Gene targeting with AAV vectors for the treatment of Osteogenesis Imperfecta

AAV 载体基因靶向治疗成骨不全症

• 批准号: 7904002
• 负责人: David R Deyle
• 金额: $ 12.18万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7904002
• 关键词: Achievement; Adipose tissue; Affect; Alleles; Antibodies; Base Pairing; Biological Assay; COL1A1 gene; COL1A2 gene; Cartilage; Cell Line; Cells; Cessation of life; Chromosome abnormality; Clinical; Collagen Diseases; Collagen Fibril; Collagen Gene; Collagen Type I; Deformity; Dependovirus; Development; Disease; Dominant-Negative Mutation; Exons; Fracture; Frequencies; Gene Expression Regulation; Gene Targeting; Gene Transduction Agent; Genes; Genetic Recombination; Genetic Transcription; Goals; Human; Human Cell Line; In Vitro; Individual; Inherited; Insertional Mutagenesis; Knock-out; Lead; Left; Location; Measures; Mediating; Medical Genetics; Mentors; Mesenchymal Stem Cells; Messenger RNA; Methods; Modification; Mus; Mutate; Mutation; Neonatal; Oncogene Activation; Osteogenesis Imperfecta; Patients; Polyadenylation; Process; Processed Genes; Procollagen; Production; Proteins; RNA; RNA Splicing; Recovery; Research; Research Personnel; Sequence Homologs; Single Nucleotide Polymorphism; Site; Southern Blotting; Stem cells; Structure; System; Testing; Training Programs; Universities; Variant; Viral Genes; Washington; Work; adeno-associated viral vector; arm; bone; career; cell type; effective therapy; embryonic stem cell; gene therapy; homologous recombination; improved; in vivo; insight; keratinocyte; magnetic beads; mutant; novel; novel therapeutic intervention; null mutation; prevent; programs; promoter; research study; training project; transcription termination; treatment strategy; vector

DESCRIPTION (provided by applicant): This proposal describes a five-year training program to develop an academic career in Medical Genetics and gene therapy. The program will further advance the investigator's research in the development of an AAV-mediated gene targeting strategy for the treatment of Osteogenesis Imperfecta. The investigator will be mentored by Dr. David Russell, well recognized for his achievements in developing the adeno-associated virus (AAV) vector system for gene targeting. Osteogenesis Imperfecta (01) is a group of inherited collagen disorders characterized by bone fragility with clinical manifestations varying from a mild increase in fractures to severe bone deformities and death. Mutations that cause Ol are located in the COL1A1 and COL1A2 genes. Current treatment options are limited and do not alleviate the complications seen in Ol. Most gene therapy systems involve the process of gene addition, where a promoter and a gene are delivered into a cell and integrate randomly. In contrast, gene targeting uses homologous sequences to alter the cell's endogenous genes in a site specific manner. By this method mutated genes can be either "knocked-out" or corrected. The major advantage of gene targeting is that genes can be modified at their proper location on the chromosme without disrupting cellular gene regulation. Previous work by Dr. Russell has demonstrated that an AAV-mediated gene targeting vector can target the COL1A1 gene and disrupt dominant negative mutant procollagen production. It was shown that the loss of the mutant protein improved the processing, stability and structure of the collagen fibril. The objective of this proposal is to develop an AAV gene targeting vector that will target the COL1A2 gene, reduce random integration, and target multiple individuals with Ol. The specific aims include: 1) Develop an AAV gene-targeting vector to target the COL1A2 gene in Ol; 2) Develop an improved AAV gene-targeting vector to reduce the recovery of random integrants; and 3) Determine the effects of human variation on gene targeting. The completion of this project and the training received at the University of Washington will prepare the principle investigator to be a leading researcher in gene therapy.

描述（由申请人提供）：此提案描述了一个为期五年的医学遗传学和基因治疗学术生涯的培训计划。该项目将进一步推进研究者在aav介导的治疗成骨不全的基因靶向策略方面的研究。研究者将由David Russell博士指导，他在开发用于基因靶向的腺相关病毒（AAV）载体系统方面取得了公认的成就。成骨不全症（01）是一组以骨脆性为特征的遗传性胶原蛋白疾病，临床表现从轻微骨折增加到严重骨畸形和死亡不等。导致Ol的突变位于COL1A1和COL1A2基因。目前的治疗选择是有限的，并不能减轻Ol的并发症。大多数基因治疗系统涉及基因添加过程，其中启动子和基因被传递到细胞中并随机整合。相反，基因靶向利用同源序列以特定位点的方式改变细胞的内源基因。通过这种方法，突变的基因要么被“敲除”，要么被纠正。基因靶向的主要优点是基因可以在染色体上的适当位置进行修饰，而不会破坏细胞基因的调节。Russell博士之前的研究表明，aav介导的基因靶向载体可以靶向COL1A1基因，并破坏显性阴性突变体前胶原蛋白的产生。结果表明，突变蛋白的缺失改善了胶原原纤维的加工、稳定性和结构。本提案的目的是开发一种AAV基因靶向载体，该载体将靶向COL1A2基因，减少随机整合，并靶向Ol的多个个体。具体目标包括：1)构建AAV基因靶向载体，靶向Ol中COL1A2基因；2)开发改进的AAV基因靶向载体，减少随机整合物的恢复；3)确定人类变异对基因靶向的影响。该项目的完成和在华盛顿大学接受的培训将使首席研究员成为基因治疗领域的主要研究人员。