MOLECULAR GENETICS OF SKIN BASEMENT MEMBRANE ZONE IN EB

这份续签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 不同类型大疱性表皮松解症的分子基础 (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， 确切的表型和遗传模式取决于类型 以及特定突变的组合。此应用程序基于Solid 这一项目的进展，包括：(A)克隆整个人类类型 Vii胶原基因及其相应基因(COL7A1)；(B)阐明 三个层粘连蛋白5基因LAMA3、LAMB3和LAMB3的内含子-外显子组织 LAMC2，以及BPAG2和ITGB4，连接形式的候选基因 EB(Jeb)；(C)描绘了COL7A1中60多个不同的突变 营养不良的EB；(D)确定五种不同的突变 杰布的候选基因。这份提案详细说明了继续 集中的、多学科的和跨机构的研究 杰斐逊分子医学研究所的研究人员 麻省理工学院/哈佛皮肤生物学研究中心。五个组成部分的项目 是高度相互依存的。项目I，“遗传连锁分析和 基因皮肤病的位置候选基因克隆 或排除特定的基因和等位基因作为突变的候选基因 患有EB的家庭。该项目还将绘制新的角质形成细胞特异性图谱 表示序列选项卡。项目2.“克隆和鉴定 在皮肤中表达的新基因“将提供新的基因探针和 关于EB中潜在候选基因的新基因的信息。 项目3，“EB中的突变分析：基因型/表型相关性和 修订后的分类“将提供关于具体情况的准确信息 基因/蛋白质系统中的突变，这些突变导致各种形式的 EB.对突变数据库的检查将允许建立 基因/表型相关性对遗传有深远影响 对有复发风险的受影响个人和家庭进行咨询 关于EB的。扩展的突变分析也将成为修订的基础 EB亚型的分类。项目4，“法律的功能后果” 在蛋白质水平上的突变“将检查基因的表达和分泌 从受影响个体培养的细胞中蛋白质改变的途径。 该项目还将确定蛋白质-蛋白质的关键结构域 互动。项目5.《EB基因治疗的发展》，Will 集中精力测试EB的基因治疗方法。 这项提议将突出一种新颖的、非常有希望的方法，利用 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准确的信息 研究成果的翻译应用至关重要 EB、AS明确分类和产前检测的研究进展 并为新的基因治疗方法提供了基础 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.