Color-Coded Imaging of Pancreatic Cancer Microenvironment for Drug Discovery

描述（由申请人提供）：我们提议UCSD（学术合作伙伴）和AntiCancer（工业合作伙伴）的研究人员之间建立独特的学术-工业合作伙伴关系，以开发和验证胰腺癌小鼠模型中的颜色编码荧光成像系统。拟议的研究将促进先进的成像技术，方法和工具的小鼠模型研究。这些研究的发现将推进我们对胰腺癌中肿瘤-宿主相互作用的认识，并将为这种几乎一致致命的疾病带来新的治疗方法。假设彩色编码荧光成像可以区分胰腺癌细胞从宿主血管生成血管和基质细胞，并将用于评估选择性抗基质剂。具体目标1（1-3年级）开发胰腺癌微环境的彩色编码成像模型。我们已经开发了一个简单而强大的新模型，用于描绘肿瘤诱导的血管生成的形态学事件，使用双色荧光成像。宿主模型是本实验室开发的ND-GFP转基因裸鼠，其中新生血管用GFP标记，肿瘤用红色荧光蛋白（RFP）标记。我们计划用从普遍表达GFP、RFP、CFP（蓝色）和GFP-RFP（黄色）转基因免疫活性小鼠中收获的基质细胞（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（第2-4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运输进行实时体内成像，以促进转移性肿瘤细胞对淋巴结靶向的研究。我们正在开发一个模型系统，允许实时在体内成像的小鼠淋巴管和淋巴管内癌细胞的运输，以促进淋巴结靶向转移性肿瘤细胞的研究。该模型利用RFP标记的人胰腺癌细胞和单克隆抗LYVE 1抗体标记淋巴管，以成像肿瘤细胞的淋巴运输。输送到淋巴系统的细胞在视频上实时记录离开腹股沟淋巴结和进入腋窝淋巴结。荧光LYVE-1的使用允许肿瘤细胞与淋巴管壁和瓣膜相互作用的详细成像，便于观察淋巴管内细胞运输的动力学。具体目标3（第3-5年）在我们的彩色编码荧光体内原位和淋巴小鼠模型中确定新型抗间质疗法对胰腺癌的疗效。将检测几种新型抗基质、抗血管生成和抗淋巴管生成药物（包括抗整联蛋白的单克隆抗体和血管生成小分子抑制剂）在特定目标1和2中开发的新型彩色编码胰腺癌微环境模型中的疗效。这些彩色编码的人胰腺癌裸鼠模型将被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。公共卫生相关性：胰腺癌是一种致命性疾病，5年生存率仅为1- 4%。显然，需要新的治疗策略。我们建议在加州圣地亚哥大学（学术合作伙伴）和抗癌公司的研究人员之间建立一个独特的学术产业伙伴关系。（工业合作伙伴）开发和验证可用于评估新药的胰腺癌小鼠模型中的颜色编码荧光成像系统。

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. PUBLIC HEALTH RELEVANCE: Pancreatic cancer is a fatal disease with 5-year survival rates of only 1-4%. Clearly, new treatment strategies are needed. We propose a unique academic industrial partnership between investigators at the University of California San Diego (Academic Partner) and AntiCancer, Inc. (Industrial Partner) to develop and validate color-coded fluorescent imaging system in mouse models of pancreatic cancer that can be used for evaluation of new drugs.