Nanoparticles for Harvesting and Targeting Angiogenic Proteins

描述(由申请人提供)：本R21/R33申请名为“用于采集和靶向血管生成蛋白的纳米颗粒”，其假设是，开发和完善具有纳米特性的硅芯片的表面特征可以提高对血清和肿瘤中存在的低分子血管生成蛋白的质谱仪(MS)检测的灵敏度，这些低分子血管生成蛋白在肿瘤发展的非常早期就产生了。此外，改进纳米多孔颗粒的偶联方法将允许选择性靶向体外的内皮细胞和体内的肿瘤相关血管，并结合负载策略的改进，负载到纳米颗粒中的细胞毒剂可以选择性地破坏这些血管。我们的实验计划基于我们在开发和改进新兴纳米技术方法方面的专业知识，这些方法用于捕获蛋白质、选择性靶向和加载硅纳米颗粒。这些研究还利用了我们在鉴定血管内皮生长因子(VEGF)蛋白家族中的新蛋白方面的经验，这些蛋白在肿瘤相关血管生成过程中是必不可少的。为了实现开发和改进检测血管生成蛋白和选择性靶向和破坏肿瘤相关血管的工具的目标，提出了以下具体目标：1.开发和精制具有纳米特征的硅胶芯片，以提高在体内皮肤肿瘤相关血管生成过程中血清和皮肤肿瘤中VEGF蛋白的LC-MS/MS鉴定的灵敏度；2.改进硅纳米颗粒与抗VEGFR-2受体抗体的偶联，以选择性地体外靶向内皮细胞和体内靶向肿瘤相关血管；3.测定与抗VEGFR-2抗体偶联的硅纳米粒负载和传递细胞毒剂Melatin的能力，以在体外破坏内皮细胞，在体内破坏肿瘤相关血管。这些研究将提供敏感的纳米技术工具，在确定血清和肿瘤中与肿瘤血管生成相关的蛋白质组方面至关重要，这一点目前尚未被探索。这些研究还可能提供使用纳米技术方法选择性地针对肿瘤血管进行破坏的策略。

DESCRIPTION (provided by applicant): This R21/R33 application entitled "Nanoparticles for Harvesting and Targeting Angiogenic Proteins" has as its hypothesis that development and refinement of surface characteristics of silica chips with nanocharacteristics can enhance sensitivity of mass spectrometry (MS) detection of the low molecular weight angiogenic proteins present in serum and tumors that produced at very early times of tumor development. In addition, refinement of conjugation methods of nanoporous particles will allow selective targeting of endothelial cells in vitro and tumor-associated blood vessels in vivo and in combination with refinement of loading strategies, cytotoxic agents loaded into nanoparticles can selectively destroy these vessels. Our experimental plan is based on our expertise in development and refinement of emerging nanotechnology approaches for protein capture, for selective targeting and loading of silicon nanoparticles. These studies also take advantage of our experience in identification of novel proteins within the vascular endothelial growth factor (VEGF) family of proteins that are essential in the process of tumor-associated angiogenesis. To achieve the goal of developing and refining tools for detection of angiogenic proteins and for selective targeting and destruction of tumor-associated blood vessels, the following Specific Aims are proposed: 1. Develop and refine silica chips with nanocharacteristics to enhance the sensitivity of LC-MS/MS identification VEGF proteins in serum and in skin tumors during skin tumor-associated angiogenesis in vivo; 2. Refine conjugation of silicon nanoparticles to anti-VEGFR-2 receptor antibodies for selective targeting of endothelial cells in vitro and targeting tumor-associated blood vessels in vivo; 3. Determine the ability of silicon nanoparticles conjugated with anti-VEGFR-2 antibodies to be loaded with and to deliver the cytotoxic agent melatin for destruction of endothelial cells in vitro and for destruction of tumor-associated blood vessels in vivo. These studies will provide sensitive nanotechnology tools that are critical in defining the proteome in serum and tumors related to tumor angiogenesis that is currently unexplored. These studies may also provide strategies to selectively target tumor vessels for destruction using nanotechnology approaches.