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

基本信息

批准号：
8315912
负责人：
David R Deyle
金额：
$ 12.16万
依托单位：
MAYO CLINIC ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-09-01 至 2013-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8315912
关键词：
Achievement Adipose tissue Affect Alleles Antibodies Base Pairing Biological Assay CD34 gene 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 Internal Ribosome Entry Site 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

项目摘要

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）：该提案描述了一项为期五年的培训计划，以发展医学遗传学和基因疗法的学术生涯。该计划将进一步推进研究者在开发AAV介导的基因靶向骨生成不完美的基因策略方面的研究。该研究人员将由戴维·罗素（David Russell）博士指导，他在开发腺相关病毒（AAV）载体系统方面取得的成就而受到认可。成骨不完美（01）是一组遗传性胶原蛋白疾病，其特征是骨骼脆弱性，其临床表现从骨折的轻度增加到严重的骨骼畸形和死亡。引起OL的突变位于COL1A1和COL1A2基因中。当前的治疗选择是有限的，不会减轻OL中的并发症。大多数基因治疗系统都涉及添加基因的过程，其中启动子和基因被传递到细胞中并随机整合。相反，基因靶向使用同源序列以特定于位点的方式改变细胞的内源基因。通过这种方法，突变的基因可以被“敲除”或校正。基因靶向的主要优点是，可以在染色体上的适当位置修改基因而不会破坏细胞基因调节。罗素博士先前的工作表明，AAV介导的靶向载体的基因可以瞄准COL1A1基因并破坏显性的负突变体产量。结果表明，突变蛋白的丧失改善了胶原原纤维的加工，稳定性和结构。该提案的目的是开发一个靶向载体的AAV基因，该基因将靶向COL1A2基因，减少随机整合并针对多个具有OL的个体。具体目的包括：1）开发一个靶向AAV基因靶向载体，以靶向OL中的Col1a2基因； 2）开发改进的AAV基因靶向载体，以减少随机整合体的恢复； 3）确定人类变异对基因靶向的影响。该项目的完成以及华盛顿大学接受的培训将使主要研究人员成为基因疗法的主要研究人员。