Technology/Map Endothelial Targets/Human Renal Tumors

描述(由申请人提供)： 实体瘤中大多数肿瘤细胞的分子复杂性和体内不可及性极大地限制了基于基因组和蛋白质组的肿瘤特异性成像和治疗药物在体内的有用靶点的发现。为了克服血管内皮细胞(EC)屏障，实现更有效的靶向和穿透实体肿瘤，我们将分析焦点从肿瘤细胞转移到血管EC表面及其与循环血液直接接触的小凹。为了将数据复杂性降低到EC表面表达的有意义的靶向蛋白质子集，我们将使用组织亚细胞分离、新颖的多模质谱分析、电子消减以及结构和功能的生物信息学查询，从组织中的100,000个蛋白质中揭示出在人类肾脏肿瘤的血管内皮细胞上差异表达的少数静脉可获得的蛋白质。这种技术和整体方法已经在啮齿动物实体肿瘤中得到验证，新的血管靶点被发现，从而允许肿瘤特异性成像、穿透和有效的放射免疫治疗(自然，429：629-35,2004)。但是，目前对肿瘤新生血管内皮细胞，尤其是人体组织中蛋白质的表达知之甚少。现在，我们希望应用我们的新技术，在活体内全面绘制人类肾脏肿瘤中腔内皮细胞表面和小凹的蛋白质组图。人类肿瘤很可能会在肿瘤新生血管上表达一系列不同的蛋白质，这些蛋白质在动物模型中尚未发现或诱导。为此，我们提出了以下具体目标：1)使用新的组织亚分离和蛋白质组学分析方法，全面定位人肾肿瘤与匹配的正常肾组织中的血管EC表面和小窝，以揭示肿瘤诱导/相关的候选血管蛋白。2)创造针对新发现的人肾肿瘤EC靶点的新抗体，并以抗体为探针，验证肿瘤诱导/相关蛋白在人体组织EC表面及其小凹的表达，从而评价人实体肿瘤新生血管的靶向特异性程度。这种图谱还可以阐明肿瘤对发育中的血管内皮细胞的影响，产生重要的肿瘤特异性血管靶点，用于改进非侵入性诊断成像和治疗，并为肿瘤活检的分子分类产生新的诊断和预后标志。

DESCRIPTION (provided by applicant): The molecular complexity and in vivo inaccessibility of most tumor cells within solid tumors can greatly limit genomic- and proteomic-based discovery of useful targets for tumor-specific imaging and therapeutic agents in vivo. To overcome endothelial cell (EC) barriers and achieve more effective targeting and penetration into solid tumors, we shift analytical focus from the tumor cell to the vascular EC surface and its caveolae in direct contact with the circulating blood. To reduce data complexity to a meaningful subset of targetable proteins expressed on the EC surface, we will use tissue sub-cellular fractionation, novel multimodal mass spectrometric analysis, in silico subtraction, and bioinformatics interrogation of structure and function to unmask, from the >100,000 proteins in the tissue, those few intravenously accessible proteins differentially expressed on vascular endothelium in human renal tumors. This technology and overall approach has been validated in rodent solid tumors whereby new vascular targets have been uncovered permitting tumor-specific imaging, penetration, and effective radio immunotherapy (Nature, 429:629-35, 2004). But, currently very little is known about the expression of proteins in tumor neovascular endothelium, especially in human tissue. We now wish to apply our new technology to map comprehensively the proteome of luminal EC surfaces and caveolae in human renal tumors in vivo. It is likely that human tumors will express a different constellation of proteins on tumor neovasculature not yet uncovered or induced in animal models. To this end, we propose the following specific aims: 1) To use novel tissue sub fractionation and proteomic analytical approaches to map comprehensively vascular EC surfaces and caveolae in human renal tumors vs. matched normal renal tissue to unmask candidate tumor-induced/associated vascular proteins. 2) To create new antibodies to newly discovered human renal tumor EC targets and to use antibodies as probes to validate the expression of tumor-induced/associated proteins at the EC surface and its caveolae in human tissues and thereby to assess the degree of target specificity for the neovasculature of human solid tumors. Such mapping may also elucidate the effects of the tumor on the developing vascular endothelium and yield important tumor-specific vascular targets for improving noninvasive diagnostic imaging and therapy as well as yield new diagnostic and prognostic markers for the molecular classification of tumor biopsies