Modifying kidney injury through p53 signaling.

通过 p53 信号传导改变肾损伤。

基本信息

批准号：
8696136
负责人：
Pierre C Dagher
金额：
$ 42.19万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-04-01 至 2018-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8696136
关键词：
Acute Acute Renal Failure with Renal Papillary Necrosis Animals Apoptosis Bacteriophages Biochemical Biological Cells Chronic Kidney Failure Clinical Complement Complex Coupled Data Development Discipline Disease Enzymes Epithelial Cells Flow Cytometry Fluorescent Probes Functional disorder Genetic Glycolysis Goals Health Housekeeping Image Immune response Inflammation Inflammatory Inflammatory Response Injury Kidney Knowledge Laboratories Lead Leukocytes Life Light Literature MSH Release-Inhibiting Hormone Measures Metabolic Metabolic Pathway Metabolism Microscopy Mission Modeling Molecular Morbidity - disease rate Natural Immunity Organ Oxidative Phosphorylation Pathway interactions Patients Phenotype Physiological Physiology Play Positron-Emission Tomography Process Protein p53 Public Health Quality of life Recovery Relative (related person)Research Respiration Role Shapes Signal Transduction Small Interfering RNA Techniques Testing Therapeutic Intervention Time Tubular formation Vascular blood supply Warburg Effect Work base cytokine expectation functional disability glucose metabolism human disease improved in vivo innovation interest kidney metabolism mortality mouse model novel pifithrin preconditioning prevent public health relevance response response to injury tumor

项目摘要

DESCRIPTION (provided by applicant): There is a fundamental gap in our understanding of the mechanisms that determine functional impairment in acute kidney injury (AKI). Continued existence of this gap presents an important clinical problem because therapeutic interventions to prevent, treat or hasten recovery in AKI have not and cannot be fully realized until this gap is filled. The long term goal is to understand the cellular and molecular regulatory functions of the protein p53 in the kidney under health and disease and to develop novel and specific therapeutic interventions targeting these mechanisms. The objective of this application, which is a step toward attainment of the long term goal, is to selectively determine how p53 regulates basal kidney metabolism and modulates inflammation during AKI. The central hypothesis of this application is that p53 is an important modulator of glucose metabolism in the kidney, as well as the innate immune response during AKI. This hypothesis has been formulated on the basis of existing literature and strong preliminary data from the applicants' laboratories. The rationale fo the proposed research is that once it is determined how p53 regulates kidney metabolism and inflammatory responses during AKI, it then permits the strategic modulation of p53 as a new and innovative pharmacological approach towards preventing and treating AKI. The central hypothesis will be tested by pursuing two specific aims: 1) Delineate, by way of advanced imaging and biochemical techniques, the capacity to shift kidney glucose metabolism towards a protective glycolytic phenotype through inhibition of p53; and 2) Determine the relative roles of tubular and leukocyte p53 in regulating the inflammatory response during AKI. Under the first aim, in vivo inhibition of p53 will be accomplished by pharmacologic inhibition (pifithrin alpha), siRNA silencing techniques, and targeted genetic deletion. Shifts in glycolysis and oxidative phosphorylation at the cellular level will be determined with intravital multiphoton microscopy of the kidney in living animals by employing fluorescent probes specific for the two major metabolic pathways and global shifts in kidney metabolism will be quantified with PET scanning techniques. These studies will be complemented by techniques to measure changes in key metabolic enzymes known to be modulated by p53. The protective capacity of this metabolic shift will be examined in models of AKI. Under the second aim, similar strategies will be used to inhibit p53 in vivo. Flow cytometry coupled with advanced molecular and cell biological techniques will be used to define alterations in the inflammatory response during AKI. These studies will be complemented by chimeric mouse models to further discern the mechanisms involved. The approach is innovative, because our approach to manipulate p53 signaling in order to modify kidney metabolism and innate immunity represents a new and substantial departure from the status quo. The proposed research is significant, because it is the next step in a continuum of research that is expected to lead to the development of more specific and targeted therapeutic interventions aimed at p53 that will prevent and limit subsequent dysfunction in AKI.

描述（由申请人提供）：我们对急性肾损伤（AKI）功能损伤的决定机制的理解存在根本性差距。这种差距的持续存在提出了一个重要的临床问题，因为在这种差距得到解决之前，预防、治疗 AKI 或加速 AKI 恢复的治疗干预措施尚未也无法完全实现。充满。长期目标是了解健康和疾病状态下肾脏中蛋白质 p53 的细胞和分子调节功能，并开发针对这些机制的新颖且具体的治疗干预措施。该应用的目的是选择性地确定 p53 如何在 AKI 期间调节基础肾脏代谢和调节炎症，这是实现长期目标的一步。该应用的中心假设是 p53 是肾脏葡萄糖代谢以及 AKI 期间先天免疫反应的重要调节剂。该假设是根据现有文献和申请人实验室提供的强有力的初步数据制定的。这项研究的基本原理是，一旦确定了 p53 在 AKI 期间如何调节肾脏代谢和炎症反应，就可以对 p53 进行战略性调节，作为预防和治疗 AKI 的一种新的创新药理学方法。中心假设将通过追求两个具体目标进行检验：1）通过先进的成像和生化技术，描绘通过抑制 p53 将肾脏葡萄糖代谢转向保护性糖酵解表型的能力； 2) 确定肾小管和白细胞 p53 在调节 AKI 期间炎症反应中的相对作用。第一个目标是通过药理抑制（pifithrin alpha）、siRNA 沉默技术和靶向基因删除来实现 p53 的体内抑制。细胞水平上糖酵解和氧化磷酸化的变化将通过活体动物肾脏的活体多光子显微镜来确定，方法是使用两种主要代谢途径特异的荧光探针，并且肾脏代谢的整体变化将通过 PET 扫描技术进行量化。这些研究将得到测量已知受 p53 调节的关键代谢酶变化的技术的补充。这种代谢转变的保护能力将在 AKI 模型中进行检查。在第二个目标下，将使用类似的策略来抑制体内 p53。流式细胞术与先进的分子和细胞生物学技术相结合将用于确定 AKI 期间炎症反应的变化。这些研究将得到嵌合小鼠模型的补充，以进一步辨别所涉及的机制。该方法具有创新性，因为我们操纵 p53 信号传导以改变肾脏代谢和先天免疫的方法代表了与现状的新的实质性背离。拟议的研究意义重大，因为它是连续研究的下一步，预计将导致针对 p53 的更具体和更有针对性的治疗干预措施的开发，从而预防和限制随后的 AKI 功能障碍。