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.

项目摘要/摘要： 尿石病（USD）是男性和女性中第三种最常见的泌尿科疾病。预防 美元及其相关成本和发病率需要了解早期和晚期的发病机理。 新兴证据表明内部晶体成核，生长和 吞噬细胞的反应可能在美元中起关键但无法识别的作用。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，趋化因子和细胞外蔬菜（EV；外泌体和微囊泡）的分泌。这些 生物标志物可以吸引血液或居民单核细胞，并将单核细胞转化为Pro（M1）或抗（M2） - 炎症巨噬细胞（Mφ）。实验动物模型和人体组织中的观察表明 那个肾组织单核细胞和Mφ可以吞噬并代谢肾上石，尿石 造型器似乎增加了髓质M1并减少M2Mφ种群。在高氧化小鼠中 模型，将单核细胞抑制至M2Mφ转化率显着增加了肾内CAOX沉积。我们的 研究还表明，具有特定特异性的炎症标记的电动汽车的极端表达 在特发性钙石材（ICSF）中 控件。这是多种证据表明，结核病和单核细胞衍生的Mφ人群可以 吞噬细胞和降解晶体作为晶体清除机制，并在这些间隙中存在缺陷 机制可以促进兰德尔的间质兰牌（RP）和/或收集管道塞（CDP）形成。这 拟议的研究项目旨在评估Mφ在RP和CDP形成中的作用 高电钙Claudin-2全局敲除小鼠模型（超过3-24个月的年龄），类似于表型 特发性高钙尿和美元的患者（AIM 1），并定义 用羟基磷灰石，刷子和 草酸钙结石加上不同量的RP（AIM 2）。拟议的创新研究将阐明 肾脏髓质和抗炎吞噬细胞在RP，CDP和USD发展中的作用 以及携带抗/抗炎吞噬性生物标志物的尿细胞因子，趋化因子或电动汽车 细胞可用于非侵入性监测内部晶体沉积。这项研究的完成也将 促进一个熟练的多学科团队的组成 凯文·库（Kevin Koo）在经验丰富且熟练的USD临床和研究人员（Lieske博士）的心态下。这 最终的初步数据将提供我们团队有效性的证据。这项工作也将启用 提交未来的详细赠款或中心建议，这些建议将扩展这些机械研究，并具有 在USD Genesisto中阐明潜在的致​​病步骤的巨大潜力确定了新的治疗靶标。