Regulatory gene-chemokine networks in the formation of hemodialysis AVF stenosis

血液透析 AVF 狭窄形成中的调控基因-趋化因子网络

• 批准号: 9246275
• 负责人: RAJIV KUMAR
• 金额: $ 23.85万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-25 至 2020-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9246275
• 关键词: Apoptosis; Area; Arteriovenous fistula; Attenuated; Blood Vessels; Calcitriol; Caliber; Cardiovascular Diseases; Cartoons; Cell Proliferation; Cell Wall; Cells; Chronic Kidney Failure; Clinical; Clinical Trials; Data; ERG gene; End stage renal failure; Endothelial Cells; FGF1 gene; Failure; Family suidae; Fibrosis; Gelatinase B; Gene Expression; Genes; Glycolates; Goals; Hemodialysis; Histologic; Hyperplasia; Infiltration; Inflammation; Inflammatory; Knockout Mice; Matrix Metalloproteinases; Mechanics; Mediating; Molecular; Monocyte Chemoattractant Protein-1; Mus; Pathogenesis; Pathway interactions; Patient Care; Patients; Population; Proteins; Recruitment Activity; Regulation; Regulator Genes; Renal Replacement Therapy; Research; Role; Secondary to; Smooth Muscle Actin Staining Method; Smooth Muscle Myocytes; Staining method; Stains; Stenosis; Stretching; Techniques; Therapeutic; Thick; Transforming Growth Factor beta; Transforming Growth Factors; Translating; United States; Up-Regulation; Vascular Smooth Muscle; Veins; Venous; chemokine; cytokine; improved; in vivo; macrophage; migration; monocyte; nanoparticle; novel; prevent; programs; response; restoration; shear stress

PROJECT SUMMARY/ABSTRACT: More than 435,000 patients in the US have end stage renal disease (ESRD), a population expected to double in the next decade. The long-term goal of this current proposal and research program is to improve the care of patients with ESRD, the vast majority of who use long-term hemodialysis as their mode of renal replacement therapy. These patients require highly functioning vascular access for optimal therapeutic adequacy. Hemodialysis vascular access failure is frequently from venous stenosis secondary to neointimal hyperplasia (VNH). Our preliminary data demonstrate an important role for the Iex-1/Mcp-1 pathway in VNH. We show that: 1. IEX-1 staining is increased in venous stenoses removed from patients with AVF. 2. Venous stenoses in AVF removed from a Iex-1 knockout (KO) mice with chronic kidney disease (CKD) have a significant increase in lumen vessel area, decrease in neointima area with a significant increase in apoptosis, decrease in cellular proliferation, significant reduction in α-SMA and Ly6C monocyte staining, and a significant decline in MCP-1 and MMP-9 immunostaining. 3. Gene expression of Mcp-1, Fgf-1, and Tgf-β are all significantly decreased in venous segments removed from Iex-1 KO mice implying that Iex-1 decreases fibrotic and inflammatory function. 4. Adventitial delivery of nanoparticles composed of polylactic-co-glycolic acid (PLGA) with calcitriol significantly reduces Iex-1 gene expression at day 7 and VNH 28 days later (P<0.05). 5. Gene expression of Mcp-1 is significantly increased in venous stenosis removed from WT mice with AVF and CKD. These data implicate Iex-1 in the regulation of Mcp-1 expression in the pathogenesis of VNH [11, 16-22]. 6. Finally, increased shear stress at the outflow vein has been observed after hemodialysis vascular access placement [23-26]. Taken collectively, these observations support our central hypothesis: Venous stenosis in AVFs occurs in part due to an increase in shear stress upon endothelial cells and mechanical stretch upon smooth muscle cells causing up-regulation of the Iex-1 gene with subsequent increases in Mcp-1 expression which results in recruitment of monocytes/macrophages (cartoon). The specific aims are as follows: SPECIFIC AIM 1: Assess the in vivo role(s) of endothelial (EC) and vascular smooth muscle (VSMC) Iex-1 on VNH formation in AVFs of mice with CKD. We will use novel mice with EC and VSMC conditional deletion of Iex-1 to assess infiltration of monocytes and macrophages, proliferation, migration, apoptosis, fibrosis, lumen diameter, wall thickness and expression of Mcp-1, Fgf-1, Tgf-b, and Mmp-9 in the AVF. Hypothesis: AVFs of mice with decreased Iex-1 expression in ECs and VSMCs will have reduced venous stenosis formation as a result of reduced AVF Mcp-1 expression and reduced monocyte/macrophage infiltration. SPECIFIC AIM 2: Assess the effect of restoration of the Mcp-1 expression in AVFs of mice with EC and VSMC Iex-1 deletion. We will restore Mcp-1 expression in AVFs of mice with EC and VSMC deletion of Iex-1 with adenoviral technique. We will assess monocyte and macrophage infiltration, vessel wall cell proliferation, migration, apoptosis, fibrosis, lumen diameter, wall thickness and the expression of Mcp-1, Fgf-1, Tgf-b, and Mmp-9 in the AVF. Hypothesis: In AVFs of mice with CKD and reduced Iex-1 expression in EC and VSMCs, restoration of Mcp-1 expression will be associated with AVF stenosis. SPECIFIC AIM 3: Determine the role(s) of calcitriol in preventing and/or attenuating VNH formation in pigs with AVFs and CKD. We will use pigs with AVF and CKD to determine the role of adventitial delivery of calcitriol on inhibiting VNH. Hypothesis: Adventitial delivery of calcitriol to the outflow vein of AVF in pigs with CKD will have reduced monocyte/macrophage infiltration and subsequent VNH with downstream reduction in IEX-1, MCP-1, FGF-1, TGF-β, and MMP-9 compared with controls.

项目总结/摘要： 在美国，超过435，000名患者患有终末期肾病（ESRD），预计这一人群将翻一番 今后十年目前这项提案和研究计划的长期目标是改善对 ESRD患者，绝大多数使用长期血液透析作为肾脏替代模式 疗法这些患者需要高功能的血管通路以获得最佳治疗充分性。 血液透析血管通路失败通常是由于继发于新生内膜增生的静脉狭窄 （VNH）。 我们的初步数据表明，在VNH的Iex-1/Mcp-1途径的重要作用。我们表明：1。 AVF患者切除的静脉狭窄中IEX-1染色增加。2. AVF中的静脉狭窄 从患有慢性肾病（CKD）的Iex-1敲除（KO）小鼠中取出的细胞中， 管腔血管面积减少，新生内膜面积减少，细胞凋亡显著增加， 增殖，α-SMA和Ly 6C单核细胞染色显着减少，MCP-1显着下降 MMP-9免疫组化染色。3. Mcp-1、Fgf-1和TGF-β的基因表达均显著降低， 从Iex-1 KO小鼠中取出的静脉节段表明Iex-1减少了纤维化和炎症 功能4.由聚乳酸-羟基乙酸共聚物（PLGA）与骨化三醇组成的纳米粒的外膜递送 与对照组比较，Iex-1基因表达在第7天显著降低（P <0.05），VNH基因表达在第28天显著降低（P<0.05）。5.基因表达 Mcp-1在从患有AVF和CKD的WT小鼠移除的静脉狭窄中显著增加。这些数据 Iex-1参与VNH发病机制中Mcp-1表达的调节[11，16-22]。6.最后， 血液透析血管通路放置后，观察到流出静脉处的剪应力增加 [23-26]第10段。总的来说，这些观察结果支持我们的中心假设：AVF中的静脉狭窄 部分是由于内皮细胞上的剪切应力增加和 平滑肌细胞导致Iex-1基因上调，随后Mcp-1增加 表达，其导致单核细胞/巨噬细胞的募集（卡通）。具体目标如下： 如下所示： 具体目的1：评估内皮细胞（EC）和血管平滑肌细胞（VSMC）Iex-1在血管内皮细胞（EC）和血管平滑肌细胞（VSMC）中的体内作用。 CKD小鼠AVF中的VNH形成。我们将使用EC和VSMC条件性缺失的新小鼠， IEX-1评估单核细胞和巨噬细胞浸润、增殖、迁移、凋亡、纤维化、管腔 AVF的直径、壁厚和Mcp-1、Fgf-1、Tgf-b和Mmp-9的表达。 假设：在EC和VSMC中Iex-1表达降低的小鼠AVF的静脉血流量减少， 由于AVF Mcp-1表达减少和单核细胞/巨噬细胞减少导致狭窄形成 浸润 具体目的2：评估EC和VSMC小鼠AVF中Mcp-1表达的恢复作用 Iex-1缺失。我们将恢复EC和VSMC缺失Iex-1的小鼠AVF中Mcp-1的表达， 腺病毒技术。我们将评估单核细胞和巨噬细胞浸润，血管壁细胞增殖， 细胞迁移、凋亡、纤维化、管腔直径、管壁厚度以及Mcp-1、Fgf-1、Tgf-b和 AVF中的MMP-9。 假设：在患有CKD的小鼠AVF中，EC和VSMC中的Iex-1表达减少，Mcp-1的恢复 表达将与AVF狭窄相关。 具体目的3：确定骨化三醇在预防和/或减弱患有以下疾病的猪的VNH形成中的作用： AVF和CKD。我们将使用患有AVF和CKD的猪来确定骨化三醇的外膜递送对AVF和CKD的影响。 抑制VNH。 假设：在CKD猪中，骨化三醇向AVF流出静脉的外膜输送将减少 单核细胞/巨噬细胞浸润和随后的VNH，下游IEX-1，MCP-1，FGF-1， TGF-β和MMP-9与对照组相比。