Gene Transfer into Hematopoietic Stem Cells

基因转移至造血干细胞

• 批准号: 6681510
• 负责人: Fabio Candotti
• 金额: --
• 依托单位: NATIONAL HUMAN GENOME RESEARCH INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6681510
• 关键词: AIDS; AIDS /HIV diagnosis; HIV infections; blood disorder; blood disorder chemotherapy; bone marrow; cell differentiation; clinical research; cord blood; gene therapy; genetic manipulation; genetic regulation; hematopoietic stem cells; hematopoietic tissue transplantation; human subject; laboratory mouse; leukemia; sheep; stem cell transplantation; thalassemia; tissue /cell culture; transplantation immunology

A variety of inherited and acquired human diseases affecting the hematopoietic stem cell (HSC) can potentially be treated with genetic manipulation of the patients? HSCs. Potential applications of this technology include therapeutic approaches for common blood disorders (e.g. thalassemia), malignancies (e.g. leukemia) and infectious diseases of the blood cells, such as HIV-1 infection/AIDS. However, with a few exceptions, gene transfer into HSCs of large animal models and humans has thus far proven to be difficult and inefficient. By improving our knowledge of the biological characteristics of the human HSC we will likely gain important insights that will allow us to improve the efficiency of gene transfer in and correction of this elusive target cell. Under this project we have established in vitro and in vivo animal models of human hematopoiesis using hematopoietic progenitors obtained from cord blood, bone marrow or peripheral blood of volunteer donors. These cells are cultured in vitro in a variety of cytokines and growth factors to identify conditions that allow their survival in the absence of differentiation. The cells are then subjected to gene transfer and allowed to differentiate into mature cell lineages in vitro (using colony forming unit or T cell differentiation assays) or in vivo, using mouse or sheep animal models. These experiments allow comparison of cells with putative hematopoietic stem cell activity isolated from various sources (e.g. cord blood vs. bone marrow vs. peripheral blood) and testing of vectors and gene transfer conditions that efficiently target these cells.

可能通过对患者的遗传操纵来治疗各种影响造血干细胞（HSC）的遗传和获得的人类疾病？ HSC。这项技术的潜在应用包括常见血液疾病（例如，thalassya），恶性肿瘤（例如白血病）和血细胞感染性疾病（例如HIV-1感染/艾滋病）的治疗方法。但是，除少数例外，迄今为止，基因转移到大型动物模型和人类的HSC中是困难和效率低下的。通过提高我们对人类HSC生物学特征的了解，我们可能会获得重要的见解，从而使我们能够提高基因转移的效率，并纠正该难以捉摸的靶细胞。在这个项目下，我们使用从绳索血液，骨髓或志愿供体的外周血中获得的造血祖细胞建立了人类造血的体外和体内动物模型。这些细胞在各种细胞因子和生长因子中在体外进行培养，以鉴定允许在没有分化的情况下存活的疾病。然后，将细胞经过基因转移，并在体外（使用菌落形成单元或T细胞分化测定法）或使用小鼠或绵羊动物模型分化为成熟的细胞谱系（使用菌落形成单元或T细胞分化测定）。这些实验允许将细胞与假定的造血干细胞活性进行比较，这些干细胞活性是从各种来源分离的（例如脐带血与骨髓与外周血），并测试矢量和基因转移条件，有效地靶向这些细胞。