Keratin Gene Targeting for the Treatment of Epidermolysis Bullosa

角蛋白基因靶向治疗大疱性表皮松解症

基本信息

批准号：
8401502
负责人：
DANIEL G MILLER
金额：
$ 30.22万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-01-15 至 2014-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8401502
关键词：
Alleles Alternative Splicing Autologous Transplantation Back Biopsy Birth Bulla Cell Culture Techniques Cell Surface Proteins Cell Transplants Cells Cessation of life Characteristics Cicatrix Clinical Trials Clone Cells Collaborations DNA Dependovirus Detection Disadvantaged Disease Dominant-Negative Mutation Epidermolysis Bullosa Epidermolysis Bullosa Simplex Epithelium Excision Exons Foundations Frequencies Future Gene Chips Gene Expression Gene Targeting Gene-Modified Genes Genetic Recombination Genetic Transcription Genome Genome Stability Genomics Growth Human Immune Immune response Infection Inherited Karyotype Keratin Lead Life Location Mediating Methods Modeling Mosaicism Mus Mutate Mutation Nude Mice Pain Patients Phenotype Population Property Proteins RNA Sequences Recurrence Reporting Signal Transduction Single-Stranded DNA Site Skin Skin Transplantation Sorting - Cell Movement Southern Blotting Stem cells Stratification Suppressor Mutations Techniques Testing Therapeutic Tissues Transcript Transplantation Undifferentiated Virginia Xenograft Model adeno-associated viral vector cellular transduction comparative genomic hybridization design effective therapy exon skipping expression vector gene therapy homologous recombination immunogenic in vivo in vivo Model keratin 5 keratinocyte medical complication mutant prevent promoter public health relevance reconstitution repository research study skin disorder treatment strategy vector

项目摘要

DESCRIPTION (provided by applicant): Epidermolysis Bullosa Simplex (EBS) is a debilitating, dominantly inherited skin blistering condition without an effective treatment. We propose to correct the phenotype of EBS-patient keratinocytes and prepare them for autologous transplantation by using a gene targeting approach. This method allows transcription from genes producing toxic proteins to be disrupted by targeted insertion of vector sequences into the mutant allele. Adeno-Associated Virus (AAV) vectors efficiently transduce primary cells in culture, and their single stranded DNA genomes have been shown to recombine with homologous chromosomal sequences. Furthermore, keratinocytes expressing abnormal keratins may be at a growth disadvantage making gene targeting strategies a plausible approach because of the growth advantage conferred to phenotypically corrected cells. The concept of growth differences of cells expressing normal and abnormal keratins is further supported by reports of revertant somatic mosaicism in patients with epidermolysis bullosa suggesting that even infrequent back mutations, or second site "suppressor" mutations can be therapeutic. Experiments described in this proposal will demonstrate the feasibility of gene targeting as a treatment approach for EBS and serve as a foundation for future studies that may lead to clinical trials for a variety of debilitating skin diseases.

描述（由申请人提供）：表皮溶解单纯胶（EBS）是一种令人衰弱的，具有有效治疗的无遗传的皮肤泡沫状况。我们建议纠正EBS患者角质形成细胞的表型，并通过使用基因靶向方法来为自体移植做准备。该方法允许从产生有毒蛋白的基因的转录受到将矢量序列靶向插入突变等位基因的破坏。腺相关病毒（AAV）载体有效地转导培养物中的原代细胞，并且它们的单个链DNA基因组已证明与同源染色体序列重新结合。此外，表达异常角蛋白的角质形成细胞可能处于生长劣势，这使得靶向基因靶向策略是合理的方法，因为具有表型型通过表型纠正的细胞的生长优势。表达正常和异常角蛋白的细胞生长差异的概念得到了表皮溶液Bullosa患者的恢复体细胞镶嵌性的报道，这表明即使是不经常的背部突变或第二个部位的“抑制剂”突变也可能具有治疗性。该提案中描述的实验将证明基因靶向作为EBS的治疗方法的可行性，并作为未来研究的基础，这可能会导致各种衰弱的皮肤疾病进行临床试验。