Discovery of Death Ligands Against Cancers

描述（由申请人提供）：最近，我们通过将已知的细胞粘附（或细胞捕获）配体添加到OBOC文库中的每个珠的表面来修改一珠一化合物（OBOC）组合文库方法。当将活细胞与这种新的一珠两化合物（OB2C）文库一起孵育时，面向珠表面的捕获细胞的细胞膜暴露于每个珠上展示的文库化合物。使用适当的报告系统，应该能够快速检测可以引发特定生化或细胞反应的珠粒（激动剂）。类似地，如果细胞被外源激动剂刺激，则也可以用这种方法发现抑制特定生物化学或细胞反应的分子（拮抗剂）。在这个R33提案中，我们计划将我们的努力集中在使用新的OB2C技术来发现针对血液和实体恶性肿瘤的促凋亡细胞表面作用分子。本项目的具体目标如下：目标1：设计和合成OB2C组合文库，用于发现合成的和细胞表面作用的促凋亡分子或死亡配体。目标二：筛选OB2C组合文库，以发现针对淋巴癌、急性髓性白血病和实体瘤细胞的合成的和细胞表面作用的死亡配体。先导化合物将进一步优化与重点OB2C组合库。目标3：重新合成先导化合物并评价其自身或与癌细胞表面靶向配体缀合后的促凋亡功能。将确定这些促凋亡剂的作用机制。影响：超高通量OB2C文库方法是高效和经济的。一旦优化，它可以很容易地被许多学术研究人员应用到他们的研究。在这项拟议的研究中开发的死亡配体可以开发成新的有效但毒性较小的癌症治疗剂。这些配体也可用作基础癌症生物学和系统生物学研究的生物活性探针，特别是在它们的作用机制已经阐明之后。

DESCRIPTION (provided by applicant): Recently, we have modified the one-bead-one-compound (OBOC) combinatorial library method by adding a known cell adhesion (or cell capturing) ligand to the surface of every bead in the OBOC library. When live cells are incubated with such novel one-bead-two-compound (OB2C) libraries, the cell membranes of the captured cells facing the bead surface are exposed to the library compounds displayed on each bead. With an appropriate reporter system, one should be able to rapidly detect beads that can elicit a specific biochemical or cellular response (agonists). Similarly, if the cells are stimulatd by an exogenous agonist, molecules that suppress specific biochemical or cellular response (antagonists) can also be discovered with this approach. In this R33 proposal, we plan to focus our effort on using the novel OB2C technology to discover pro- apoptotic cell surface acting molecules against both hematologic and solid malignancies. Specific aims of this proposed project are as follows: Aim 1: To design and synthesize OB2C combinatorial libraries for the discovery of synthetic and cell surface acting pro-apoptotic molecules or death ligands. Aim 2: To screen OB2C combinatorial libraries for the discovery of synthetic and cell surface acting death ligands against lymphoid cancer, acute myeloid leukemia and solid tumor cells. The lead compounds will be further optimized with focused OB2C combinatorial libraries. Aim 3: To resynthesize the lead compounds and evaluate their pro-apoptotic functions by themselves or after conjugation to the cancer cell surface targeting ligands. The mechanisms of action of these pro-apoptotic agents will be determined. Impact: The ultra-high throughput OB2C library method is highly efficient and economical. Once optimized, it can be readily applied by many academic investigators to their research. The death ligands to be developed in this proposed research can be developed into novel effective but less toxic cancer therapeutics. These ligands may also be used as biologically active probes for basic cancer biology and systems biology research, particularly after their mechanisms of action have been elucidated.