Nucleophosmin Centered Diagnostics and Treatment of Ischemic Acute Kidney Injury

项目总结/摘要 我们发现，NPM 1是一种必需的Bax分子伴侣，它促进了受调控的细胞死亡和急性肾损伤。 (AKI)在缺血的人类肾脏中。NPM 1的结构-功能关系（SAR）及其相互作用位点 NPM/Bax的毒性是未知的，这限制了预防肾细胞中NPM/Bax毒性的新药的开发， 容易受到缺血性损伤。我们的综合性，跨学科的战略结合细胞生物学与结构 和药物化学，以确定使NPM对肾细胞有毒的特征并定位NPM 负责结合Bax的结构域。利用蛋白质结构分析和X射线晶体学， 和交联，我们将优化一种新的Bax肽，用于预防和治疗高危缺血性阿基。 心脏手术病人在三个目标中，我们将：（1）描述导致监管的NPM结构特征 肾细胞死亡;（2）优化药物设计以抑制NPM：Bax相互作用，以及（3）选择有效的治疗方法 预防NPM：体外Bax毒性和体内阿基。NPM在缺血性损伤中的因果作用的先前验证 人类肾脏及其在不同物种间缺血性细胞死亡过程中的保守调节 增加将我们的新疗法转化为临床的可能性。我们的科学团队拥有 在调节性细胞死亡、临床和实验性阿基、肽和分析设计、X射线晶体学 和质谱来推进我们对阿基期间缺血性细胞死亡的机制理解， 用于临床前研究的靶点和新药，并可能改变阿基患者的护理标准。

Project Summary/Abstract We discovered that NPM1, a required Bax chaperone, promotes regulated cell death and acute kidney injury (AKI) in the ischemic human kidney. The structure-function relationships (SAR) of NPM1 and its site of interaction with Bax are unknown, limiting the development of novel drugs to prevent NPM/Bax toxicity in kidney cells that are vulnerable to ischemic injury. Our integrative, cross disciplinary strategy combines cell biology with structural and medicinal chemistry to identify the features that render NPM toxic to kidney cells and localize the NPM domain responsible for binding Bax. Using protein structural analysis with x-ray crystallography, photo-labeling and cross-linking, we will optimize a novel Bax peptide for preventing and treating ischemic AKI in high risk cardiac surgery patients. In three aims, we will: (1) Characterize NPM structural features that cause regulated renal cell death; (2) Optimize drug design to inhibit NPM:Bax interaction, and (3) Select an effective therapeutic to prevent NPM:Bax toxicity in vitro and AKI in vivo. Prior validation of NPM’s causal role in ischemic injury in the human kidney and its conserved regulation during ischemic cell death across divergent species substantially increase the likelihood of translating our novel therapeutics to the clinic. Our scientific team has the combined expertise in regulated cell death, clinical and experimental AKI, peptide and assay design, x-ray crystallography, and mass spectrometry to advance our mechanistic understanding of ischemic cell death during AKI, identify targets and novel drugs for pre-clinical study, and potentially change the standard-of care for AKI patients.