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.

项目摘要/摘要 急性肾脏损伤（AKI）的特征是肾功能突然定义，表现为增加 在血清肌酐水平中，有或没有减少尿液输出量。在美国，很近 在2000年为100万个AKI住院，到2014年，这一数字倍增。发病率，死亡率和 AKI的成本远远超过以前所承认的。无论AKI的最初原因如何，肾小管 被认为是损害的中心。很少关注肾脏本地的变化 尽管“微环境”的概念已经塑造了我们的理解，尽管AKI中的微环境 各种疾病的发病机理。 AKI中的肾脏微环境由不同的细胞组成 组件，例如受伤的管状细胞，活化的成纤维细胞，细胞外基质（ECM）和多种 分泌的因素。通常，在AKI之后，肾小管进行了去分化的维修过程。在此期间 过程，ECM是组织有利的微环境来促进管的必不可少的组成部分 重生。为了进一步了解ECM在肾脏微环境形成中的作用，我们 从第0天到第10天，在10个不同的时间点构建了缺血性肾脏损伤模型。通过使用蛋白质组学， 我们发现细胞外基质蛋白1（ECM1）是缺血性AKI后最早的活化基质蛋白。 然后，我们通过实验确认ECM1在AKI之后的4-8h迅速诱导，并且 主要位于富含成纤维细胞的焦点。在肾脏中，成纤维细胞是主要的细胞资源 ECM合成。我的K01奖产生的数据表明，在AKI中，成纤维细胞激活是一个 优越的早期事件，它发生的远远不如块茎细胞增殖。刻薄的是一本衍生的小说 生长因子Sonic刺猬（SHH）由肾脏块茎分泌，专门针对成纤维细胞。经过 免疫沉淀，我们发现ECM1可以在体外与SHH配体结合。基于这些发现，我们 假设AKI之后，ECM1直接招募SHH以形成一个有利的微环境 促进肾脏重塑。我们将以两个具体的目的检验这一假设：1）确定机械 ECM1在微环境形成中的作用。 2）确定ECM1组织的作用 在体内促进AKI修复方面的微环境。该应用程序产生的数据将具有较宽的 理解肾脏修复的发病机理和设计新型治疗方案的含义 用于AKI治疗。