The formation of kidney local microenvironment after acute kidney injury.

急性肾损伤后肾脏局部微环境的形成

• 批准号: 10195384
• 负责人: Dong Zhou
• 金额: $ 12.3万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10195384
• 关键词: 3-Dimensional; Acute Renal Failure with Renal Papillary Necrosis; Affect; Animal Model; Attention; Binding; Biological; Cell Communication; Cell Proliferation; Cells; Cessation of life; Chronic Kidney Failure; Complex; Creatinine; Data; Deterioration; Disease; Environment; Event; Extracellular Domain; Extracellular Matrix; Extracellular Matrix Proteins; Fibroblasts; Future; Glues; Glycoproteins; Growth Factor; Hospitalization; Immunoprecipitation; In Vitro; Injury; Injury to Kidney; Kidney; Knowledge; Ligands; Mediating; Mentored Research Scientist Development Award; Modeling; Morbidity - disease rate; Mus; Output; Pathogenesis; Process; Proteins; Proteomics; Regimen; Regulation; Renal function; Renal tubule structure; Reporting; Research; Research Personnel; Resources; Risk; Role; SHH gene; Serum; Structure; System; Techniques; Testing; Time; Tubular formation; Urine; Vertebral column; base; cost; design; experience; in vivo; injured; injury and repair; kidney repair; knock-down; mortality; new therapeutic target; novel; novel therapeutic intervention; novel therapeutics; overexpression; prevent; recruit; repaired; scaffold; tumorigenesis

PROJECT SUMMARY/ABSTRACT Acute kidney injury (AKI) is characterized by abrupt deterioration in kidney function, manifested by an increase in serum creatinine level, with or without a reduction in the amount of urine output. In the US, there was close to one million AKI hospitalizations in 2000, and this number quadrupled by 2014. The morbidity, mortality, and costs of AKI are far greater than formerly acknowledged. Regardless of the initial causes of AKI, renal tubules are considered to be the epicenter of damage. Little attention is paid to the changes of the renal local microenvironment in AKI, although the concept of a ‘microenvironment’ has already shaped our understanding of the pathogenesis of various diseases. The kidney microenvironment in AKI is comprised of different cellular components such as injured tubular cells, activated fibroblasts, extracellular matrix (ECM), and a variety of secreted factors. In general, after AKI, renal tubules undergo a repair process of dedifferentiation. During this process, ECM is an indispensable component in organizing a favorable microenvironment to promote tubule repopulation. To further understand the role of ECM in the formation of the kidney microenvironment, we constructed an ischemic kidney injury model at 10 different time points, from day 0 to 10. By using proteomics, we found that extracellular matrix protein 1 (ECM1) was the earliest activated matrix protein after ischemic AKI. We then experimentally confirmed that ECM1 was induced rapidly, as early as 4-8h after AKI, and it predominantly localizes at the foci rich in fibroblasts. In the kidney, the fibroblast is the major cellular resource of ECM synthesis. The data generated by my K01 award revealed that, in AKI, fibroblast activation is a superior early event, and it occurs far earlier than tubular cell proliferation. Meanwhile, a tubule-derived novel growth factor, Sonic Hedgehog (Shh) was secreted by renal tubules and specifically targets fibroblasts. By immunoprecipitation, we found that ECM1 can bind to the Shh ligand in vitro. Based on these findings, we hypothesized that after AKI, ECM1 directly recruits Shh to form a favorable microenvironment to promote kidney remodeling.We will test this hypothesis in two specific aims: 1) Determine the mechanistic role of ECM1 in microenvironment formation ex vivo. 2) Determine the role of the ECM1-organized microenvironment in promoting AKI repair in vivo. The data generated from this application will have wide implications in comprehending the pathogenesis of kidney repair and in designing novel therapeutic regimens for AKI treatment.

项目总结/摘要 急性肾损伤（阿基）的特征是肾功能突然恶化，表现为肾功能损害程度增加。 血清肌酐水平，伴或不伴尿量减少。在美国， 到2000年，阿基住院人数达到100万，到2014年，这一数字翻了两番。发病率、死亡率和 阿基的成本远远高于以前承认的。无论阿基的最初原因如何，肾小管 被认为是破坏的中心。对肾局部的变化很少关注， 阿基中的微环境，尽管“微环境”的概念已经形成了我们的理解 各种疾病的发病机理。阿基中的肾脏微环境由不同的细胞组成， 细胞外基质（ECM），以及多种细胞因子，如受损的肾小管细胞、活化的成纤维细胞、细胞外基质（ECM）， 分泌因子一般来说，阿基后，肾小管经历去分化的修复过程。在此 在这一过程中，ECM是组织有利的微环境以促进小管形成的不可或缺的组成部分。 再繁殖为了进一步了解ECM在肾脏微环境形成中的作用，我们 在0 - 10天的10个不同时间点构建缺血性肾损伤模型。通过使用蛋白质组学， 我们发现细胞外基质蛋白1（ECM 1）是缺血性阿基后最早活化的基质蛋白。 然后，我们实验证实，早在阿基后4-8h，ECM 1就被快速诱导， 主要定位于富含成纤维细胞的病灶。在肾脏中，成纤维细胞是主要的细胞来源 ECM的合成。我的K 01奖产生的数据显示，在阿基中，成纤维细胞活化是一个重要因素。 上级早期事件，并且其发生远早于肾小管细胞增殖。与此同时，一部由微管衍生的小说 生长因子Sonic Hedgehog（Shh）由肾小管分泌并特异性靶向成纤维细胞。通过 免疫沉淀实验发现，ECM 1在体外可与Shh配体结合。基于这些发现，我们 假设阿基后，ECM 1直接招募Shh形成有利的微环境， 我们将在两个具体目标中测试这一假设：1）确定肾脏重塑的机制， ECM 1在离体微环境形成中的作用。2)确定ECM 1组织的作用 在促进阿基修复的体内微环境中。此应用程序生成的数据将具有广泛的 理解肾修复的发病机制和设计新的治疗方案的意义 用于阿基治疗。