Leukocytes; Liposomes; Liquid Chromatography; Macrophage; Messenger RNA; Modulus; Molecular; Molecular Chaperones; Monocyte Chemoattractant Proteins; Oral; PECAM1 gene; Paclitaxel; Patients; Pharmaceutical Preparations; Phase I Clinical Trials; Pluronics; Procedures; Proliferating; Proteins; Recommendation; Recurrence; Renal Replacement Therapy; Risk; Role; Safety; Scanning; Smooth Muscle Myocytes; Stenosis; T cell infiltration; Testing; Time; Tunica Adventitia; Vascular Patency; Vascular remodeling; Veins; Venous; chemokine receptor; comparison control; cost; cytokine; drug release kinetics; experimental study; fatty acid-binding proteins; immune cell infiltrate; improved; inhibitor; injured; innovation; light scattering; loss of function; migration; monocyte; monocyte chemoattractant protein 1 receptor; mortality; mouse model; nanoparticle; neutralizing antibody; novel therapeutics; protein expression; recruit; restenosis; small molecule inhibitor; tandem mass spectrometry; ultrasound; vascular injury

PROJECT SUMMARY/ABSTRACT In 2017, ~750K Americans were diagnosed with end-stage kidney disease (ESKD), which rises 3% yearly. 87% will have renal replacement therapy with hemodialysis (HD), with preferred vascular access through arteriovenous fistula (AVF). AVFs have ~62% year patency due to venous stenosis (VS) and neointimal hyperplasia (VNH) causing reduced blood flow and suboptimal HD which is treated with percutaneous transluminal angioplasty (PTA) at >$3B/yr. Monocyte and macrophage recruitment occurs to the injured vessel wall after PTA of stenotic arteriovenous fistulas (AVF) through increased expression of MCP-1 leading to VS/VNH. Bindarit is an oral selective inhibitor of MCP-1, - 2, and -3 and we encapsulated it in polylactic-co-glycolic acid (PLGA) nanoparticles embedded in a thermosensitive Pluronic F127 hydrogel (BN NP) for periadventitial delivery to the outflow vein to test in this proposal. Scanning electron microscope and dynamic light scattering were used to characterize the BN NP and control nanoparticles (NP C). Liquid chromatography with tandem mass spectrometry (LC-MS/MS) was used to study drug release kinetics. Immediately after PTA, in a murine model of AVF stenosis, BN NP or NP C was administrated to the periadventitia of outflow veins. Animals were sacrificed 3 and 21 days later for gene expression, histomorphometric, and immunohistochemical analyses. Doppler ultrasound was performed weekly. There was no difference in the size and storage modulus of BN NP compared controls. Pharmacokinetic analysis demonstrated increased drug release from BN NP when compared to controls. BN NP treated vessels had reduced MCP-1, MCP-2 and MCP-3 gene and protein levels, reduced CCR2, increased FABP4/IL8, macrophage/monocyte abundance, proinflammatory cytokines, reduced CD4 (+) cells, reduced endothelial inflammation, and venous fibrosis resulting in positive vascular remodeling and improved patency with reduced VS/VNH. There was increased peak velocity 21 days after PTA in the BN NP group. Periadventitial administration of BN NP to the outflow vein after PTA results in decreased VS/VNH. Central Hypothesis. Periadventitial delivery of Bindarit NPs to AVF outflow vein after PTA decreases Mcp-1, -2, -3, CCR2 expression with increased FABP4/IL8 leading to less immune, macrophage cell infiltration with reduction in smooth muscle cells, fibrosis, and VS/VNH. We propose three specific aims: Aim 1: Determine how Bindarit NPs reduce MCP-1, -2, and -3 leading to decreased monocyte to macrophage differentiation, migration, proliferation, leukocyte chemoattraction by endothelial cells and inflammatory cytokine expression. Aim 2: Assess the role(s) of Bindarit NPs on CCR2 and FABP4/IL8 on reducing VS/VNH after PTA of stenotic AVFs. Aim 3: Ascertain the safety and efficacy of Bindarit NPs on reducing VS/VNH after PTA in pigs with CKD.

项目概要/摘要 2017 年，约 75 万美国人被诊断患有终末期肾病 (ESKD)，这一数字每年增加 3%。 87% 会 接受血液透析（HD）肾脏替代治疗，首选通过动静脉瘘的血管通路 （AVF）。由于静脉狭窄 (VS) 和新生内膜增生 (VNH) 导致 AVF 的年通畅率约为 62% 血流减少和 HD 次优，可通过经皮腔内血管成形术 (PTA) 治疗，费用 > $3B/年。 狭窄动静脉瘘 PTA 后受损血管壁发生单核细胞和巨噬细胞募集 (AVF) 通过 MCP-1 表达增加导致 VS/VNH。 Bindarit 是一种口服选择性 MCP-1 抑制剂，- 2和-3，我们将其封装在嵌入热敏材料的聚乳酸-乙醇酸共聚物（PLGA）纳米颗粒中 Pluronic F127 水凝胶 (BN NP) 用于外膜周围输送至流出静脉，以在本提案中进行测试。扫描 使用电子显微镜和动态光散射来表征 BN NP 和对照纳米颗粒 (NP C）。使用液相色谱串联质谱法 (LC-MS/MS) 研究药物释放动力学。 PTA 后，在 AVF 狭窄的小鼠模型中，立即将 BN NP 或 NP C 施用到外膜周围 的流出静脉。 3天和21天后处死动物进行基因表达、组织形态学和 免疫组织化学分析。每周进行多普勒超声检查。大小和尺寸没有区别 BN NP 的储能模量与对照相比。药代动力学分析表明药物释放增加 与对照组相比，BN NP。 BN NP 处理的血管减少了 MCP-1、MCP-2 和 MCP-3 基因和蛋白质 水平、CCR2 降低、FABP4/IL8 增加、巨噬细胞/单核细胞丰度、促炎细胞因子、 CD4 (+) 细胞减少，内皮炎症减少，静脉纤维化减少，导致积极的血管重塑 并通过减少 VS/VNH 改善通畅性。 PTA 后 21 天，BN NP 的峰值速度增加 团体。 PTA 后外膜周围给予 BN NP 至流出静脉会导致 VS/VNH 降低。 中心假设。 PTA 后外膜周围将 Bindarit NP 递送至 AVF 流出静脉可减少 Mcp-1、-2、-3、 CCR2 表达增加，FABP4/IL8 增加，导致免疫、巨噬细胞浸润减少， 平滑肌细胞、纤维化和 VS/VNH。我们提出三个具体目标： 目标 1：确定 Bindarit NP 如何减少 MCP-1、-2 和 -3，从而导致单核细胞向巨噬细胞减少 分化、迁移、增殖、内皮细胞对白细胞的化学吸引和炎症 细胞因子表达。 目标 2：评估 Bindarit NP 对 CCR2 和 FABP4/IL8 的作用，以减少狭窄 AVF PTA 后的 VS/VNH。 目标 3：确定 Bindarit NP 在减少 CKD 猪 PTA 后 VS/VNH 方面的安全性和有效性。