Rock Inhibition as Therapy for Cerebral Cavernous Malformation

描述(申请人提供)：脑海绵状畸形(CCM)是一种常见的出血性血管畸形，以散发性和家族性常染色体显性形式表现。它影响了100多万美国人，使他们有终身中风和癫痫的风险。目前还没有预防CCM病变发生或临床进展的治疗方法。我们开发了两个CCM家族基因(Ccm1和Ccm2)杂合子的小鼠CCM新模型，这些CCM病变重现了人类疾病。我们的团队和其他人已经发现RhoA/ROCK信号与杂合子Ccm1和CCM2小鼠的血管高通透性有关。这种高渗透性可以被岩石抑制剂法舒地尔或同样影响这一途径的辛伐他汀逆转。我们提供了小鼠病变和手术切除的人CCM病变的内皮细胞(EC)中ROCK活性增加的证据。我们假设异常的RhoA激活是CCM发病的最终原因，因此体内ROCK抑制将抑制病变的发展和出血。考虑到两种药物的机制差异和潜在的临床优势，我们建议比较法舒地尔和辛伐他汀对选择性摇滚抑制和更广泛的Rho抑制的效果。我们进一步假设岩石抑制影响相关CCM血管靶点的分子生物标记物和相关基因的表达，从而验证了治疗效果。为了检验这些假设，本文提出了三个目标。在特定的目标1中，我们检测了高剂量和低剂量的法舒地尔或辛伐他汀对CCM病变负担、出血和表型的影响，并在生命早期和晚期开始治疗，并比较了在小鼠模型CCM1和2中的治疗效果。在特定的目标2中，我们检测了高剂量和低剂量的法舒地尔或辛伐他汀对通过在体MRI预先筛选的小鼠CCM病变的影响，包括对病变大小、出血、表型标志物以及ROCK相关基因和相关信号通路的差异表达的影响。在特定的目标3中，我们检测了手术切除的人类CCM病变中岩石激活的生物标记物，并通过相关基因表达进行了验证，作为人类治疗的潜在靶点。我们建议比较来自散发性和家族性背景的病变，并与三个已知的人类基因座进行比较。这一翻译策略利用了许多机械性发现和最近优化的模型，开发了一种新颖而可行的CCM治疗方法。

DESCRIPTION (provided by applicant): The cerebral cavernous malformation (CCM) is a common hemorrhagic vascular anomaly, presenting in sporadic and familial autosomal dominant forms. It affects more than a million Americans, predisposing them to a lifetime risk of stroke and epilepsy. There is currently no therapy to prevent the genesis or clinical progression of CCM lesions. We have developed novel models of CCM in mice heterozygous for two of the CCM familial genes (Ccm1 and 2), and these exhibit CCM lesions recapitulating the human disease. Our group and others have implicated RhoA/ROCK signaling in vascular hyperpermeability in heterozygous Ccm1 and Ccm2 mice. This hyperpermeability can be reversed by the ROCK inhibitor, fasudil, or by simvastatin which also impacts this same pathway. And we provided evidence of increased ROCK activity in endothelial cells (EC) of murine lesions, and in surgically resected human CCM lesions. We hypothesize that aberrant RhoA activation is the ultimate cause of CCM pathogenesis, such that in vivo ROCK inhibition will inhibit lesion development and hemorrhage. We propose to compare the effect of selective ROCK inhibition with fasudil, versus broader Rho inhibition with simvastatin, given mechanistic differences and potential clinical advantages of each drug. We further hypothesize that ROCK inhibition impacts molecular biomarkers and related gene expression in relevant CCM vascular targets, validating the therapeutic effect. Three aims are proposed to test these hypotheses. In Specific Aim 1, we examine the effect of fasudil or simvastatin at high and low doses on CCM lesion burden, hemorrhage and phenotype, with onset of treatment early and later in life, and we compare therapeutic effect in murine models CCM1 and 2. In Specific Aim 2, we examine the effect of high and low doses of fasudil or simvastatin on established murine CCM lesions pre-screened by in vivo MRI, including effects on lesion size, hemorrhage, phenotypic markers, and the differential expression of genes related to ROCK and associated signaling pathways. And in Specific Aim 3 we examine biomarkers of ROCK activation in surgically resected human CCM lesions, validated by correlative gene expression, as a potential target of therapy in man. We propose to compare lesions from sporadic and familial backgrounds, and with each of three known human gene loci. This translational strategy leverages a number of mechanistic discoveries and recently optimized models, to develop a novel and viable therapy for CCM.