MOLECULAR GENETICS OF SKIN BASEMENT MEMBRANE ZONE IN EB

这一更新申请提出了广泛的研究，重点是 皮肤基底膜带（BMZ）的分子遗传学， 描绘了各种形式的大疱性表皮病的分子基础 （EB）。 拟议的研究旨在检验以下假设， BMZ结构基因中的遗传损伤是各种形式EB的基础， 精确的表型和遗传方式取决于 和特定突变的组合。 此应用程序基于solid 该项目的进展，包括：（a）克隆整个人类类型 VII胶原蛋白cDNA和相应的基因（COL 7A 1）;（B）说明 三个层粘连蛋白5基因LAMA 3、LAMB 3和 LAMC 2，以及BPAG 2和ITGB 4，连接形式的候选基因 （c）描述了EB（JEB）中COL 7A 1的60多种不同突变， EB的营养不良形式;（d）在五个不同的突变鉴定 JEB的候选基因 该提案详细说明了 集中，多学科和机构间的研究， 杰斐逊分子医学研究所的研究人员和 MGH/哈佛皮肤生物学研究中心。 五个组成项目 是高度相互依赖的。 项目一，“遗传连锁分析和 遗传性皮肤病的位置候选基因克隆”， 或排除特定基因和等位基因作为突变的候选基因， 有EB的家庭 该项目还将绘制新的角质细胞特异性 表达序列选项卡。 项目2. “克隆和表征 皮肤中表达的新基因”将提供新的基因探针， 关于EB中潜在候选基因的新基因的信息。 项目3，“EB的突变分析：基因型/表型相关性和 修订的分类，”将提供关于具体 基因/蛋白质系统中的突变在各种形式的 EB. 突变数据库的检查将允许建立 基因型/表型相关性对遗传 为有复发风险的受影响个人和家庭提供咨询 的EB。 扩展的突变分析也将形成修改的基础。 EB亚型的分类。 项目4，“联合国的职能后果” 蛋白质水平的突变”将检查表达和分泌 从受影响的个体培养的细胞中改变的蛋白质途径。 该项目还将确定对蛋白质-蛋白质至关重要的结构域 交互. 项目5. “EB基因治疗的发展”，将 专注于测试基因疗法治疗EB的方法。 该提案将突出一种新颖的，非常有前途的方法， 用于同源重组的RNA/DNA嵌合寡核苷酸。 这 多学科研究有望提供以下精确信息： 对研究成果的转化应用至关重要， 发展EB的明确分类和产前检测， 并为此提供新的基因治疗方法的基础， 一组毁灭性的皮肤病。

This renewal application proposes extensive studies focusing on the molecular genetics of the cutaneous basement membrane zone (BMZ) towards delineating the molecular basis of various forms of epidermolysis bullosa (EB). The proposed studies are designed to test the hypotheses that genetic lesions in the BMZ structural genes underlie various forms of EB, and that the precise phenotype and mode of inheritance depend on the types and combinations of specific mutations. This application is based on solid progress in this project, including: (a) cloning of the entire human type VII collagen cDNA and the corresponding gene (COL7A1); (b) elucidation of intron-exon organizations for the three laminin 5 genes, LAMA3, LAMB3, and LAMC2, as well as for BPAG2 and ITGB4, candidate genes for junctional forms of EB (JEB); (c) delineation of over 60 distinct mutations in COL7A1 in the dystrophic forms of EB; (d)identification of distinct mutations in the five candidate genes for JEB. This proposal details continuation of concentrated, multidisciplinary, and inter-institutional studies by investigators at the Jefferson Institute of Molecular Medicine and the MGH/Harvard Cutaneous Biology Research Center. The five component projects are highly interdependent. Project I, "Genetic Linkage Analysis and Positional Candidate Gene Cloning for Genodermatoses," is vital to rule in or rule out specific genes and alleles as candidate genes for mutations in families with EB. This project will also map new keratinocyte-specific expressed sequence tabs. Project 2. "Cloning and Characterization of Novel Genes Expressed in the Skin," will provide new gene probes and information about novel genes that are potential candidate genes in EB. Project 3, "Mutation Analysis in EB: Genotype/Phenotype Correlations and Revised Classification," will provide precise information on the specific mutations in the gene/protein systems that are at fault in various forms of EB. Examination of the mutation database will allow establishment of genotype/phenotype correlations with a profound impact on genetic counseling of the affected individual and families at risk for recurrence of EB. Extended mutation analysis will also form a basis for revised classification of EB subtypes. Project 4, "Functional Consequences of the Mutations at the Protein Level" will examine the expression and secretory pathways of altered proteins in cells cultured from affected individuals. This project will also identify domains critical for protein-protein interactions. Project 5. "Development of Gene Therapy for EB", will concentrate on testing gene therapy approaches towards treatment of EB. This proposal will highlight a novel, highly promising approach utilizing RNA/DNA chimeric oligonucleotides for homologous recombination. This multidisciplinary studies are expected to provide precise information of critical importance for translational applications of research towards development of definitive classification and prenatal testing of EB, as well as providing the basis for novel gene therapy approaches for this devastating group of skin diseases.