Rock Inhibition as Therapy for Cerebral Cavernous Malformation

描述（由申请人提供）：脑海绵状血管畸形（CCM）是一种常见的出血性血管异常，表现为散发性和家族性常染色体显性形式。它影响了100多万美国人，使他们一生都有患中风和癫痫的风险。目前尚无预防CCM病变发生或临床进展的治疗方法。我们已经在两个CCM家族基因（Ccm 1和2）杂合的小鼠中开发了新的CCM模型，这些模型表现出重现人类疾病的CCM病变。我们的研究小组和其他研究人员已经发现RhoA/ROCK信号在杂合子Ccm 1和Ccm 2小鼠的血管通透性过高中起作用。这种高渗透性可以通过ROCK抑制剂法舒地尔或辛伐他汀逆转，辛伐他汀也会影响相同的途径。我们还提供了在小鼠病变的内皮细胞（EC）和手术切除的人CCM病变中ROCK活性增加的证据。我们假设异常的RhoA激活是CCM发病机制的最终原因，因此体内ROCK抑制将抑制病变发展和出血。鉴于每种药物的机制差异和潜在临床优势，我们建议比较法舒地尔选择性ROCK抑制作用与辛伐他汀更广泛的Rho抑制作用。我们进一步假设ROCK抑制影响相关CCM血管靶点中的分子生物标志物和相关基因表达，从而验证治疗效果。提出了三个目标来检验这些假设。在具体目标1中，我们检查了高剂量和低剂量法舒地尔或辛伐他汀对CCM病变负荷、出血和表型的影响，在生命早期和晚期开始治疗，并比较了小鼠模型CCM 1和2中的治疗效果。在特定目标2中，我们研究了高剂量和低剂量法舒地尔或辛伐他汀对通过体内MRI预筛选的已建立的小鼠CCM病变的影响，包括对病变大小、出血、表型标志物以及与ROCK和相关信号通路相关的基因的差异表达的影响。在具体目标3中，我们研究了手术切除的人CCM病变中ROCK激活的生物标志物，通过相关基因表达进行验证，作为人类治疗的潜在靶点。我们建议比较散发性和家族性背景的病变，并与三个已知的人类基因位点进行比较。这种翻译策略利用了许多机制发现和最近优化的模型，为CCM开发了一种新的可行疗法。

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.