Mouse Arteries Predisposed to Neointimal Formation

小鼠动脉易于形成新内膜

• 批准号: 7576825
• 负责人: MICHAEL A. REIDY
• 金额: $ 37.75万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7576825
• 关键词: Affect; Arteries; C57BL/6 Mouse; Carotid Arteries; Cells; Data; Development; Enzymes; Epidermal Growth Factor Receptor; Event; FVB Mouse; Growth; Immigration; Injury; Knockout Mice; Lesion; Lipoproteins; Lyase; Measures; Mus; Null Lymphocytes; Phosphoric Monoester Hydrolases; Phosphotransferases; Plasma; Regulation; Signal Pathway; Signal Transduction; Small Interfering RNA; Smooth Muscle Myocytes; Sphingosine-1-Phosphate Receptor; Time; cell motility; inhibitor/antagonist; injured; kinase inhibitor; migration; prevent; receptor expression; research study; response; sphingosine 1-phosphate; sphingosine kinase; sphingosine-1-phosphate phosphatase

This proposal will examine how the sphingosine 1-phosphate (S1P) regulates smooth muscle cell migration. Our preliminary data show that SMCs of FVB mice migrate well in response to S1P and that migration is inhibited in the presence of a sphingosine kinase inhibitor (SphK). In total contrast, cells of C57BL/6 mice are not stimulated by S1P and the SphK inhibitor actually stimulates migration. We believe these differences are due to S1P receptor expression. FVB express S1P1 and S1P3 but little S1P2. In contrast C57BL/6 cells strongly express S1P2with reduced levels of S1P1 and S1P3. The first aim will measure circulating S1P levels in arteries of FVB and C57BL76 mice before and at times after injury. The activities of the S1P-kinase and S1P-phosphatase and expression of kinases, phosphatase and lyase will also be determined by PCR. S1P will also be determined in lipoprotein and in lipoprotein depleted fractions of plasma. We will also determine if plasma S1P can stimulate events in arteries and importantly affect SMC migration. The second aim will determine if loss of SphK activity influences SMC migration in injured mice arteries. These studies will use SphK null mice as well as a new inhibitor to block SphK activity. The third aim will determine the importance of specific S1P receptors for SMC to migrate. Experiments will be carried out using siRNA to S1P1, S1P2 and S1P3, and S1P2 and S1P3 null cells. The arteries of these mice will be subjected to injury and SMC migration and neointimal development measured. The final aim will determine how S1P signals and so regulates migration. These studies will ask if S1P signals via EGF receptor and if different S1P act differently. We will also determine if PKB and Rac are necessary for SMC migration and if blockade to the EGF receptor will prevent SMC migration and neointimal development in injured arteries.

该提案将研究 1-磷酸鞘氨醇 (S1P) 如何调节平滑肌细胞迁移。 我们的初步数据表明，FVB 小鼠的 SMC 对 S1P 的响应良好地迁移，并且迁移是 鞘氨醇激酶抑制剂 (SphK) 存在时会受到抑制。相比之下，C57BL/6 小鼠的细胞是 不受 S1P 刺激，而 SphK 抑制剂实际上会刺激迁移。我们相信这些差异是 由于 S1P 受体表达。 FVB表示S1P1和S1P3，但很少表示S1P2。相比之下，C57BL/6细胞 S1P2 强烈表达，S1P1 和 S1P3 水平降低。 第一个目标是测量 FVB 和 C57BL76 小鼠之前和时时动脉中的循环 S1P 水平 受伤后。 S1P-激酶和S1P-磷酸酶的活性以及激酶、磷酸酶的表达 裂解酶也将通过PCR测定。 S1P 也将在脂蛋白和脂蛋白中测定 血浆的耗尽部分。我们还将确定血浆 S1P 是否可以刺激动脉和 对SMC迁移有重要影响。第二个目标将确定 SphK 活性的丧失是否会影响 SMC 受伤小鼠动脉中的迁移。这些研究将使用 SphK 无效小鼠以及一种新的抑制剂来阻断 SphK 活动。第三个目标将确定特定 S1P 受体对于 SMC 迁移的重要性。 将使用 siRNA 对 S1P1、S1P2 和 S1P3 以及 S1P2 和 S1P3 无效细胞进行实验。这 这些小鼠的动脉将受到损伤，并测量 SMC 迁移和新内膜发育。 最终目标将确定 S1P 如何发出信号并从而调节迁移。这些研究将询问 S1P 信号是否 通过 EGF 受体，如果不同的 S1P 作用不同。我们还将确定 PKB 和 Rac 是否是必要的 SMC 迁移以及阻断 EGF 受体是否会阻止 SMC 迁移和新内膜发育 在受伤的动脉中。