Development of iPS Cells for Treatment of Hemoglobinopathies

开发用于治疗血红蛋白病的 iPS 细胞

• 批准号: 8710192
• 负责人: YUET Wai KAN
• 金额: $ 129.97万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-30 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8710192
• 关键词: Address; Adenoviruses; Advisory Committees; Africa; B-Lymphocytes; California; Cell Line; Cell Therapy; Cell Transplantation; Cells; Cellular biology; Collaborations; Complement; Complementary DNA; Data; Dedications; Development; Disease; Double-Stranded RNA; Economic Burden; Elements; Excision; Far East; Generations; Genes; Goals; Grant; Healthcare Systems; Hematopoietic; Hematopoietic System; Hematopoietic stem cells; Hemoglobinopathies; Hereditary Disease; Individual; Inherited; Integrase; Link; Mediating; Medical center; Methods; Middle East; Modification; Molecular Biology; Mutation; Oligonucleotides; Patients; Peptides; Plasmids; Polynucleotides; Quality of life; RNA; Regenerative Medicine; Research Institute; Resources; Safety; Science; Services; Sickle Cell; Sickle Cell Anemia; Somatic Cell; Stem Cell Research; Stem cell transplant; Stem cells; Structure of thyroid parafollicular cell; System; Testing; Thalassemia; Time; Xeno; Zinc Fingers; base; beta Globin; beta Thalassemia; c-Myc Staining Method; disease-causing mutation; gene therapy; homologous recombination; improved; induced pluripotent stem cell; nuclease; programs; reconstitution; social

DESCRIPTION (provided by applicant): The proposed POI is a multi-institutional grant that will develop a stem cell based therapy for the treatment of sickle cell disease (SCD) and ¿-thalassemia (¿-thal) as well as other hemoglobinopathies, using patient derived somatic cells and reprogramming them into induced pluripotent stem (IPS) cells that will have their mutations corrected and ultimately differentiated into hematopoietic stem cells (HSCs) to reconstitute the patient's hematopoietic system. Development of an effective cellular therapy for the treatment of hemoglobinopathies, the most common inherited diseases worldwide, would significantly improve the quality of life of individuals afflicted with SCD and B-thalassemia that are common among the peoples of Africa, the Mediterranean, the Middle East, and Asia as well as their descendents in the U.S. This proposal will test the hypothesis that an effective cellular and genetic therapy for these diseases can be achieved in the context of this PPG through the generation, modification, and the hematopoietic differentiation of patient derived iPS cells. This will be accomplished through the following Projects: Project 1 will involve the conversion of a patient's somatic cells into IPS cells using phiC31 Integrase-mediated, sequence-specific integration of a plasmid carrying 2A peptide linked Oct4, Sox2, Klf4, and cMyc reprogramming cDNAs or by using small activating double stranded RNA (saRNA) to transiently enhance the expression of these reprogramming genes. Project 2 will involve correction of the disease causing mutations in the somatic cells and the iPS cells by sequence specific modification using either classical homologous recombination (HR) or by oligo/polynucleotide-based small fragment homologous replacement (SFHR) in the presence or absence of targeted zinc finger nucleases (ZFNs) or other meganucleases. Project 3 will involve exposure of uncorrected and corrected iPS cells to conditions to direct hematopoietic differentiation to generate HSCs which have the capacity to engraft and reconstitute the hematopoietic system. In the course of this PPG, all Projects will develop xeno-free systems to optimize safety. The science in the Projects will be augmented by an administrative (Core A) and 2 scientific Cores: Core B: Cell and Molecular Biology, and Core C: Cell Transplantation and Analysis.

描述（由申请人提供）：拟议的 POI 是一项多机构拨款，将开发一种基于干细胞的疗法，用于治疗镰状细胞病 (SCD) 和 β 地中海贫血 (¿-thal) 以及其他血红蛋白病，使用患者来源的体细胞并将其重新编程为诱导多能干 (IPS) 细胞，这些细胞将纠正其突变并最终分化为造血细胞 干细胞（HSC）来重建患者的造血系统。开发一种有效的细胞疗法来治疗血红蛋白病（全世界最常见的遗传性疾病），将显着改善患有 SCD 和 B 地中海贫血的患者的生活质量，这些疾病在非洲、地中海、中东和亚洲及其在美国的后代中很常见。这项提案将检验这样的假设：针对这些疾病的有效细胞和基因疗法可以 在此 PPG 的背景下，通过患者来源的 iPS 细胞的生成、修饰和造血分化来实现。这将通过以下项目来完成：项目 1 将涉及使用 phiC31 整合酶介导的序列特异性整合携带 2A 肽连接 Oct4、Sox2、Klf4 和 cMyc 重编程 cDNA 的质粒，或通过使用小激活双链 RNA (saRNA) 暂时增强这些重编程基因的表达，将患者的体细胞转化为 IPS 细胞。项目 2 将涉及在存在或不存在靶向锌指核酸酶 (ZFN) 或其他大范围核酸酶的情况下，通过使用经典同源重组 (HR) 或基于寡核苷酸/多核苷酸的小片段同源替换 (SFHR) 的序列特异性修饰来纠正引起体细胞和 iPS 细胞突变的疾病。项目3将涉及将未校正和校正的iPS细胞暴露于指导造血分化的条件下，以产生具有移植和重建造血系统能力的HSC。在此 PPG 过程中，所有项目都将开发无异种物质系统以优化安全性。项目中的科学将通过行政（核心 A）和 2 个科学核心得到增强：核心 B：细胞和分子生物学，以及核心 C：细胞移植和分析。