PATHOBIOLOGY OF CEREBRAL CAVERNOUS MALFORMATION

申请人提出克隆脑海绵状血管瘤的基因， 脑畸形（CCM 1）。 中风是第三大原因， 在美国死亡，但其原因知之甚少。 CCM是一种罕见的 中风，但可能有高达0.9%的人口发病率在一般 人口 对CCM的自然历史知之甚少。 这些病变 由扩张的窦状血管空间组成，由单层 上皮 CCM通常是家族性的，但在西班牙裔中更常见 美国人 在西班牙裔美国人，特别是墨西哥裔美国人中， 疾病几乎总是与奠基者效应有关，如单倍型所示 分析. 申请人建议利用这种创始人效应 来克隆基因 他们已经将CCM 1定位在染色体7q上， 在D7S657的零重组时为10.6。 他们利用创始人效应 在西班牙裔美国人中，通过连锁不平衡定位CCM1， 间隔约220，000bp，并具有基因组DNA克隆重叠群 在这个时间段内。 他们将确定基因和突变， 西班牙裔美国人，并验证这是正确的基因， 在无关的非西班牙裔受影响的突变，因为约30%的家族性 非西班牙裔kinetics中的CCM可归因于CCM1的突变。 在具体目标2中，他们将确定 在正常和非正常的家族性和散发性CCM中， 患者的病变组织。 他们可以用疾病单倍型 在基因被克隆之前。 当他们克隆了基因， 突变将给出这些突变是否作用于 细胞水平作为功能获得或功能丧失突变。 在具体目标3中，他们将研究CCM突变的突变率， 疾病发展的自然史。 他们将扩展他们的亲属关系， 确定一组基因携带者使用疾病单倍型之前， 基因被克隆或随后发生突变。 这将有助于确定 外显率 它还将允许对未受影响的基因进行前瞻性研究， 携带者使用成像来确定疾病的自然史 研究和临床检查。 在具体目标4中，他们将在非西班牙裔中确定第二个CCM基因。 患者 他们有足够的家庭物质来实现这一目标， 缺乏异质性。 在特定目标5中，他们将定位正常和疾病中的CCM 1基因， 组织来阐明CCM的发病机制。 他们将决定 携带CCM1突变的细胞实际上导致CCM病变。 他们有 建立了这些病变的集合，并已开始发展细胞 培养不同的细胞系病变，以追求这一目标。

DESCRIPTION The applicants propose to clone the gene for cerebral cavernous malformation of the brain (CCM1). Stroke is the third leading cause of death in the US but little is known of its cause. CCM is a rare cause of stroke but may have a population incidence as high as 0.9% in the general population. Little is known of the natural history of CCM. These lesions consist of dilated sinusoidal vascular spaces lined by a single layer of epithelium. CCM is often familial but is more often familial in Hispanic Americans. In Hispanic Americans, particularly Mexican Americans, the disease is nearly always related to a founder effect as shown by haplotype analysis. The applicants propose to take advantage of this founder effect to clone the gene. They have located CCM1 on chromosome 7q with a lod score of 10.6 at zero recombination for D7S657. They have used the founder effect in Hispanic Americans to locate CCM1 by linkage disequilibrium mapping to an interval of approximately 220,000 bp and have a cloned contig of genomic DNA that spans this interval. They will identify the gene and mutation in Hispanic Americans and verify that this is the correct gene by identifying mutations in unrelated non-Hispanic affecteds since about 30% of familial CCM in non-Hispanic kindreds is attributable to mutation in CCM1. In Specific Aim 2, they will determine the prevalence and spectrum of mutations in the CCM1 gene in familial and sporadic CCM in both normal and disease tissue from patients. They can do this with the disease haplotype before the gene is cloned. When they have cloned the gene, the spectrum of mutations will give some indication of whether these mutations act at the cellular level as gain of function or loss of function mutations. In Specific Aim 3, they will look at the penetrance of CCM mutations and natural history of disease development. The will extend their kindreds and ascertain a group of gene carriers using the disease haplotype before the gene is cloned or the mutation afterwards. This will allow determination of penetrance. It will also allow prospective studies on unaffected gene carriers to determine the natural history of the disease using imaging studies and clinical examination. In Specific Aim 4, they will identify a second CCM gene in non-Hispanic patients. They have sufficient family material to carry out this aim in the absence of heterogeneity. In Specific Aim 5, they will localize the CCM1 gene in normal and disease tissue to elucidate the pathogenesis of CCM. They will determine which cells carrying CCM1 mutations actually cause the CCM lesions. They have established a collection of these lesions and have begun developing cell cultures of different cell lines from lesions in order to pursue this aim.