Orthotopic models of tumor angiogenesis and blood flow

描述（由申请人提供）：需要肿瘤血流、血管内癌细胞运输和外渗的临床相关可成像动物模型，这些模型是当前药物评价和开发的关键目标。肿瘤细胞运输和血流的大多数模型或者基于肿瘤的皮下移植或者基于背侧皮褶窗模型。这些模型通常不允许转移。也有证据表明，肿瘤血管分布可能是器官部位特异性的，因此原位模型将是非常重要的。我们先前已经开发了表达绿色荧光蛋白（GFP）的癌症的原位转移模型（Nature Rev Cancer 5，796-806，2005）。我们已经表明，这些模型能够对肿瘤生长和转移以及内脏器官上的血管生成进行高分辨率全身成像（PNAS 97，1206-11，2000; 99，3824-9，2002）。I期的新进展包括构建具有在细胞核中的GFP和在细胞质中的RFP的双色癌细胞（Cancer Res 64，4251-6，2004; 65，4246-52，2005），其能够在活小鼠中成像肿瘤细胞迁移、核-细胞质动力学和外渗。具有表达GFP的血管的裸鼠模型也已经在I期开发，其可以促进肿瘤血流的成像（Cancer Res 64，8651-6，2004; 65，5352-7，2005）。随着这些技术在第一阶段赠款期间开发，第二阶段赠款将进一步开发原位模型中的这些技术，以评估肿瘤血流，血管内肿瘤细胞运输和外渗的抑制剂。具体目标包括：（2）使用原位植入裸鼠中的在细胞核中具有GFP且在细胞质中具有RFP的双色癌细胞用于体内测试靶向肿瘤细胞的血管内运输的试剂;（3）原位植入裸鼠中的在细胞核中具有GFP且在细胞质中具有RFP的双色癌细胞用于体内测试靶向外渗肿瘤细胞的试剂的用途。这些原位模型的独特之处在于，它们将能够在临床相关小鼠模型中在核质动态水平上可视化肿瘤血流、血管内肿瘤细胞运输和外渗。新开发的Olympus OV 100全鼠成像系统具有用于宏观和高分辨率亚细胞成像的光学器件，沿着模型，以筛选针对这些转移关键步骤的初始一组潜在抑制剂。这些模型将在第三阶段商业化使用，以筛选和评估大量的药物和化合物库。需要肿瘤血流、血管内癌细胞运输和外渗的临床相关的可成像动物模型，其是当前药物评价和开发的关键目标。肿瘤细胞运输和血流的大多数模型或者基于肿瘤的皮下移植或者基于背侧皮褶窗模型。这些模型通常不允许转移。也有证据表明，肿瘤血管分布可能是器官部位特异性的，因此原位模型将是非常重要的。随着第一阶段资助期间开发的技术，第二阶段资助将进一步开发原位模型中的这些技术，以评估肿瘤血流，血管内肿瘤细胞运输和外渗的抑制剂。

DESCRIPTION (provided by applicant): There is a need for clinically-relevant imageable animal models of tumor blood flow, intravascular cancer-cell trafficking and extravasation, critical targets for current drug evaluation and development. Most models of tumor cell trafficking and blood flow are either based on subcutaneous transplantation of tumors or dorsal skin- fold window models. These models generally do not allow metastasis. There is also evidence that tumor vascularity may be organ-site specific and therefore orthotopic models would be very important. We have previously developed orthotopic metastatic models of cancer which express green fluorescent protein (GFP) (Nature Rev Cancer 5, 796-806, 2005). We have shown that these models enable high resolution whole-body imaging of tumor growth and metastasis and angiogenesis on internal organs (PNAS 97, 1206-11, 2000; 99, 3824-9, 2002). New developments in Phase I include the construction of dual-color cancer cells with GFP in the nucleus and RFP in the cytoplasm (Cancer Res 64, 4251-6, 2004; 65, 4246-52, 2005) that can enable imaging of tumor cell migration, nuclear-cytoplasmic dynamics, and extravasation in the live mouse. Nude mouse models with GFP-expressing blood vessels have also been developed in Phase I which can facilitate imaging of tumor blood flow (Cancer Res 64, 8651-6, 2004; 65, 5352-7, 2005). With these technologies developed during the Phase I grant, the Phase II grant will further develop these technologies in orthotopic models to evaluate inhibitors of tumor blood flow, intravascular tumor cell trafficking and extravasation. The specific aims include: (1) Use of transgenic nude mouse models with GFP blood vessels, orthotopically transplanted with RFP tumor cells for testing agents that target blood flow in the vessels of tumors and their metastasis; (2) Use of dual-color cancer cells with GFP in the nucleus and RFP in the cytoplasm orthotopically implanted in nude mice for in vivo testing of agents that target intravascular trafficking of tumor cells; (3) Use of dual-color cancer cells with GFP in the nucleus and RFP in the cytoplasm orthotopically implanted in nude mice for in vivo testing of agents that target extravasating tumor cells. These orthotopic models are unique in that they will enable visualization tumor blood flow, intravascular tumor-cell trafficking and extravasation at the nuclear-cytoplasmic dynamic level in clinically-relevant mouse models. The newly developed Olympus OV100 whole-mouse imaging system has optics for macro- and high-resolution subcellular imaging along with the models to screen an initial set of potential inhibitors that target these critical steps of metastasis. These models will be used commercially in Phase III to screen and evaluate large numbers of drugs and compound libraries. There is a need for clinically-relevant imageable animal models of tumor blood flow, intravascular cancer-cell trafficking and extravasation, critical targets for current drug evaluation and development. Most models of tumor cell trafficking and blood flow are either based on subcutaneous transplantation of tumors or dorsal skin- fold window models. These models generally do not allow metastasis. There is also evidence that tumor vascularity may be organ-site specific and therefore orthotopic models would be very important. With the technologies developed during the Phase I grant, this Phase II grant will further develop these technologies in orthotopic models to evaluate inhibitors of tumor blood flow, intravascular tumor cell trafficking and extravasation.