Project 1

项目1

基本信息

批准号：
10684170
负责人：
ERIC N Olson
金额：
$ 53.29万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
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：
2015-09-15 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10684170
关键词：
Affect Aging Animal Model Animal Muscular Dystrophy Applications Grants Brain CRISPR/Cas technology Canis familiaris Cell Nucleus Cell model Cessation of life Clinic Clustered Regularly Interspaced Short Palindromic Repeats Collaborations DNA Sequence Alteration Dependovirus Disease Dose Duchenne muscular dystrophy Dystrophin Exercise Exons Funding Gene Expression Profile Gene Mutation Genes Genome Genomic approach Genomics Goals Heart Hot Spot Human Individual Inherited Injury Link Longevity Luciferases Mediating Mus Muscle Muscle Fibers Muscle Weakness Muscle function Muscle satellite cell Muscular Dystrophies Mutation Myoblasts Myocardium Nuclear Oligonucleotides Patients Proteins Reading Frames Reporter Scientist Skeletal Muscle System Technology Therapeutic Time Tissues Translating United States National Institutes of Health Viral Work boys disease-causing mutation exon skipping genome editing in vivo in vivo monitoring mouse model muscular dystrophy mouse model muscular structure mutant mutation correction non-invasive monitor novel therapeutic intervention postnatal premature prevent response single nucleus RNA-sequencing stem cells therapeutic genome editing

项目摘要

Project Summary/Abstract Duchenne muscular dystrophy (DMD) is an inherited X-linked disease caused by mutations in the gene encoding dystrophin, a protein required for muscle fiber integrity. The dystrophin gene is one of the largest human genes and consists of 79 exons. Although there are thousands of individual DMD mutations that have been identified in humans, these mutations are concentrated in hot spot regions of the dystrophin gene. DMD affects approximately 1 in 5,000 boys and is characterized by progressive severe muscle weakness and a shortened lifespan. Despite intense efforts to find cures for DMD through a variety of approaches, including myoblast transfer, viral delivery of dystrophin, and oligonucleotide-mediated exon skipping, there remains no cure for this disease. Our approach is to use CRISPR/Cas9 genomic editing to permanently correct DMD by skipping or reframing the mutant dystrophin exons in postnatal muscle tissue in vivo. We refer to this strategy as Myoediting. This genome editing approach removes the genetic mutation responsible for the disease, allowing for permanent correction of muscle structure and function. We deliver the CRISPR/Cas9 components using an adeno-associated virus-9 (AAV9) delivery system which has been shown to provide robust expression in skeletal muscle, heart and brain, the major tissues affected in DMD patients. To date, we have successfully corrected the dystrophin gene mutation in several DMD animal models having mutations in key hot spot regions of the dystrophin gene. In the previous funding period, we generated several other DMD animals models covering the remaining human hot spot regions and propose to correct these mutations using CRISPR/Cas genomic editing. Although we have made much progress using CRISPR/Cas genomic editing to correct DMD, there remains more work to be done to translate this gene editing therapy to the clinic. The efficiency of delivering the CRISPR/Cas9 components needs to be optimized and questions remain as to the durability of dystrophin expression after correction. Furthermore, since muscle fibers have hundreds of nuclei, we need to understand the occurrence of CRISPR/Cas9 genomic editing at the individual nuclear level. The long-term goal of this project remains to optimize and adapt CRISPR/Cas9-mediated genome editing to postnatal muscle and ultimately to leverage this approach to correct DMD mutations in humans. This project continues to represent a close collaboration between clinicians and basic scientists sharing the common goal of advancing a new therapeutic strategy to permanently cure DMD.

项目摘要/摘要 Duchenne肌肉营养不良症（DMD）是由基因突变引起的遗传X连锁疾病编码肌营养不良蛋白，这是肌肉纤维完整性所需的蛋白质。肌营养不良的基因是最大的基因之一人类基因，由79个外显子组成。尽管有成千上万个单独的DMD突变这些突变已在人类中鉴定出来，集中在肌营养不良基因的热点区域。 DMD 影响大约5,000名男孩中的1个，其特征是进行性严重的肌肉无力和寿命缩短。尽管努力通过各种方法来寻找DMD的方法，包括成肌细胞转移，肌营养不良蛋白的病毒递送和寡核苷酸介导的外显子跳过，没有治愈这种疾病。我们的方法是使用CRISPR/CAS9基因组编辑以永久纠正DMD 在体内跳过或重塑突变肌营养不良蛋白外显子。我们参考这个策略作为myoeding。这种基因组编辑方法消除了负责该疾病的基因突变，允许永久校正肌肉结构和功能。我们提供CRISPR/CAS9组件使用与腺相关的病毒-9（AAV9）输送系统，该系统已证明可提供强大的在骨骼肌，心脏和大脑中的表达，DMD患者受影响的主要组织。迄今为止，我们有成功纠正了在钥匙中具有突变的几种DMD动物模型中的肌营养不良蛋白基因突变肌营养不良基因的热点区域。在上一个资金期间，我们生成了其他几个DMD 涵盖其余人类热点区域的动物模型，并建议使用 CRISPR/CAS基因组编辑。尽管我们使用CRISPR/CAS基因组编辑取得了很大的进步正确的DMD，将这种基因编辑疗法转化为诊所仍有更多的工作要做。这交付CRISPR/CAS9组件的效率需要优化，并且有关校正后肌营养不良蛋白表达的耐用性。此外，由于肌肉纤维具有数百个核，因此我们需要了解在个体核水平上CRISPR/CAS9基因组编辑的发生。这该项目的长期目标仍然是优化和调整CRISPR/CAS9介导的基因组编辑产后肌肉，最终利用这种方法来纠正人类的DMD突变。这个项目继续代表临床医生与基础科学家之间的密切合作，共享共同的目标推进一种新的治疗策略以永久治愈DMD。