Derivation and Correction of Thalassemic Pluripotent Stem Cells

地中海贫血多能干细胞的衍生和校正

• 批准号: 7799411
• 负责人: David W Russell
• 金额: $ 45.09万
• 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: 2009-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7799411
• 关键词: Adult; Autologous Transplantation; Biological Assay; CD34 gene; Cell Line; Cells; Clinical; Coculture Techniques; DNA Methylation; Deoxyribonucleases; Dependovirus; Derivation procedure; Development; Disease; Engraftment; Epigenetic Process; Erythroid; Erythroid Cells; Excision; Fibroblasts; Funding; Gene Delivery; Gene Targeting; Genes; Genetic Recombination; Globin; Grant; Hematopoietic; Hematopoietic stem cells; Hemoglobin F Disease; Hereditary Disease; Human; Hypersensitivity; Immunodeficient Mouse; In Vitro; Integrase; Kanamycin Kinase; Lentivirus Vector; Long Terminal Repeats; Mediating; Mesenchymal; Methods; Monitor; Mus; Mutation; Patients; Phenotype; Pluripotent Stem Cells; Point Mutation; Production; Proteins; Recovery; Research; Retroviral Vector; Site; Source; Spumavirus; Stem cells; Stromal Cells; Subfamily lentivirinae; Surface; System; Testing; Thalassemia; Therapeutic; Thymidine Kinase; Transgenes; Transplantation; adeno-associated viral vector; arm; base; beta Globin; blastomere structure; cell type; design; disease-causing mutation; erythroid differentiation; fetal; fusion gene; gene correction; gene therapy; genome-wide; genotoxicity; histone modification; homologous recombination; human embryonic stem cell; improved; induced pluripotent stem cell; mutant; pluripotency; recombinase; research study; small hairpin RNA; vector

Human induced pluripotent stem cells (iPSCs) have the potential to treat many diseases by autologous transplantation. However, before they can be used clinically, efficient and reproducible methods are required for their derivation and differentiation into therapeutic cell types. In the case of genetic diseases, methods for correcting disease-causing mutations also need to be developed. Here we will use this approach to design a treatment for thalassemia, which is caused by mutations in globin genes. This approach avoids the potential genotoxic complications of conventional gene therapy with retroviral vectors and should achieve consistent, regulated globin expression from the endogenous locus. We will derive IPSCs from the adult cells of patients with thalassemia using lentivirus and foamy virus vectors that express reprogramming transgenes. Some of the vectors will be designed for transient gene delivery to create transgene-free iPSCs. These IPSCs will be differentiated into hematopoietic cells to determine the best adult cell type and reprogramming vectors for creating iPSCs with hematopoietic potential. Adeno-associated virus gene targeting vectors will be used to correct the globin mutations in these thalassemic IPSCs, and globin expression will be studied after their differentiation into erythroid progeny. This research plan capitalizes on recent advances in the derivation of patient-specific stem cells, which in combination with gene correction constitutes a new paradigm for the treatment of genetic diseases.

人诱导的多能干细胞（IPSC）具有自体治疗许多疾病的潜力 移植。但是，在需要临床，高效和可重复的方法之前 因为它们的推导和分化为治疗细胞类型。在遗传疾病的情况下， 还需要开发纠正致病突变。在这里，我们将使用这种方法设计 地中海贫血的治疗是由球蛋白基因突变引起的。这种方法避免了潜力 常规基因治疗与逆转录病毒载体的遗传毒性并发症，应达到一致的， 来自内源基因座的球蛋白表达。我们将从患者的成年细胞中得出IPSC 使用慢病毒和泡沫病毒载体，表达重编程转基因。其中一些 向量将设计用于瞬态基因递送，以创建不含转基因的IPSC。这些IPSC将是 分化为造血细胞，以确定最佳的成年细胞类型和重编程向量 创造具有造血潜力的IPSC。腺相关的病毒基因靶向载体将用于 纠正这些丘脑血症IPSC中的球蛋白突变，并在其之后研究球蛋白的表达 分化为红细胞后代。该研究计划利用了最新的推导进步 患者特异性干细胞与基因校正结合构成了新的范式 遗传疾病的治疗。