Molecular Portraits of Human Breast Cancer Endothelium

几乎所有正在进行临床测试的针对肿瘤新生血管的药物都可以归类为抗肿瘤血管生成药物 血管生成剂。这些药物通常是细胞抑制剂，只抑制正在 增殖、迁移或管状形成，保留已建立的血管。一种替代方法，用于 靶向肿瘤血管包括使用血管靶向剂。这些毒剂本身就有细胞毒性， 理想的靶向是肿瘤内的所有血管，包括新的和已建立的。因为这些化合物是 细胞毒性，重要的是只针对肿瘤内皮细胞，这样就可以避免正常的内皮细胞。因此， 对最具差异性和丰富表达的分子有更详细的了解 肿瘤血管是促进血管靶向药物有效临床开发所必需的。 我们假设乳腺肿瘤内皮细胞在基因表达模式上有质的不同。 来自正常的乳房内皮细胞。我们的目标是获得乳腺肿瘤内皮细胞的分子图谱 并将基因表达模式与正常乳腺内皮细胞进行比较。我们将采用我们的新方法 激光捕获显微切割人、小鼠乳腺肿瘤和人乳腺肿瘤内皮细胞 和小鼠正常乳房组织。我们将提取和扩增RNA，并继续进行基因组图谱 使用44K长的寡点微阵列。我们将着手进行全基因组的基因变化筛查 乳腺肿瘤内皮细胞与正常乳腺组织、腔内皮细胞与正常乳腺组织间的表达 肿瘤的基本亚型。我们还将比较小鼠和人类的基因表达谱 内皮细胞。我们将使用实时荧光定量聚合酶链式反应验证肿瘤内皮标记物的差异表达， 并使用原位杂交或现有抗体来定位转录本或基因产物 来自肿瘤内皮细胞。最后，我们将观察肿瘤内皮细胞的蛋白表达是否发生变化 血管生成治疗转移性乳腺癌患者的临床试验中的标志物 抑制剂2-甲氧基雌二醇和紫杉醇。更详细地了解最重要的分子 在乳腺肿瘤血管上的差异和丰富的表达可能有助于有效的临床 乳腺癌血管靶向药物的开发和诊断标记物的开发 作为抗血管生成治疗反应的替代标志物。

Virtually all drugs targeting tumor neovascularization that are being tested clinically can be classified as anti- angiogenic agents. These agents are generally cytostatic, inhibiting only new blood vessels undergoing proliferation, migration, or tube formation, and sparing established vessels. An alternative approach for targeting tumor vessels involves the use of vascular targeting agents. These agents are inherently cytotoxic, and ideally target all vessels within the tumor, both new and established. Since these compounds are cytotoxic, it is important to target only tumor endothelium, so that normal endothelium are spared. Therefore, a more detailed understanding of the molecules that are most differentially and abundantly expressed on tumor vessels is needed to facilitate the efficient clinical development of vascular targeting agents. We hypothesize that breast tumor endothelium are qualitatively different in their gene expression patterns from normal breast endothelium. Our objective is to obtain molecular profiles of breast tumor endothelium and compare the gene expression patterns to normal breast endothelium. We will employ our novel method for laser capture microdissection of endothelial cells from both human and mouse breast tumors and human and mouse normal breast tissue. We will extract and amplify the RNA, and proceed with genomic profiling using 44k long oligo-spotted microarrays. We will embark on a genome-wide screen for changes in gene expression between endothelium from breast tumors and normal breast tissue, and between luminal and basal subtypes of tumors. We will also compare gene expression profiles between mouse and human endothelium. We will validate the differential expression of tumor endothelial markers using real'time PCR, and use in situ hybridization or existing antibodies to localize transcripts or gene products as being derived from tumor endothelium. Finally, we will see if there is a change in protein expression of tumor endothelial markers in a human clinical trial for patients with metastatic breast cancer treated with the angiogenesis inhibitor 2-methoxyestradiol and paclitaxel. A more detailed understanding of the molecules that are most differentially and abundantly expressed on breast tumor vessels may facilitate the efficient clinical development of vascular targeting agents for breast cancer, and for developing diagnostic markers to serve as surrogate markers for response to anti-angiogenic therapy.