TOPIC #366 - PHASE II SBIR CONTRACT - CLONOGENIC HIGH - THROUGHPUT ASSAY FOR SCREENING RADIATION MODULATION

克隆形成测定测量单细胞增殖和形成集落的能力。这一过程非常接近于放疗或化疗后肿瘤的再生长和复发，为筛选阻断这一过程的药物提供了一种检测方法。然而，克隆形成测定是劳动密集型的，并且对于化合物文库的高通量筛选（HTS）来说太麻烦。在这里，我们建议开发一个HTS系统的基础上克隆的端点。该提案基于已完成的第一阶段合同，其中我们成功地产生了适合小型化成多孔格式的头颈癌细胞，并进行了自动菌落计数。使用已知辐射调节剂的概念验证测试证明，该系统忠实地代表传统的克隆形成测定。在第二阶段的合同中，我们将通过化合物库进行筛选，产生额外的细胞系来代表其他癌症类型，并开发一个完全集成的自动化系统，该系统将细胞置于平板上，照射，暴露于药物，并计数菌落。该系统可移植到其他实验室。潜在的商业应用包括筛选具有抑制治疗后肿瘤再生长潜力的新化合物的文库，以及快速鉴定现有肿瘤药物中有效的成对组合。

Clonogenic assays measure the ability of single cells to proliferate and form a colony. This process approximates closely the regrowth and recurrence of tumors after treatment with radiation or chemotherapy, providing an assay to screen for drugs that block this process. Yet, clonogenic assays are labor-intensive and too cumbersome for high throughout screening (HTS) of compound libraries. Here we propose to develop an HTS system based on a clonogenic endpoint. This proposal is based on a completed Phase I contract wherein we successfully generated Head and Neck Cancer cells amenable to miniaturization into multi-well format with automated colony counting. Proof of concept tests using known radiation modulators demonstrate that this system faithfully represents the traditional clonogenic assays. In the Phase II contract, we will screen through compound libraries, generate additional cell lines to represent other cancer types, and develop a fully integrated automated system that plates cells, irradiates, exposes them to drug, and counts colonies. This system will be transferrable to other laboratories. Potential commercial applications include screening libraries for new compounds with the potential to inhibit regrowth of tumors after treatment and to rapidly identify efficacious pair-wise combinations among existing oncology agents.