Selectable Editing of Gene Defects Affecting Solid Tissu

影响实体组织的基因缺陷的选择性编辑

• 批准号: 6735353
• 负责人: FRANK TAGLIAFERRI
• 金额: $ 10万
• 依托单位: TAPESTRY PHARMACEUTICALS, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6735353
• 关键词: DNA repair; animal tissue; cell line; dogs; flow cytometry; gene mutation; gene therapy; genetic manipulation; laboratory mouse; method development; muscle cells; muscular dystrophy; plasmids; sickle cell anemia; tissue /cell culture; transfection /expression vector

DESCRIPTION (provided by applicant): Inherited diseases, occurring in solid tissues in the body, are often caused by simple recessive mutations, derived from two carrier parents with a normal phenotype. In X-linked recessive diseases, hemizygous males with a mutant allele will be affected, while the carrier female will not. Gene Therapy (GT) for recessive diseases has typically used a DNA-based gene system to deliver a normal allele in a mammalian expression vector or via allogenic cell transplant. However, numerous problems have been identified with these gene complementing systems arising from poor gene expression control, poor distribution of transfecting vector, and host immune responses to the viral vectors or cells. These adverse events as well as our own long-standing interest in DNA mismatch repair (MR), have led us to develop a new approach to gene therapy. In this alternative therapy, the existing mutant allele is targeted with a specifically designed synthetic oligonucleotide, which pairs with and induces correction of the chromosomal target via (MR). This alternative gene therapy can be called gene editing. In preliminary studies, we have begun to target the Sickle allele in a-globin found in CD34+ hematopoietic stem cells from patients with this recessive disease Sickle Cell Anemia (SCA) to produce a population of hematopoietic cells from diseased patients, which contain a normal 6-globin allele. This translational research is aimed at an IND for treatment of SCA patients via stem cell transplantation with their own edited cells. In this proposal we will apply this new technology to address inherited mutations in solid tissues. We will focus on X-linked recessive dystrophinopathies in muscle of animal models Duchenne dystrophy (DMD). This disease affects 1:3300 live born human males with severe, progressive muscle wasting, and is fatal by age 20-24 years of age. All attempts at complementing GT via viral and/or allogenic cell transplantation of either muscle or bone marrow precursor cells have provided no significant clinical improvement in patients. However, we have published a pilot study that demonstrated the feasibility of our gene editing in a canine model of DMD and the effects were long-term (11 months) albeit, subclinical. We therefore propose to follow up on these studies using the latest advances developed for the Sickle cell therapy paradigm. All studies in this Phase 1 proposal will relay on the ex vivo MR selection developed in yeast for SCA.

描述(申请人提供)：遗传性疾病，发生在身体的实体组织，通常是由简单的隐性突变引起的，源于两个表型正常的携带者父母。在X连锁隐性疾病中，带有突变等位基因的半合子男性会受到影响，而携带者女性不会受到影响。隐性疾病的基因治疗(GT)通常使用基于DNA的基因系统在哺乳动物表达载体中或通过同种异体细胞移植来传递正常等位基因。然而，由于基因表达控制不佳、载体分布不均匀以及宿主对病毒载体或细胞的免疫反应，这些基因互补系统存在许多问题。这些不良事件以及我们自己对DNA错配修复(MR)的长期兴趣，促使我们开发了一种新的基因治疗方法。在这种替代疗法中，现有的突变等位基因以专门设计的合成寡核苷酸为靶点，它与染色体靶标配对并通过(MR)诱导染色体靶标的纠正。这种另类基因疗法可以被称为基因编辑。在初步研究中，我们已经开始针对这种隐性疾病镰状细胞性贫血(SCA)患者CD34+造血干细胞中发现的a-珠蛋白中的镰刀等位基因，以产生一组来自疾病患者的造血细胞，其中包含正常的6-珠蛋白等位基因。这项翻译研究旨在通过干细胞移植治疗SCA患者的IND。在这项提案中，我们将应用这项新技术来解决固体组织中的遗传突变问题。我们将重点介绍Duchenne营养不良(DMD)动物模型肌肉中的X连锁隐性营养不良。这种疾病影响1：3300活体出生的男性，患有严重的进行性肌肉萎缩，在20-24岁时是致命的。所有通过肌肉或骨髓前体细胞的病毒和/或同种异体细胞移植来补充GT的尝试都没有在患者中提供显著的临床改善。然而，我们已经发表了一项初步研究，证明了我们在DMD犬模型中进行基因编辑的可行性，其效果是长期的(11个月)，尽管是亚临床的。因此，我们建议使用为镰刀细胞治疗范例开发的最新进展来跟踪这些研究。这一阶段1提案中的所有研究将依赖于在酵母中开发的用于SCA的体外MR选择。