Nanoparticle Imaging and Co-Targeting of Cancer and Bone Stroma

项目6将使用纳米载体来成像和共同靶向致命的前列腺癌和骨转移。这 该提案建立在我们实验室利用生物发光跟踪正常人的有希望的初步研究基础上。 转基因小鼠中的前列腺上皮细胞和前列腺特异性膜抗原（PSMA）抗体 共轭纳米粒子跟踪活体动物前列腺肿瘤。在本申请中，我们进一步提出， 开发和应用尖端纳米技术来成像和靶向骨转移，这是一种致命的癌症 前列腺癌、乳腺癌和肺癌患者的表型。主要目标是开发多功能 用于成像、靶向和监测细胞培养物和动物中前列腺癌细胞反应的试剂 模型我们还建议建立一种单细胞间隙连接染料转移法来区分前列腺 具有不同恶性程度的癌细胞。提出了四个具体目标。具体目标1将 在临床人前列腺中验证前列腺癌、骨基质和内皮细胞表面生物标志物 用于成像和靶向的癌症标本。我们将确认细胞表面蛋白的表达 快速尸检骨转移标本中的生物标志物，并开发间隙连接染料转移 用于前列腺癌和前列腺或骨基质细胞的测定。具体目标2将探索细胞的使用 表面生物标记物作为靶向模块，用于开发前列腺癌、骨 基质细胞和内皮细胞作为3-D嵌合前列腺癌类器官生长，并在转基因、原位和 异种移植小鼠模型。将使用用以下标记的细胞校准测定的定量方面： 荧光蛋白，使用萤火虫荧光素酶的生物发光成像将在培养的细胞中进行测试， 小鼠模型。类似的方案将用于靶向前列腺癌、骨基质和内皮细胞 具体目标3。我们将开发一种成像方法来检测凋亡细胞膜联蛋白V成像。 将合成基于肝素的可生物降解的紫杉醇缀合物，并在细胞培养和动物实验中进行测试。 模型具体目标4将构建多功能纳米颗粒，以同时成像，靶向， 在动物模型中监测前列腺癌、骨和淋巴结转移的治疗反应。 最终，该项目将开发新的方法和多功能试剂来成像，靶向和监测 治疗前列腺癌骨和淋巴结转移的治疗反应。

Project 6 will use nanovectors to image and cotarget lethal prostate cancer and bone metastasis. This proposal builds on promising preliminary studies from our laboratory using bioluminescence to track normal prostate epithelial cells in transgenic mice and prostate-specific membrane antigen (PSMA) antibody conjugated nanoparticles to track prostate tumors in live animals. In this application, we further propose to develop and apply cutting edge nanotechnology to image and target bone metastasis, a lethal cancer phenotype in prostate, breast, and lung cancer patients. The primary goal is to develop multifunctional reagents to image, target and monitor the response of prostate cancer cells in cell culture and animal models. We also propose to develop a single cell gap junction dye transfer assay to differentiate prostate cancer cells with varying degrees of malignancy. Four specific aims are proposed. Specific aim 1 will validate prostate cancer, bone stromal, and endothelial cell surface biomarkers in clinical human prostate cancer specimens for imaging and targeting. We will confirm the expression of cell surface protein biomarkers in bone metastatic specimens from rapid autopsies and develop a gap junction dye transfer assay for prostate cancer and prostate or bone stromal cells. Specific aim 2 will explore the use of cell surface biomarkers as a targeting module to develop molecular imaging methods for prostate cancer, bone stromal, and endothelial cells grown as 3-D chimeric prostate cancer organoids and in transgenic, in situ and xenograft mouse models. Quantitative aspects of the assay will be calibrated using cells tagged with fluorescent proteins, Bioluminescent imaging using firefly luciferase will be tested in cultured cells and mouse models. Similar protocols will be used to target prostate cancer, bone stromal, and endothelial cells as in specific aim 3. We will develop an imaging method to detect apoptotic cells using annexin V imaging. Heparin-based biodegradable paclitaxel conjugates will be synthesized and tested in cell culture and animal models. Specific aim 4 will construct multifunctional nanoparticles to simultaneously image, target, and monitor therapeutic responses of prostate cancer, bone and lymph node metastases in animal models. Ultimately, this project will develop novel methods and multifunctional reagents to image, target, and monitor therapeutic responses to treat men with prostate cancer bone and lymph node metastases.