Cyclin-dependent Kinase Inhibitor In Acute Renal Failure

我们的长期目标是预防和/或治疗急性肾功能衰竭。我们证明了激活 P21细胞周期蛋白依赖性激酶(Cdk)抑制剂基因改善顺铂所致肾功能衰竭 给药和肾脏缺血/再灌流。在这些损伤后，比较p21(+/+)和p21(-/-)小鼠， 细胞损伤显著减少，包括坏死和凋亡，肾功能下降。 失败，在p21(+/+)人群中死亡率较低。这一机制直接依赖于 P21作为细胞周期的调节因子。我们现在发现，以前使用腺病毒载体诱导p21 顺铂可完全保护体外培养的小鼠肾小管上皮细胞免受细胞毒性。 同样，用不同的药物CDK2抑制剂或组蛋白对肾细胞进行预处理 已知的诱导p21的脱乙酰酶(HDAC)抑制剂也具有保护作用。我们将这些发现扩展到 表明一种cdk2抑制药，其活性谱类似于p21，也保护肾功能。 以及顺铂所致肾损伤的体内细胞形态。我们假设CDK2抑制剂可以保护 通过抑制顺铂激活的细胞死亡途径，保护肾细胞免受顺铂的毒性作用。 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 已经开发出目标，将在体外和体内确定cdk抑制剂的保护机制，并 提供利用CDK抑制剂改善顺铂诱导的肾功能衰竭的初步步骤。 我们的第一个具体目标是确定CDK抑制剂的保护机制。我们会确认 CDK抑制剂的保护依赖于抑制CDK2的活性。我们将确定CDK2的本地化和 CDK2-p21相互作用的定位。我们将确定CDK2抑制的细胞死亡途径(S) 对于每个被CDK2抑制剂抑制的死亡途径，决定它是否被激活 顺铂。 我们的第二个具体目标是确定cdk2抑制剂的保护条件和机制。 在活体内。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 体外保护的p21膜是否也具有体内保护作用。我们将确认紫藜芦醇在 活体保护，细化其作用条件。

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 samefragment of p21 that protects in vitro also protects in vivo. We will confirm purvalanol in vivo protection and refine the conditions for its action.