Hematopoietic stem cell gene therapy for sickle cell disease

镰状细胞病的造血干细胞基因治疗

• 批准号: 7821229
• 负责人: DEREK A PERSONS
• 金额: $ 30.43万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7821229
• 关键词: Address; Age-Years; Animal Model; Biological Assay; Blood Circulation; Bone Marrow; Bone Marrow Cells; Bone Marrow Transplantation; Busulfan; Cell Culture Techniques; Cells; Cessation of life; Clinical; Clinical Trials; Clone Cells; Data; Development; Disease; Engraftment; Enrollment; Erythroid; Erythroid Cells; Erythropoiesis; Funding; Future; Gene Expression; Gene Targeting; Gene Transfer; Generations; Genes; Globin; Goals; Hematological Disease; Hematopoietic; Hematopoietic stem cells; Hemoglobin; Homologous Gene; Human; Immunodeficient Mouse; Individual; Inherited; Insertional Mutagenesis; Intervention; Learning; Lentivirus Vector; Macaca mulatta; Mediating; Methods; Mission; Modeling; Mouse Strains; Mus; Myelogenous; Myeloid Cells; National Heart, Lung, and Blood Institute; Patients; Peripheral; Phase; Pre-Clinical Model; Preparation; Process; Production; Public Health; Recombinants; Regulatory Element; Research; Retroviridae; Risk; Safety; Series; Sickle Cell; Sickle Cell Anemia; Stem cells; Symptoms; Syndrome; Testing; Thalassemia; Thalassemia intermedia; Therapeutic; Toxic effect; Translational Research; Transplantation; Treatment Protocols; Viral; adeno-associated viral vector; base; beta Thalassemia; conditioning; cytokine; disability; env Gene Products; gene therapy; gene therapy clinical trial; genetically modified cells; granulocyte; immunodeficient mouse model; improved; lambda Spi-1; leukemia; particle; pre-clinical; programs; research study; success; therapeutic gene; tumor; vector; vector control; young adult

The goals of this project are: 1) to identify the optimal envelope protein for vector preparation and to develop a strategy for preparing and concentrating Y-globin lentiviral vector particles that allow efficient gene transfer into primitive human hematopoietic cells from patients with sickle cell disease and (3-thalassemia that engraft in immunodeficient mice; 2) to evaluate the safety of our Y-globin lentiviral vector in cell culture and animal models and 3) to initiate a Phase I/I I clinical gene therapy trial for severe (3-thalassemia and sickle cell disease. Our Specific Aims are as follows: 1: to obtain therapeutic levels of globin lentiviral vectormediated gene transfer into primitive human hematopoietic cells; 2: to evaluate the safety of globin lentiviral vectors using preclinical models; and 3: to evaluate stem cell-targeted, globin lentiviral vector-mediated gene transfer and expression in human patients with p-thalassemia intermedia, HbE-p-thalassemia, homozygous sickle cell anemia, or HbS-p-thalassemia. In pursuing the first aim, we will develop a producer cell clone for our Y-globin lentiviral vector that yields high titer vector preparations and will develop methods for purifying and concentrating vector particles. We will also identify the appropriate envelope protein pseudotype that, after the purification and concentration of vector particles, insures efficient gene transfer into primitive human hematopoietic cells as assayed in an immunodeficient mouse model. Because the risk of insertional mutagenesis is inherent to the use of integrating retroviruses and has resulted in the development of leukemia in gene therapy trials, we have planned a comprehensive series of experiments to evaluate the safety of Y-globin lentiviral vector-mediated gene transfer. Our preliminary data suggests that globin vectors are safer than vectors used in prior clinical trials because erythroid-specific, rather than powerful viral regulatory elements that are active in stem cells, are used to drive therapeutic gene expression. We propose to continue testing this hypothesis. We anticipate that the needed preclinical data from these first 2 specific aims will be obtained during the first 2-3 years of funding and will support a proposed phase I/I I clinical gene therapy trial beginning in year 3, initially for patients with severe (3-thalassemia who have a-globin excess, and then if success is achieved, for young adult patients with severe sickle cell disease. While our goal is that a clinical benefit will be obtained for individual patients, we are certain to learn important scientific information that will advance the likelihood of clinical success in future trials for the hemoglobin disorders. Our goals are relevant to public health and the mission of the National Heart, Lung and Blood Institute since the development of efficient stem cell targeted gene transfer would provide therapy for many inherited blood diseases. Because sickle cell disease causes severe symptoms, disability and often early death, curative therapies such as gene therapy are urgently needed.

本课题的主要目标是：1）筛选出最适合于载体制备的包膜蛋白， 制备和浓缩允许有效基因转移的Y-珠蛋白慢病毒载体颗粒的策略 从患有镰状细胞病和β-地中海贫血的患者体内植入原始的人类造血细胞， 在免疫缺陷小鼠中; 2）评估我们的Y-珠蛋白慢病毒载体在细胞培养和动物中的安全性 模型和3）启动I/II期临床基因治疗试验，用于严重（β-地中海贫血和镰状细胞贫血） 疾病我们的具体目标如下：1.获得治疗水平的珠蛋白慢病毒载体介导的 基因转移入原始人造血细胞; 2：评价珠蛋白的安全性 慢病毒载体，使用临床前模型;和3：评估干细胞靶向的球蛋白慢病毒 在中间型β地中海贫血患者中载体介导的基因转移和表达， HbE-p-地中海贫血、纯合子镰状细胞贫血或HbS-p-地中海贫血。为了实现第一个目标， 我们将开发Y-球蛋白慢病毒载体的生产细胞克隆，产生高滴度载体制剂 并将开发提纯和浓缩载体颗粒的方法。我们还将确定 合适的包膜蛋白假型，在载体颗粒纯化和浓缩后， 确保有效的基因转移到原始人造血细胞中，如在免疫缺陷的 小鼠模型因为插入突变的风险是使用整合逆转录病毒所固有的 并导致了白血病基因治疗试验的发展，我们已经计划了一个全面的 一系列实验来评估Y-珠蛋白慢病毒载体介导的基因转移的安全性。我们 初步数据表明，珠蛋白载体比先前临床试验中使用的载体更安全， 红系特异性的，而不是在干细胞中有活性的强大的病毒调节元件，被用于 驱动治疗基因的表达。我们建议继续检验这一假设。我们预计 前2个特定目标所需的临床前数据将在资助的前2-3年内获得 并将支持拟议的I/III期临床基因治疗试验，最初用于患有以下疾病的患者： 严重（3-地中海贫血谁有a-珠蛋白过量，然后如果成功，对于年轻的成年患者 患有严重的镰状细胞病虽然我们的目标是为个体患者获得临床益处， 我们肯定会学到重要的科学信息，这些信息将提高临床成功的可能性， 血红蛋白紊乱的未来试验我们的目标与公共卫生和卫生组织的使命有关。 国家心肺血液研究所自开发高效干细胞靶向基因转移 将为许多遗传性血液病提供治疗。因为镰状细胞病会导致严重的 在出现症状、残疾和往往过早死亡的情况下，迫切需要诸如基因疗法等治愈性疗法。