Cyclin-dependent Kinase Inhibitor In Acute Renal Failure

急性肾衰竭中的细胞周期蛋白依赖性激酶抑制剂

• 批准号: 7185114
• 负责人: PETER M. PRICE
• 金额: $ 22.57万
• 依托单位: UNIV OF ARKANSAS FOR MED SCIS
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-08-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7185114
• 关键词: Acute Kidney Failure; Adenovirus Vector; Apoptosis; CDKN1A gene; Cell Cycle; Cell Death; Cells; Cellular Morphology; Cessation of life; Cisplatin; Condition; Cyclin-Dependent Kinase Inhibitor; Gene Activation; Histone Deacetylase Inhibitor; In Vitro; Injury; Kidney; Kidney Failure; Knockout Mice; Mus; Necrosis; Pathway interactions; Pharmaceutical Preparations; Phosphotransferases; Physiological reperfusion; Population; Proximal Kidney Tubules; Renal function; Reperfusion Therapy; Role; Toxic effect; cytotoxicity; in vivo; inhibitor/antagonist; kidney cell; mortality; nephrotoxicity; oncoprotein p21; prevent; renal ischemia

DESCRIPTION (provided by applicant): Our long-term objective is to prevent and/or treat acute renal failure. We showed that activation of the gene for the p21 cyclin-dependent kinase (cdk) inhibitor ameliorated renal failure after cisplatin administration and renal ischemia/reperfusion. After these injuries, comparing p21(+/+) with p21(-/-) mice, there was significantly less cellular damage, including both necrosis and apoptosis, less functional kidney failure, and less mortality in the p21(+/+) population. The mechanism was directly dependent on the role of p21 as a regulator of the cell cycle. We now find that induction of p21 using an adenoviral vector before cisplatin exposure completely protected mouse kidney proximal tubule cells in vitro from cytotoxicity. Similarly, pretreatment of kidney cells with different pharmacologic cdk2 inhibitors or with a histone deacetylase (HDAC) inhibitor known to induce p21 was also protective. We extended these findings by showing that a cdk2 inhibitory drug, with a spectrum of activity similar to p21, also protected kidney function and cell morphology in vivo from cisplatin-induced renal injury. We hypothesize that cdk2 inhibitors protect kidney cells from cisplatin-induced toxicity by inhibiting cell death pathways activated by cisplatin exposure. Furthermore, we hypothesize that cdk2 inhibitors will be useful to prevent and treat acute renal failure. We have developed aims that will determine the mechanism of cdk inhibitor protection in vitro and in vivo and provide initial steps to utilize cdk inhibitors to ameliorate cisplatin-induced renal failure. Our first specific aim is to determine the mechanism of cdk inhibitor protection. We will confirm that cdk inhibitor protection is dependent on repressing cdk2 activity. We will determine cdk2 localization and the localization of cdk2-p21 interaction. We will determine the cell death pathway(s) inhibited by cdk2 inhibitors, and for each death pathway inhibited by cdk2 inhibitors, determine whether it is activated by cisplatin. Our second specific aim is to determine conditions and mechanisms of protection by cdk2 inhibitors in vivo. We will determine whether cdk2 knock-out mice are protected from cisplatin nephrotoxicity, and whether the same fragment of p21 that protects in vitro also protects in vivo. We will confirm purvalanol in vivo protection and refine the conditions for its action.

描述(申请人提供)：我们的长期目标是预防和/或治疗急性肾功能衰竭。我们发现，p21细胞周期蛋白依赖性激酶(Cdk)抑制剂基因的激活可以改善顺铂给药和肾缺血/再灌流后的肾功能衰竭。在这些损伤后，与p21(+/+)和p21(-/-)小鼠相比，p21(+/+)小鼠的细胞损伤明显较少，包括坏死和凋亡，功能性肾功能衰竭较少，死亡率较低。这一机制直接依赖于p21作为细胞周期调节因子的作用。我们现在发现，在顺铂暴露前使用腺病毒载体诱导p21完全保护体外培养的小鼠肾近端小管细胞免受细胞毒性。同样，用不同的药物CDK2抑制剂或已知可诱导p21的组蛋白脱乙酰酶(HDAC)抑制剂对肾细胞进行预处理也是有保护作用的。我们扩展了这些发现，显示了一种CDK2抑制药物，具有类似于p21的活性谱，也保护了体内的肾功能和细胞形态免受顺铂诱导的肾损伤。我们假设CDK2抑制剂通过抑制顺铂激活的细胞死亡通路来保护肾细胞免受顺铂诱导的毒性。此外，我们假设CDK2抑制剂将用于预防和治疗急性肾功能衰竭。我们开发的AIMS将在体外和体内确定CDK抑制剂的保护机制，并为利用CDK抑制剂改善顺铂诱导的肾功能衰竭提供初步步骤。我们的第一个具体目标是确定CDK抑制剂的保护机制。我们将证实CDK抑制剂的保护依赖于抑制CDK2的活性。我们将确定CDK2的定位以及CDK2-p21相互作用的定位。我们将确定CDK2抑制剂抑制的细胞死亡途径(S)，并针对每一条被CDK2抑制剂抑制的死亡途径，确定其是否被顺铂激活。我们的第二个具体目标是确定CDK2抑制剂在体内的保护条件和机制。我们将确定CDK2基因敲除小鼠是否受到顺铂肾毒性的保护，以及在体外保护作用的相同p21片段是否也在体内保护作用。我们将确认紫藜芦醇的体内保护作用，并完善其作用条件。