SELECTIVE APPROACHES TO HEMATOPOIETIC STEM CELL AGONISTS

成人造血起源于原始多能干细胞， 骨髓造血干细胞的体外扩增是一种有效的方法。 实验和临床血液学的主要目标。事实上，基因治疗 原始造血细胞的能力受到阻碍， 常规地在培养物中扩增干细胞。本提案的目的 是开发一个新的和新颖的屏幕，将确定因素， 导致骨髓干细胞自我更新。在第一阶段，我们将建造 组合文库，并鉴定特异性结合到细胞表面的配体。 原始人和小鼠造血干细胞和小鼠干细胞 线在此之后，我们将设计和执行高通量筛选 鉴定在增殖中作为激动剂的模拟分子， 造血干细胞系。在第二阶段，个别候选人将 在各种测定中对原代人原始造血细胞进行测试， 细胞 这些分析应该区分真正的干细胞更新， 增殖与血统承诺相结合。 拟议的商业应用： 在第一阶段确定的激动剂将提供给医疗和 基因治疗试剂的形式研究社区扩大 干/祖细胞在体外，以改善造血转导 干/祖细胞通过逆转录病毒载体用于持续治疗 好处，以提高血细胞生产在体内的成本效益 用于骨髓抑制和剂量强化治疗的药物 作为免疫诊断试剂，细胞分离， 以及造血的基本机制研究。图书馆 也应该是一个丰富的，新的，廉价的来源标记干， 祖细胞分离。

Adult hematopoiesis originates from primitive pluripotent stem cells in the bone marrow. In vitro expansion of hematopoietic stem cells is a major goal of experimental and clinical hematology. In fact, gene therapy of primitive hematopoietic cells has been hindered by the inability to routinely expand stem cells in culture. The objective of this proposal is to develop a new and novel screen that will identify factors that cause bone marrow stem cells to self renew. In phase I, we will construct combinatorial libraries, and identify ligands specific to the surface of primitive human and murine hematopoietic stem cells and murine stem cell lines. Following this, we will design and perform high throughput screens to identify mimetic molecules that behave as agonists in proliferation of hematopoietic stem cell lines. In phase II, individual candidates will be tested in a variety of assays on primary human primitive hematopoietic cells. These assays should distinguish true stem cell renewal from proliferation in conjunction with lineage commitment. PROPOSED COMMERCIAL APPLICATIONS: Agonists identified in phase I will be provided to the medical and research community in the form of reagents for gene therapy to expand stem/progenitor cells in vitro, to improve transduction of hematopoietic stem/progenitor cells by retroviral vectors for sustained therapeutic benefit, to improve blood cell production in vivo for cost-effective therapeutics in myelosuppression and for dose-intensification treatment of various diseases, as reagents for immunodiagnostics, cell separation, and research on fundamental mechanisms in hematopoiesis. The libraries should also be a rich, new, inexpensive source of markers for stem and progenitor cell separation.