In Vivo Gene Targeting to Treat Inherited Bone Disease

体内基因靶向治疗遗传性骨病

基本信息

批准号：
9088355
负责人：
David R Deyle
金额：
$ 34.98万
依托单位：
MAYO CLINIC ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2021-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9088355
关键词：
Address Affect Aftercare Alleles Bone Development Bone Diseases Bone Marrow Bone Regeneration COL1A2 gene Cell Cycle Kinetics Cells Cessation of life Clinical Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Collagen Collagen Diseases Collagen Fibril Collagen Gene Connective Tissue Diseases DNA DNA Sequence Alteration Defect Deformity Dependovirus Disease Dose Effectiveness Excision Fracture Frequencies Gene Targeting Generations Genetic Goals Growth Hepatocyte Human Genome Impairment Implant In Vitro Inborn Genetic Diseases Individual Inherited Investigation Knock-in Mouse Lead Mediating Methods Minerals Modeling Modification Monitor Morphology Mus Mutation Neonatal Orthopedic Procedures Oryctolagus cuniculus Osteoblasts Osteogenesis Imperfecta Patient-Focused Outcomes Patients Phenotype Pre-Clinical Model Procollagen Production Proteins Protocols documentation RNA Research Rodent Series Signal Transduction Site Stem cells Stromal Cells Structure Symptoms System Tamoxifen Terminator Codon Therapeutic Tissues Transforming Growth Factor beta Transforming Growth Factors Work adeno-associated viral vector biobank bone bone fragility bone metabolism cell type design effective therapy embryonic stem cell experimental study gene correction gene therapy genetic manipulation genome editing improved in vivo in vivo Model keratinocyte long bone mouse model mutant novel novel therapeutic intervention null mutation pre-clinical precursor cell premature public health relevance retinal rods scaffold skeletal stem cell fate stem cell therapy success symptomatic improvement targeted delivery transcriptome vector

项目摘要

DESCRIPTION (provided by applicant): Mutations in bone stem cells can impair bone metabolism and regeneration. Osteogenesis imperfect (OI) is the most common form of genetic bone disease and characterized by bone fragility with clinical manifestations varying from a mild increase in fractures to severe bone deformities and death. Individuals with OI have mutations that alter procollagen structure or production and lead to abnormal collagen fibril formation disrupting bone development and renewal. Treatment options for OI are limited and do not alleviate the complications seen in OI. Our recent work has demonstrated that AAV gene targeting vectors can disrupt the mutant collagen alleles that cause OI and that targeted cells produced normal collagen and formed bone. While these results indicate the effectiveness of genomic editing with targeting frequencies approaching 1%, an enhancement of AAV gene targeting is required to provide more robust in vivo genetic manipulations. Here we will expand upon our successes and propose three lines of investigations to develop a therapeutic approach for patients with inherited bone disease. First we will modify collagen genes using a CRISPR-AAV system that will allow for the disruption or correction of genes that cause OI in vivo. Second, we will investigate whether genetically corrected bone stem cells have a growth advantage in vivo and how TGF-beta signaling affects the extra-skeletal manifestations of bone disease. Finally we will conduct a series of experiments in a preclinical rabbit model to establish the optimal gene targeting delivery protocol. Successful completion of this proposal will lead to an improved understand of TGF-beta signaling and stem cell kinetics in bone and advance AAV gene targeting toward a clinical trial.

描述（通过应用提供）：骨干细胞中的突变会损害骨代谢和再生。成骨不完善（OI）是遗传骨骼疾病的最常见形式，其特征是骨骼脆弱性，临床表现从骨折的轻度增加到严重的骨骼畸形和死亡。 OI的个体具有改变胶原素结构或生产的突变，并导致异常的胶原原纤维形成破坏骨骼发育和更新。 OI的治疗选择有限，并且不会减轻OI中的并发症。我们最近的工作表明，靶向载体的AAV基因会破坏引起OI的突变胶原等位基因，并且靶向细胞会产生正常的胶原蛋白并形成骨头。尽管这些结果表明基因组编辑的有效性接近1％的靶向频率，但需要增强AAV基因靶向的靶向，以提供更强大的体内通用操作。在这里，我们将扩大我们的成功和提议，为遗传性骨骼疾病患者开发一种治疗方法。首先，我们将使用CRISPR-AAV系统修改胶原蛋白基因，该系统将允许破坏或校正导致体内OI的基因。其次，我们将研究遗传校正的骨干细胞在体内是否具有生长优势，以及TGF-β信号如何影响骨骼疾病的骨骼外表现。最后，我们将在临床前兔模型中进行一系列实验以建立最佳基因靶向输送方案。该提案的成功完成将导致对骨骼中TGF-β信号传导和干细胞动力学的了解，并推进针对临床试验的AAV基因。