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： 尿路结石病（USD）是男性和女性中第三大常见的泌尿系统疾病。 USD 及其相关成本和发病率需要了解早期和晚期 USD 发病机制。 新的证据表明肾内晶体成核、生长和晶体之间的病理生理学相互作用 吞噬细胞反应可能在 USD 中发挥着关键但未被认识的作用。 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；外泌体和微泡）的分泌。 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亲（M1）或抗（M2）- 实验动物模型和人体组织中的观察结果表明炎症巨噬细胞（Mφ’s）。 肾组织单核细胞和Mφ可以吞噬和代谢肾内晶体，以及尿路结石 在高草酸尿小鼠中，前者似乎增加了髓质 M1 数量，减少了 M2 Mφ 数量。 模型中，抑制单核细胞向 M2 Mφ 转化显着增加肾内 CaOx 沉积。 研究还表明，携带源自特定炎症标记物的 EV 经尿液排泄。 与特发性钙结石形成者 (ICSF) 相比，肾小管节段明显较低 因此，多种证据表明肾小管和单核细胞衍生的 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 机制可以促进间质兰德尔斑块（RP）和/或集合管塞（CDP）的形成。 拟议的研究项目旨在使用一种新颖的方法来评估 Mφ 在 RP 和 CDP 形成中的作用 高钙尿症claudin-2全基因敲除小鼠模型（年龄超过3-24个月），类似于表型 患有特发性高钙尿症和 USD 的患者（目标 1），并确定以下情况的频率和空间分布： 使用羟基磷灰石、透钙磷石和仔细表型 ICSF（20-70 岁）中的单核细胞/ Mφ 群体 草酸钙结石加上不同量的 RP（目标 2）。 肾髓质促炎和抗炎吞噬细胞在 RP、CDP 和 USD 发展中的作用 以及尿细胞因子、趋化因子或 EV 是否携带促/抗炎吞噬细胞生物标志物 细胞可用于非侵入性监测肾内晶体沉积，这项研究的完成也将实现。 促进组建一支技术精湛的多学科团队，其中包括一位有前途的早期外科医生科学家（Dr. Kevin Koo）在经验丰富且技术精湛的美国临床和研究人员（Lieske 博士）的指导下。 由此产生的初步数据将提供我们团队有效性的证据。 提交未来详细的拨款或中心提案，以扩展这些机制研究，并已 阐明 USD 起源中潜在的致病步骤以确定新的治疗靶点具有巨大的潜力。