GENE TARGETING STRATEGIES FOR THE TREATMENT OF OSTEOGENESIS IMPERFECTA

治疗成骨不全的基因靶向策略

• 批准号: 7827085
• 负责人: David W Russell
• 金额: $ 42.9万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-25 至 2012-09-24
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7827085
• 关键词: Autologous Transplantation; Biological Assay; COL1A1 gene; COL1A2 gene; Cells; Cessation of life; Collagen; Collagen Gene; Dependovirus; Engraftment; Fracture; Funding; Gene Mutation; Gene Targeting; Genes; Genetic Polymorphism; Genetic Recombination; Hereditary Disease; Hydroxyapatites; Immunodeficient Mouse; Individual; Lentivirus Vector; Long Terminal Repeats; Measures; Mediating; Mesenchymal; Mesenchymal Stem Cells; Messenger RNA; Modeling; Mutation; Oryctolagus cuniculus; Osteoblasts; Osteogenesis; Osteogenesis Imperfecta; Patients; Peptides; Phenotype; Pluripotent Stem Cells; Polyproteins; Production; Proteins; Proviruses; Recovery; Residual state; Retroviral Vector; Site; Spumavirus; Therapeutic; Transgenes; Transplantation; United States National Institutes of Health; adeno-associated viral vector; base; bone; design; disease-causing mutation; improved; in vivo; induced pluripotent stem cell; mutant; novel therapeutics; premature; public health relevance; recombinase; research study; response; skeletal abnormality; transgene expression; treatment strategy; tricalcium phosphate; vector

DESCRIPTION (provided by applicant): This is a competitive revision application for R01 AR048328 in response to NIH Notice NOT-OD-09- 058: "NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications". Osteogenesis Imperfecta (OI) is a genetic disease caused by mutations in the type I collagen genes COL1A1 or COL1A2 that can result in major skeletal abnormalities, fractures, and premature death. We showed previously that gene targeting vectors based on adeno-associated virus (AAV) can efficiently disrupt mutant COL1A1 and COL1A2 genes in mesenchymal stem cells (MSCs) from individuals with OI, and that these MSCs then produced normal collagen and formed bone. In this R01, we received funding for three specific aims: to develop improved AAV vectors that target mutant collagen genes, to determine the effects of genetic polymorphisms on targeting, and to transplant MSCs and measure engraftment using a rabbit model. These Aims are intended to develop a therapeutic strategy consisting of isolation of a patient's MSCs, gene targeting to eliminate the mutant collagen gene, and transplantation of these autologous, targeted MSCs. While this remains a promising approach, its major shortcoming is the number of MSCs that can be obtained during ex vivo expansion. In this competitive revision application we propose to overcome this potential limitation by deriving induced pluripotent stem cells (iPSCs) from OI MSCs, which are immortal and capable of differentiating into MSCs and forming bone in vivo. We will derive iPSCs from OI MSCs, correct the collagen mutations in them by AAV-mediated gene targeting, and demonstrate these iPSCs can still form bone. PUBLIC HEALTH RELEVANCE: These experiments are meant to develop a new therapeutic paradigm, in which patient-specific pluripotent stem cells (iPSCs) are corrected by gene targeting and then returned to the patient. They will establish the feasibility of correcting genetic mutations in iPSCs and using these cells to generate bone, with major significance for the treatment of genetic diseases such as osteogenesis imperfecta.

描述（由申请人提供）：这是R 01 AR 048328的竞争性修订申请，以响应NIH通知NOT-OD-09- 058：“NIH宣布恢复法案资金可用于竞争性修订申请”。成骨不全症（OI）是一种由I型胶原基因COL 1A 1或COL 1A 2突变引起的遗传性疾病，可导致严重的骨骼异常、骨折和过早死亡。我们先前表明，基于腺相关病毒（AAV）的基因靶向载体可以有效地破坏来自OI个体的间充质干细胞（MSC）中的突变COL 1A 1和COL 1A 2基因，然后这些MSC产生正常的胶原蛋白并形成骨。在R 01中，我们获得了三个具体目标的资金：开发针对突变型胶原基因的改进的AAV载体，确定遗传多态性对靶向的影响，以及移植MSC并使用兔模型测量植入。这些目标旨在开发一种治疗策略，包括分离患者的MSC，基因靶向以消除突变型胶原基因，以及移植这些自体靶向MSC。虽然这仍然是一种有前途的方法，但其主要缺点是在离体扩增期间可以获得的MSC的数量。在这种竞争性的修订申请中，我们提出通过从OI MSC中获得诱导多能干细胞（iPSC）来克服这种潜在的限制，所述诱导多能干细胞是永生的并且能够分化成MSC并在体内形成骨。我们将从OI MSC中获得iPSC，通过AAV介导的基因靶向纠正其中的胶原突变，并证明这些iPSC仍然可以形成骨。 公共卫生关系：这些实验旨在开发一种新的治疗模式，其中患者特异性多能干细胞（iPSC）通过基因靶向进行校正，然后返回患者体内。他们将确定纠正iPSC中的基因突变并使用这些细胞生成骨骼的可行性，这对治疗遗传性疾病如骨生成障碍具有重要意义。