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

描述(申请人提供)：人类胚胎干细胞(HESC)用于造血和艾滋病毒/艾滋病治疗人类胚胎干细胞(HESC)因其能够在体内产生任何类型的细胞以及易于处理进行基因修改而在新的细胞替代疗法中显示出巨大的前景。目前，从骨髓、脐带血等不可再生来源获得的CD34造血祖细胞(HPC)正在用于临床应用。最近发现的CD34造血祖细胞(HES-CD34)可以从hESC分化而来，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。作为迈向这一目标的第一步，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基因治疗结构铺平了道路。由于缺乏理想的动物模型，在体内对HES-CD34细胞的多系造血能力的彻底评估是不可能的。最近出现了一种新的人源化小鼠模型(RAG-HU)，它允许进行这样的评估，我们目前在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，我们还确定了一些新一代高效的抗HIV基因构建物。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。在这项建议中，我们的目标是利用新出现的技术，并在我们最近在将hESC来源的细胞转化为临床应用方面取得进展的基础上。我们建议的具体目标是：1.用抗HIV基因对hESC进行基因编程，获得抗HIV-1的成熟造血终末期细胞。2.评价HES-CD34细胞在Rag2-/-3c-/-小鼠体内的多向分化能力。 与公共卫生相关：人类胚胎干细胞在开发新的细胞替代和基因疗法方面显示出巨大的前景。在这些拟议的研究中，将把抗艾滋病毒基因引入干细胞，以获得抗病毒的白细胞。这项工作的成功不仅将使艾滋病毒/艾滋病领域受益，而且将对造血细胞治疗和移植产生更广泛的影响。

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.