Human embryonic stem (hES) cells for hematopoietic and HIV/AIDS therapies

描述（由申请人提供）：用于造血和HIV/AIDS治疗的人胚胎干细胞（hESC）由于其能够在体内产生任何类型的细胞以及易于进行基因修饰的能力，在新型细胞替代疗法中显示出巨大的希望。目前，从不可再生来源如骨髓和脐带血中获得的CD34造血祖细胞（HPC）被用于临床应用。最近的研究表明，CD34造血祖细胞（hES-CD34）可以从hESC中提取，这为其在造血细胞和艾滋病治疗中的应用开辟了许多令人兴奋的可能性。作为实现这一目标的第一步，我们最近成功地从hES-CD34细胞中获得了功能正常的巨噬细胞和树突状细胞，并表明它们对HIV-1感染敏感，从而为在该系统中测试各种抗hiv基因治疗构建铺平了道路。由于缺乏理想的动物模型，对hES-CD34细胞在体内的多系造血能力进行全面评估是不可能的。最近出现了一种新的人源化小鼠模型（ragu -hu），允许这样的评估，我们已经在我们的实验室建立了这个系统。在我们正在进行的基因治疗研究中，我们也发现了一些新一代的高效抗hiv基因结构。这些新进展为评价hES-CD34细胞在基因修饰、多系造血和HIV基因治疗策略中的应用奠定了良好的基础。在本提案中，我们的目标是利用新兴技术，并在我们最近的进展基础上，将hESC衍生细胞转化为临床应用。我们建议的具体目标是：1。用抗hiv基因对hESC进行遗传编程，获得抗HIV-1的成熟造血终末期细胞。2. 评价Rag2-/-3c-/-小鼠hES-CD34细胞的多系造血分化能力。

DESCRIPTION (provided by applicant): Human embryonic stem cells (hESC) for hematopoietic and HIV/AIDS therapies Human embryonic stem cells (hESC) show great promise for novel cell replacement therapies due to their capacity to give rise to any type of cell in the body and their tractability for genetic modification. Currently, CD34 hematopoietic progenitor cells (HPC) obtained from non-renewable sources such as bone marrow and cord blood are used for clinical applications. The recent demonstration that CD34 hematopoietic progenitor cells (hES-CD34) could be derived from hESC opened up many exciting possibilities for their use in hematopoietic cell and AIDS therapies. As a first step towards this goal, we succeeded in our recent efforts in deriving functionally normal macrophages and dendritic cells from hES-CD34 cells and showed that they are susceptible to HIV-1 infection thus paving the way for testing various anti-HIV gene therapeutic constructs in this system. Due to the lack of an ideal animal model, a thorough assessment of the multi-lineage hematopoietic capacity of hES-CD34 cells in vivo has not been possible. A novel humanized mouse model (RAG-hu) has emerged recently that permits such an evaluation, and we have this system currently established in our laboratory. We also identified a number of new generation anti-HIV gene constructs with high efficacy in our ongoing gene therapy studies. These new developments laid a good foundation to evaluate the utility of hES-CD34 cells for gene modification, multi-lineage hematopoiesis and HIV gene therapy strategies. In this proposal our aim is to exploit the newly emerging technologies and build upon our recent progress towards translating hESC derived cells for clinical applications. The specific objectives of our proposal are to: 1. Genetically program hESC with anti-HIV genes and derive HIV-1 resistant mature hematopoietic end stage cells. 2. Evaluate the multi-lineage hematopoietic differentiation capacity of hES-CD34 cells in Rag2-/-3c-/- mice. PUBLIC HEALTH RELEVANCE: Human embryonic stem cells show enormous promise for developing novel cell replacement and gene therapies. In these proposed studies, anti-HIV genes will be introduced into stem cells to derive virus resistant white blood cells. Success of this work would not only benefit HIV/AIDS field but also will have broader implications for hematopoietic cell therapies and transplantation.