TARGETED MANIPULATION OF STEM CELLS FOR AIDS THERAPY

描述：造血干细胞是一种有效的治疗工具，能够 完全重建移植宿主的免疫功能。遗传 干细胞的操作代表了一种产生免疫功能的方法 对艾滋病毒感染的抵抗力，从而有可能恢复免疫 仅靠HAART就可以超越这一可能。限制了实际应用 干细胞为基础的治疗是植入的低效率施加 对分离和转导方案的不切实际的量化要求。这个 体外扩增干细胞的能力将克服 领域，但目前的协议涉及细胞因子刺激，通常 牺牲多潜能来换取更多的细胞数量。此应用程序 提出一种显性抑制调控干细胞增殖的模型 使用经过改造的小鼠发出信号并提供数据以支持模型 缺乏细胞周期蛋白依赖性激酶抑制因子(CDKI)p21。的目标是 本申请的目的是： 1)。进一步确定干细胞中细胞周期抑制的近似性介体 重点放在a)p21家族成员p27，使用基因靶向小鼠和b) 抑制性转化生长因子-b信号通路与p21、p27的相互关系。 2)。改变细胞周期抑制控制作为诱导细胞的一种手段 增殖，测试细胞是否通过这种机制进入细胞周期 避免细胞因子的促分化作用。解除链接的可能性 通过增殖分化实现干细胞的体外扩增 将会被定义。 3)。确定HIV-1暴露是否改变细胞周期抑制物的表达 或影响静止的干细胞对CDKI的应答 下调制。通过靶向细胞操纵干细胞的潜力 在特定情况下作为干细胞扩增手段的周期抑制物 艾滋病毒疾病将被定义。 这些目标的实现将指导干细胞基因的具体策略 基于干细胞调节原理的艾滋病毒疾病的治疗。

Description: Hematopoietic stem cells are a potent therapeutic tool capable of fully reconstituting immunologic function in a transplanted host. Genetic manipulation of stem cells represents a means of generating immune function resistant to HIV infection and thereby the potential of immune restitution beyond that possible with HAART alone. Limiting the practical application of stem cell based therapeutics is the low efficiency of engraftment imposing unrealistic quantitative demands on isolation and transduction protocols. The ability to expand stem cells ex vivo would overcome major limitations of the field, but current protocols involve cytokine stimulation that generally sacrifice multipotentiality for increased cell numbers. This application proposes a model of stem cell proliferation regulated by dominant inhibitory signals and presents data to support the model using mice engineered to be deficient in the cyclin dependent kinase inhibitor (CDKI), p21. The aims of this application are to: 1). Further define proximate mediators of cell cycle inhibition in stem cells focusing on a) the p21 family member, p27, using gene targeted mice and b) the interrelationship of the inhibitory TGF-b signaling pathway with p21 and p27. 2). Alter cell cycle inhibitory control as a means of inducing cell proliferation, testing whether cells entering cell cycle by this mechanism avoid the pro-differentiative effect of cytokines. The potential for unlinking proliferation and differentiation to achieve ex vivo expansion of stem cells will be defined. 3). Determine if HIV-1 exposure alters the expression of cell cycle inhibitors in stem cells or affects quiescent stem cell responsiveness to CDKI down-modulation. The potential of manipulating stem cells by targeting cell cycle inhibitors as a means of expanding stem cells in the specific context of HIV disease will be defined. Accomplishment of these aims will guide specific strategies for stem cell gene therapy of HIV disease based on principles of stem cell regulation.