Nucleophosmin Phosphorylation as a Diagnostic and Therapeutic Target for Ischemic AKI.
Nucleophosmin Phosphorylation as a Diagnostic and Therapeutic Target for Ischemic AKI.
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DOI：
10.1681/asn.2018040401
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发表时间：
2018-12
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影响因子：
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通讯作者：
Zhiyong Wang;E. Salih;C. Igwebuike;Ryan M Mulhern;R. Bonegio;A. Havasi;S. Borkan
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文献类型：
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作者：
Zhiyong Wang;E. Salih;C. Igwebuike;Ryan M Mulhern;R. Bonegio;A. Havasi;S. Borkan
背景缺血性阿基缺乏早期诊断和有效治疗的尿液标志物。差异核磷蛋白（NPM）磷酸化是一个潜在的早期标志物缺血性肾细胞损伤和治疗的目标。方法差异NPM磷酸化进行了评估，通过质谱法从小鼠和人的原代肾上皮细胞，新鲜肾组织，和尿液缺血性损伤前后收获的NPM。使用模拟应激诱导或正常NPM磷酸化的磷酸化NPM突变蛋白评估NPM的生物学行为和毒性。肽干扰NPM功能被用来探索NPM作为一个治疗target.Results的压力小时内，几乎相同的磷酸化的变化被检测到在不同的丝氨酸/苏氨酸的NPM从原代肾细胞，组织和尿液中收获的网站。一个磷酸模拟NPM蛋白，复制磷酸化压力下定位到胞质溶胶，形成单体相互作用的Bax，细胞死亡蛋白，共积累与Bax在分离的线粒体，并显着增加应激后的细胞死亡;野生型NPM或磷酸模拟NPM与正常的磷酸化配置没有。三个肾靶向肽干扰NPM在不同的功能位点显着保护细胞死亡，缺血后几个小时，将在未经处理的小鼠致命的单剂量的一种肽管理显着降低阿基的严重程度和改善survival.Conclusions这些研究结果建立磷酸化NPM作为一个潜在的早期标志物缺血性阿基，链接早期诊断与有效的治疗干预措施。
Background Ischemic AKI lacks a urinary marker for early diagnosis and an effective therapy. Differential nucleophosmin (NPM) phosphorylation is a potential early marker of ischemic renal cell injury and a therapeutic target.Methods Differential NPM phosphorylation was assessed by mass spectrometry in NPM harvested from murine and human primary renal epithelial cells, fresh kidney tissue, and urine before and after ischemic injury. The biologic behavior and toxicity of NPM was assessed using phospho-NPM mutant proteins that either mimic stress-induced or normal NPM phosphorylation. Peptides designed to interfere with NPM function were used to explore NPM as a therapeutic target.Results Within hours of stress, virtually identical phosphorylation changes were detected at distinct serine/threonine sites in NPM harvested from primary renal cells, tissue, and urine. A phosphomimic NPM protein that replicated phosphorylation under stress localized to the cytosol, formed monomers that interacted with Bax, a cell death protein, coaccumulated with Bax in isolated mitochondria, and significantly increased cell death after stress; wild-type NPM or a phosphomimic NPM with a normal phosphorylation configuration did not. Three renal targeted peptides designed to interfere with NPM at distinct functional sites significantly protected against cell death, and a single dose of one peptide administered several hours after ischemia that would be lethal in untreated mice significantly reduced AKI severity and improved survival.Conclusions These findings establish phosphorylated NPM as a potential early marker of ischemic AKI that links early diagnosis with effective therapeutic interventions.