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.

项目摘要/摘要： 美国有435,000多名患者患有末期肾脏疾病（ESRD），预计人口将两倍 在未来的十年中。 ESRD患者，绝大多数使用长期血液透析作为肾脏替代方式的患者 治疗这些患者需要高度功能的血管访问才能获得最佳的治疗性。 血液透析弥漫性衰竭通常是从继发于新内膜增生的静脉狭窄 （VNH）。 我们的初步数据证明了VNH中IEX-1/MCP-1途径的重要作用。 AVF患者的静脉狭窄中增加了IEX-1染色。 从患有慢性肾脏疾病（CKD）的IEX-1敲​​除（KO）小鼠中恢复了显着增加 管腔血管面积，新内膜降低，面积随着细胞凋亡的显着增加，细胞的降低。 Prolity，显着，α-SMA和LY6C单核细胞染色，MCP-1显着下降 MMP-9免疫抑制剂3。 从IEX-1 KO小鼠中取出的静脉片段，暗示IEX-1会降低纤维化和炎症 功能。4。由骨乳糖酸（PLGA）组成的纳米颗粒的异常。 显着降低了第7天的IEX-1基因表达和28天后的VNH（p <0.05）。 用AVF和CKD从WT小鼠中恢复的静脉狭窄中，MCP-1显着增加 在MCP-1表达VNH的发病机理中，IEX-1的含义[11，16-22]。 血液透析放置放置后，已经观察到流出静脉处的剪切应力增加 [23-26]。 部分原因是剪切应力增加，而在上网细胞上和机械伸展 平滑肌细胞导致IEX-1基因上调，随后MCP-1增加 导致单核细胞/巨噬细胞募集的表达（卡通）。 以下内容： 特定目标1：评估内皮（EC）和血管平滑肌（VSMC）IEX-1的体内作用 CKD的小鼠AVF中的VNH形成。 IEX-1评估单核细胞和巨噬细胞的浸润，繁殖，迁移，凋亡，纤维化，管腔 AVF中MCP-1，FGF-1，TGF-B和MMP-9的直径，壁厚和表达。 假设：EC和VSMC中IEX-1表达降低的小鼠的AVF将减少静脉 狭窄形成是由于AVF MCP-1表达降低并减少了单核细胞/巨噬细胞 浸润。 特定目标2：评估MCP-1表达的静止 IEX-1删除。 腺体技术。 迁移，凋亡，纤维化，管腔直径，壁厚以及MCP-1，FGF-1，TGF-B的表达 AVF中的MMP-9。 假设：在具有CKD的小鼠的AVF中，EC和VSMC中的IEX-1表达降低，MCP-1的恢复 与AVF狭窄的Asosectsext。 特定目标3：确定钙核在预防和/或衰减带有的猪中的VNH形成中的作用 AVF和CKD。 抑制VNH。 假设：CKD的猪的Advental cartitolion向AVF的流出脉的流出静脉递送将减少 单核细胞/巨噬细胞浸润和随后的VNH，在IEX-1，MCP-1，FGF-1，FGF-1中进行下游修订 与对照组相比，TGF-β和MMP-9。