Color-Coded Imaging of Pancreatic Cancer Microenvironment for Drug Discovery

描述(由申请人提供)：我们建议加州大学圣迭戈分校(学术伙伴)和抗癌(工业伙伴)的研究人员建立独特的学术-产业合作伙伴关系，在胰腺癌小鼠模型中开发和验证颜色编码荧光成像系统。这项拟议的研究将促进用于老鼠模型研究的先进成像技术、方法和工具。这些研究的发现将促进我们对胰腺癌中肿瘤-宿主相互作用的了解，并将导致这种几乎一致致命的疾病的新疗法。假说彩色编码荧光成像可以区分胰腺癌细胞和宿主血管及间质细胞，这将有助于评价选择性抗间质药物。具体目标1(1-3岁)发展胰腺癌微环境的颜色编码可成像模型。我们开发了一种简单但功能强大的新模型，用于使用双色荧光成像来描绘肿瘤诱导的血管生成的形态事件。宿主模型为本实验室研制的ND-GFP转基因裸鼠，新生血管用GFP标记，肿瘤用红色荧光蛋白(RFP)标记。我们计划用从无处不在表达GFP、RFP、CFP(蓝色)和GFP-RFP(黄色)转基因免疫活性小鼠获得的基质细胞(巨噬细胞、淋巴细胞、树突状细胞和骨髓细胞)对该模型的肿瘤微环境进行颜色编码。具体目标2(2年至4年)开发一种模型系统，允许对小鼠淋巴管和淋巴管内癌细胞的运输进行实时体内成像，以促进转移性肿瘤细胞的淋巴靶向研究。我们正在开发一种模型系统，可以对小鼠淋巴管和淋巴管内癌细胞的转运进行实时体内成像，以促进转移性肿瘤细胞的淋巴靶向研究。该模型利用RFP标记的人胰腺癌细胞和抗LYVE1的单抗标记淋巴管，以成像肿瘤细胞的淋巴传输。输送到淋巴系统的细胞被实时记录在视频中，离开腹股沟淋巴结，进入腋窝淋巴结。使用荧光LYVE-1可以对肿瘤细胞与淋巴管壁和瓣膜的相互作用进行详细的成像，便于观察淋巴管内细胞运输的动态。具体目标3(3-5年)在我们的颜色编码的原位和淋巴小鼠模型中确定新的抗间质疗法治疗胰腺癌的有效性。几种新的抗间质、抗血管生成和抗淋巴管生成药物，包括抗整合素的单抗和小分子血管生成抑制剂，将在特定AIMS 1和2中开发的新的颜色编码胰腺癌微环境模型中进行有效性测试。这些颜色编码的人胰腺癌裸鼠模型将用于可视化新的靶点，这将极大地促进胰腺癌抗基质、抗血管生成和抗淋巴管生成药物的发现。

DESCRIPTION (provided by applicant): We propose a unique academic-industrial partnership between investigators at UCSD (Academic Partner) and AntiCancer (Industrial Partner) to develop and validate color-coded fluorescent imaging systems in mouse models of pancreatic cancer. The proposed research will facilitate advanced imaging technology, methods and tools for mouse-model studies. Findings from these studies will advance our knowledge of tumor- host interactions in pancreatic cancer and will lead to novel treatments for this almost uniformly fatal disease. Hypothesis Color-coded fluorescence imaging can distinguish pancreatic cancer cells from host angiogenic vessels and stromal cells and will be useful for evaluating selective anti-stromal agents. Specific Aim 1 (Years 1-3) Development of color-coded imageable models of the pancreatic cancer microenvironment. We have developed a simple yet powerful new model for delineating the morphological events of tumor-induced angiogenesis using dual color fluorescence imaging. The host model is ND-GFP transgenic nude mouse, developed in our laboratory, in which nascent blood vessels are labeled with GFP and tumors are labeled with red fluorescent protein (RFP). We plan to color-code the tumor microenvironment of this model with stromal cells (macrophages, lymphocytes, dendritic, and bone marrow cells) harvested from ubiquitously-expressing GFP, RFP, CFP (blue) and GFP-RFP (yellow) transgenic immunocompetent mice. Specific Aim 2 (Years 2-4) Development of a model system allowing real-time in-vivo imaging of murine lymphatics and intralymphatic cancer cell trafficking to facilitate the study of lymph node targeting by metastatic tumor cells. We are developing a model system allowing real-time in-vivo imaging of murine lymphatics and intralymphatic cancer cell trafficking to facilitate the study of lymph node targeting by metastatic tumor cells. The model utilizes RFP-labeled human pancreatic cancer cells and monoclonal anti-LYVE1 antibody to label lymphatic vessels in order to image lymphatic trafficking of tumor cells. Cells delivered to the lymphatic system are recorded on video in real-time leaving the inguinal lymph node and entering the axillary lymph node. The use of fluorescent LYVE-1 allows detailed imaging of tumor cell interaction with lymphatic vessel walls and valves, facilitating observation of the dynamics of intralymphatic cellular trafficking. Specific Aim 3 (Years 3-5) Determination of the efficacy of novel anti-stromal therapies for pancreatic cancer in our color-coded fluorescent in vivo orthotopic and lymphatic mouse models. Several novel anti-stromal, anti-angiogenic, and anti-lymphangiogenic agents including monoclonal antibodies against integrins and small molecule inhibitors of angiogenesis will be tested for their efficacy in the novel color-coded pancreatic cancer microenvironment models developed in specific aims 1 and 2. These color-coded nude mouse models of human pancreatic cancer will be used to visualize new targets that should greatly enhance the discovery of anti-stromal, anti-angiogenic and anti-lymphangiogenic drugs for pancreatic cancer.