Mitochondria Functions Modified by Sulfotransferase 1C2

急性肾损伤（阿基）是最常见的需要住院治疗的肾脏疾病， 死亡率很高。急性肾损伤尚无可靠的治疗方法。背景下 缺血或缺氧损伤，线粒体结构早期改变损害细胞能量，激活细胞 死亡途径，增加氧自由基的产生，并可能影响肾血流动力学。不同 天然或实验诱导的对缺血性损伤的抗性的模型可以帮助鉴定生物化学， 细胞和生理事件的损伤过程，并提供了潜在的目标， 干预我们先前的工作揭示了磺基转移酶1C 2（SULT 1C 2）在细胞凋亡中的独特的、意想不到的作用。 改变线粒体生理学以提供针对缺血性损伤的保护。由于我们发现SULT 1C 2是 在从缺血预处理的肾脏分离的线粒体的蛋白质组学筛选中高度上调，我们 测试了用携带SULT 1C 2的质粒转导的肾脏是否对缺血预处理具有抗性。 该提案的目的是描述SULT 1C 2对改变的细胞增殖的贡献程度。 代谢导致缺血预处理状态。我们将检验线粒体 缺血预处理引起的适应部分是由于磺基转移酶1C 2直接作用于 线粒体功能通过改变线粒体膜中的胆固醇硫酸盐水平来实现。 在这些研究中，我们将利用两种不同的阿基抗性模型：1）褐飞虱的遗传模型， Norway大鼠和Brown Norway衍生的大鼠经济品系，以及2）实验诱导的 缺血预适应具体目标1中的研究将检验磺基转移酶1C 2 由于膜脂质组织的变化而改变线粒体呼吸蛋白组成和生理学。 这些实验将利用无标记定量质谱的蛋白质组学方法来鉴定 不同抗性模型的生化相似性。具体目标2将检验以下假设： 磺基转移酶1C 2需要线粒体受体将胆固醇转化为胆固醇硫酸盐。目标3 将检验磺基转移酶1C 2的抑制或磺基转移酶1C 2的下调 显著减弱对缺血再灌注损伤的细胞保护。这些研究将调查后- 缺血性线粒体呼吸能力、线粒体极化、肾血流动力学和肾功能 保护总的来说，拟议的研究将有助于了解细胞保护策略， 确定潜在的治疗靶点，以管理阿基的严重程度。

Acute kidney injury (AKI) is the most common renal disease requiring hospitalization and is associated with significant mortality. There remains no reliable treatment modality for acute kidney injury. In the setting of ischemia or hypoxic injury, early alterations in mitochondrial structure impair cellular energetics, activate cell death pathways, increase oxygen free radical generation and may influence renal hemodynamics. Different models of naturally or experimentally induced resistance to ischemic injury may help to identify biochemical, cellular and physiological events underlying the injury process and provide potential targets for therapeutic intervention. Our prior work uncovered a unique, unanticipated role for sulfotransferase 1C2 (SULT1C2) in changing mitochondria physiology to confer protection against ischemic injury. Since we found SULT1C2 is highly up-regulated in proteomic screens of mitochondria isolated from ischemia-preconditioned kidneys, we tested whether kidneys transduced with plasmids bearing SULT1C2 are resistant to ischemia preconditioning. The goal of this proposal is to delineate the extent of the contribution that SULT1C2 makes to altered cell metabolism resulting in an ischemia preconditioned state. We will test the hypothesis that the mitochondria adaptation due to ischemia preconditioning is due in part to direct action of sulfotransferase 1C2 on mitochondria function brought about by changing cholesterol sulfate levels in mitochondria membranes. In these studies, we will utilize two different models of resistance to AKI; 1) a genetic model of the Brown Norway rat and Brown Norway derived consomic strains of rats, and 2) a model of experimentally induced ischemic preconditioning. Studies in specific objective 1 will test the hypothesis that sulfotransferase 1C2 changes mitochondria respirome composition and physiology due to changes in membrane lipid organization. These experiments will utilize a proteomic approach of label-free-quantitative mass spectroscopy to identify biochemical similarities in different models of resistance. Specific aim 2 will test the hypothesis that sulfotransferase 1C2 requires mitochondria receptors to convert cholesterol to cholesterol sulfate. Lastly aim 3 will test the hypothesis that inhibition of sulfotransferase 1C2 or down-regulation of sulfotransferase 1C2 markedly attenuates cellular protection against ischemia reperfusion injury. These studies will investigate post- ischemic mitochondria respiratory capacity, mitochondrial polarization, renal hemodynamics and renal function protection. Overall, the proposed studies will help provide an understanding of cytoprotective strategies and identify potential therapeutic targets to manage the severity of AKI.