Color-Coded Imaging of Pancreatic Cancer Microenvironment for Drug Discovery

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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.