Research Project

研究项目

• 批准号: 10708974
• 负责人: John C Lieske
• 金额: $ 35.44万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-23 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10708974
• 关键词: Affect; Age Months; Aging; Animal Model; Animals; Anti-Inflammatory Agents; Applications Grants; Area; Biological Markers; Biopsy; Blood; Calcium; Calcium Oxalate; Cell secretion; Cells; Clinical; Crystal Formation; Crystallization; Data; Defect; Deposition; Development; Disease; Duct (organ) structure; Effectiveness; Epithelial Cells; Event; Excretory function; Experimental Animal Model; Frequencies; Future; Grant; Growth; Human; Hydroxyapatites; Hyperoxaluria; In Vitro; Individual; Inflammatory; Kidney; Knock-out; Knockout Mice; Liquid substance; Macrophage; Maps; Mentorship; Metabolic Diseases; Morbidity - disease rate; Mus; Operative Surgical Procedures; Pain; Papillary; Pathogenesis; Pathogenicity; Patients; Persons; Phagocytes; Phagocytosis; Phenotype; Play; Population; Prevention; Process; Recurrence; Renal Tissue; Renal tubule structure; Research Personnel; Research Project Grants; Risk; Risk Factors; Role; Salts; Sampling; Scientist; Signaling Molecule; Spatial Distribution; Surface; Surgeon; Testing; Tissues; Tubular formation; Urinary Calculi; Urine; Urologic Diseases; Virulence Factors; Wild Type Mouse; Woman; Work; biobank; brushite; calcification; calcium phosphate; chemokine; comparison control; cost; cytokine; design; exosome; experience; extracellular vesicles; human tissue; hypercalciuria; inflammatory marker; innovation; interstitial; men; microvesicles; monocyte; mouse model; multidisciplinary; new therapeutic target; non-invasive monitor; novel; potential biomarker; prospective; recruit; response; skills; urinary

Project Summary/Abstract: Urinary stone disease (USD) is the 3rd most common urological disease in men and women. Prevention of USD and its associated costs and morbidity requires an understanding of early and late USD pathogenesis. Emerging evidence suggests pathophysiological interactions between intrarenal crystal nucleation, growth, and phagocytic cellular responses may play a key but unrecognized role in USD. Studies in vitro demonstrate that calcium phosphate (CaP) and calcium oxalate (CaOx) crystals induce renal tubular and phagocytic cellular secretion of cytokines, chemokines, and extracellular vesicles (EVs; exosomes and microvesicles). These biomarkers can attract blood or residential monocytes and convert monocytes into pro (M1) or anti (M2)- inflammatory macrophages (Mφ ’s ). Observations in experimental animal models and human tissues suggest that renal tissue monocytes and Mφ ’s can phagocytose and metabolize intrarenal crystals, and urinary stone formers appear to have increased medullary M1 and decreased M2 Mφ populations. In a hyperoxaluric mouse model, suppression of monocyte to M2 Mφ conversion significantly increased intrarenal CaOx deposition. Our studies also demonstrated that urinary excretion of EVs bearing inflammatory markers derived from specific segments of renal tubules were significantly lower in idiopathic calcium stone formers (ICSFs) compared to controls. Thus, multiple lines of evidence suggest that tubular and monocyte derived Mφ populations can phagocytose and degrade crystals as a crystal clearance mechanism, and defects in these clearance mechanisms could promote interstitial Randall’s plaque (RP) and/or collecting duct plug (CDP) formation. The proposed research project is designed to evaluate the role of Mφ ’s in RP and CDP formation using a novel hypercalciuric claudin-2 global knockout mouse model (over 3-24 months age) that resembles the phenotype of patients with idiopathic hypercalciuria and USD (Aim 1), and to define the frequency and spatial distribution of monocyte/ Mφ populations in carefully phenotyped ICSFs (20-70 years) with hydroxyapatite, brushite, and calcium oxalate stones plus varying amounts of RP (Aim 2). The proposed innovative study will elucidate the role of renal medullary pro-and anti-inflammatory phagocytic cells in the development of RP, CDP, and USD and whether urinary cytokines, chemokines or EVs carrying biomarkers of pro-/anti-inflammatory phagocytic cells can be used to non-invasively monitor intrarenal crystal deposition. Completion of this study will also facilitate the formation of a skilled multidisciplinary team including a promising early-stage surgeon-scientist (Dr Kevin Koo) under the mentorship of an experienced and skilled USD clinical and researcher (Dr. Lieske). The resulting preliminary data will provide evidence of the effectiveness of our team. This work will also enable submission of future detailed grant or center proposals that will extend these mechanistic studies, and has great potential to elucidate underlying pathogenic steps in USD genesisto identify novel therapeutic targets.

项目概要/摘要： 尿路结石病（USD）是男性和女性第三常见的泌尿系统疾病。预防 USD及其相关费用和发病率需要了解早期和晚期USD发病机制。 新出现的证据表明，肾内晶体成核、生长和 吞噬细胞反应可能在USD中起关键但未被认识的作用。体外研究表明， 磷酸钙（CaP）和草酸钙（CaOx）晶体诱导肾小管和吞噬细胞 细胞因子、趋化因子和细胞外囊泡（EV;外来体和微囊泡）的分泌。这些 生物标志物可以吸引血液或驻留单核细胞并将单核细胞转化为pro（M1）或anti-（M2）。 炎性巨噬细胞（Mφ’s）。在实验动物模型和人体组织中的观察表明， 肾组织单核细胞和Mφ能吞噬和代谢肾内晶体和泌尿系结石 前者髓内M1细胞数增加，M2细胞数减少。在高尿酸小鼠中， 模型中，抑制单核细胞向M2 Mφ的转化显着增加肾内CaOx沉积。我们 研究还表明，携带来自特定炎症标志物的EV的尿排泄 特发性钙结石形成者（ICSF）的肾小管节段显著低于 对照因此，多种证据表明，肾小管和单核细胞来源的Mφ群体可以 作为晶体清除机制的吞噬和降解晶体，以及这些清除中的缺陷 这些机制可促进间质性Randall斑块（RP）和/或集合管栓塞（CDP）的形成。的 建议的研究项目旨在评估的作用，Mφ的RP和CDP形成使用一种新的 高钙尿紧密连接蛋白-2整体敲除小鼠模型（超过3-24月龄），其类似于 特发性高钙尿症和USD患者（目的1），并确定频率和空间分布， 单核细胞/ Mφ群体在仔细表型ICSF（20-70岁）与羟基磷灰石，透钙磷石， 草酸钙结石加上不同量的RP（目标2）。拟议的创新研究将阐明 肾髓质促炎和抗炎吞噬细胞在RP、CDP和USD发生中的作用 以及尿细胞因子、趋化因子或EV是否携带促炎/抗炎吞噬细胞的生物标志物， 细胞可用于非侵入性地监测肾内晶体沉积。完成这项研究还将 促进形成一个熟练的多学科团队，包括一个有前途的早期外科医生-科学家（博士 Kevin Koo）在经验丰富和熟练的USD临床和研究人员（Lieske博士）的指导下进行。的 初步数据将证明我们的团队的有效性。这项工作还将使 提交未来详细的赠款或中心的建议，将扩大这些机制的研究，并已 阐明USD基因中潜在的致病步骤以确定新的治疗靶点的巨大潜力。