Novel High Content Vascular Patterning Assay(RMI)

血管生成，即新血管的生长，发生在各种人类疾病中。即使 抗血管生成药物仍然是一个巨大的市场，以高含量高通量细胞为基础的 对于这一应用，发现血管生成抑制物的检测方法仍有待开发。我们相信 开发新的检测方法作为筛选工具，以发现靶向内皮细胞分化的抑制剂 可以被证明是识别新类别抗血管生成药物的有效手段。在此，我们建议 建立一种可用于高通量筛选的高含量血管花纹分析方法 (HTS)。我们计划通过接口High来构建一个功能强大的高内容筛选平台的原型 内皮细胞构型的多参数光学成像的吞吐量筛查能力 维(3-D)矩阵。我们实现这些目标的创新方法将通过以下方式实现 目标有以下几点。在特定目标1中，我们将开发并验证一种三维内皮细胞图案化试验(3- D ECPA)。我们将通过开发产生高度健壮的多细胞内皮细胞的方法来实现这一点 在微载体珠上的细胞球体，并在384孔光学中将球体排列在3-D胶原基质中 放映板。此外，这些检测将在低氧环境下进行。《特定目标2》 我们计划开发和验证一个高内容(HC)图像分析平台，用于3D ECPA。这 将通过开发软件来识别3D中内皮细胞图案的新特征来实现 并应用自动分析来获得形态发现指纹。最后，我们将验证3D 用于高温超导应用的ECPA和图像分析软件，通过筛选一套多样化的天然产品。我们 相信这个HC-HTS平台将提供筛选大量小分子独特机会 血管生成的抑制剂，并协助药物发现。

Angiogenesis, the growth of new blood vessels, occurs in a diverse array of human diseases. Even though anti-angiogenesis remains a large market for Pharmaceuticals, high content high throughput cell based assays for angiogenesis inhibitor discovery remain to be developed for this application. It is our belief that developing new assays as screening tools for discovery of inhibitors that target endothelial cell differentiation can prove to be an efficient means of identifying new classes of anti-angiogenic drugs. Here we propose to develop a high content vascular patterning assay that can likely be adopted for high throughput screening (HTS). We plan to build a prototype of a powerful high content screening platform by interfacing high throughput screening capability with multiparameter optical imaging of endothelial cell patterning in three- dimensional (3-D)matrices. Our innovative approach to achieve these goals will be accomplished through the following aims. In Specific aim 1 we will develop and validate a 3-D endothelial cell patterning assay (3- D ECPA). We will accomplish this by developing methods to generate highly robust multicellular endothelial cell spheroids on microcarrier beads and array the spheroids in 3-D collagen matrix in 384 well optical screening plates. In addition, these assays will be performed under hypoxic environment. In Specific Aim 2 we plan to develop and validate a high content (HC)image analysis platform for use with 3-D ECPA . This will be accomplished by developing software that identifies novel features of endothelial cell patterning in 3-D matrix and apply automated analysis to obtain morphogenic findgerprints. Finally, we will validate the 3-D ECPA and image analysis software for HTS application by screening a diversity set of natural products. We beleve that this HC-HTS platform will offer unique opportunities to screen vast numbers of small molecule inhibitors of angiogenesis and assist in drug discovery.