Mitochondria Functions Modified by Sulfotransferase 1C2

急性肾损伤(AKI)是最常见的需要住院治疗的肾脏疾病，与 显著的死亡率。目前尚无可靠的治疗急性肾损伤的方法。在环境中 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，并可能影响肾脏血流动力学。不同 自然或实验诱导的对缺血损伤的抵抗模型可能有助于识别生化， 损伤过程中的细胞和生理事件，并为治疗提供潜在的靶点 干预。我们先前的工作发现了磺基转移酶1C2(SULT1C2)在 改变线粒体生理学以提供对缺血损伤的保护。因为我们发现SULT1C2是 从缺血预适应的肾脏分离的线粒体在蛋白质组筛选中高度上调 检测转导含有SULT1C2的质粒的肾脏是否对缺血预适应具有抵抗力。 该提案的目标是描述SULT1C2对改变的细胞的贡献程度 代谢导致缺血预适应状态。我们将检验这样的假设：线粒体 缺血预适应的部分原因是磺基转移酶1C2直接作用于 线粒体的功能是通过改变线粒体膜中的胆固醇硫酸盐水平来实现的。 在这些研究中，我们将利用两种不同的AKI抗性模型：1)棕色的遗传模型 挪威大鼠和挪威布朗大鼠衍生出大鼠的共生品系，2)实验诱导的动物模型 缺血预适应。对特定目标1的研究将检验磺基转移酶1C2的假设 由于膜脂组织的改变而改变线粒体呼吸体的组成和生理。 这些实验将利用无标记定量质谱学的蛋白质组学方法来鉴定 不同抗性模型的生化相似性。《特定目标2》将检验这一假设 磺基转移酶1C2需要线粒体受体将胆固醇转化为硫酸胆固醇。最后目标3 将检验以下假设：抑制磺基转移酶1C2或下调磺基转移酶1C2 显著减弱细胞对缺血再灌注损伤的保护作用。这些研究将调查后- 缺血线粒体呼吸量、线粒体极化、肾血流动力学和肾功能 保护。总体而言，拟议的研究将有助于了解细胞保护策略和 确定潜在的治疗靶点以管理AKI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.