Rho Kinase Inhibition Rescues the Endothelial Cell Cerebral Cavernous Malformation Phenotype
Rho Kinase Inhibition Rescues the Endothelial Cell Cerebral Cavernous Malformation Phenotype
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DOI：
10.1074/jbc.c109.097220
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发表时间：
2010-04-16
影响因子：
4.8
通讯作者：
Johnson, Gary L.
中科院分区：
文献类型：
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作者：
Borikova, Asya L.;Dibble, Christopher F.;Johnson, Gary L.
脑海绵状血管瘤（CCM）是导致癫痫发作和中风的血管病变。导致 ccm1、-2 或 -3 三个基因之一失活的突变足以诱发血管内皮细胞缺陷，从而导致 CCM。在此，我们发现 CCM1、-2 或 -3 蛋白表达的丧失会导致 GTPase RhoA 表达显着增加。使用 RhoA 特异性生物传感器进行的活细胞成像表明，RhoA 活性增加，同时 CCM1、-2 或 -3 缺失，并且在细胞质和细胞核中 RhoA 激活特别明显，同时 CCM1 表达缺失。 RhoA 激活增加与肌球蛋白轻链 2 的 Rho 激酶依赖性磷酸化相关。从功能上讲，CCM1、-2 或 -3 的缺失会抑制内皮细胞血管样管形成和细胞外基质侵袭，这两种情况均可通过化学抑制或 Rho 激酶的短发夹 RNA 敲低来挽救。这些发现首次定义了 CCM 病理学中 CCM1、-2 和 -3 的信号网络，其中 CCM1、-2 或 -3 蛋白表达的丧失导致 RhoA 活性增加，而 Rho 激酶的激活负责内皮细胞失调。结果将 Rho 激酶定义为拯救内皮细胞免于 CCM 蛋白功能丧失的治疗靶点。
Cerebral cavernous malformations (CCM) are vascular lesions causing seizures and stroke. Mutations causing inactivation of one of three genes, ccm1, -2, or -3, are sufficient to induce vascular endothelial cell defects resulting in CCM. Herein, we show that loss of expression of the CCM1, -2, or -3 proteins causes a marked increase in expression of the GTPase RhoA. Live cell imaging with a RhoA-specific biosensor demonstrates increased RhoA activity with loss of CCM1, -2, or -3, with an especially pronounced RhoA activation in both the cytosol and the nucleus with loss of CCM1 expression. Increased RhoA activation was associated with Rho kinase-dependent phosphorylation of myosin light chain 2. Functionally, loss of CCM1, -2, or -3 inhibited endothelial cell vessel-like tube formation and extracellular matrix invasion, each of which is rescued by chemical inhibition or short hairpin RNA knockdown of Rho kinase. The findings, for the first time, define a signaling network for CCM1, -2, and -3 in CCM pathology, whereby loss of CCM1, -2, or -3 protein expression results in increased RhoA activity, with the activation of Rho kinase responsible for endothelial cell dysregulation. The results define Rho kinase as a therapeutic target to rescue endothelial cells from loss of CCM protein function.