IL-10 Regulation of Lymphangiogenesis in Prostate Cancer

描述（由申请人提供）：基于初步研究，我们假设IL-10/IL-10R轴控制“淋巴管生成开关”，导致微血管形成和肿瘤从“潜伏性癌症”转变为活跃生长的癌症，并表现出局部转移到神经周围纤维和淋巴结。在这项拨款申请中，总体目标是评估IL-10是否选择性地靶向淋巴管生成（和/或血管生成），以阻断原发性前列腺肿瘤亚群HPCA-10a、10b、10c、10d、10e和10f的微血管形成、前列腺肿瘤生长和局部转移。目标有三个，包括：目的1a：表征IL-10R轴信号通路，其下调VEGF D的合成；目的1b：探讨IL-10受体轴在原代肿瘤细胞系和肿瘤外植体中调控VEGF D/VEGF C表达的作用；目的：探讨IL-10对体外肿瘤细胞诱导的淋巴管生成（及血管生成）的影响。目标2。在原位SCID小鼠模型中检测IL-10转染是否阻断IL-10S1- VEGF D信号和VEGF D和VEGF C的表达以阻止淋巴管生成和/或血管生成，以及局部转移。目的3：通过免疫分析评估IL-10R轴调控的基因/蛋白的表达，这些基因/蛋白在人类前列腺肿瘤研究中与淋巴管生成（和血管生成）相关。标记研究将使用“组织阵列”块进行，这些块来自大约1294例前列腺癌标本和大约400例活组织切片（例如，包括过去5年收集的档案材料），以及整个前列腺的新鲜冷冻切片。我们将比较与假定的“潜伏性”癌症、恶性癌症以及局部转移到淋巴结、神经、肌肉和精囊的病理相关的表达谱。淋巴和微血管分布将通过LYVE-1、Podoplanin和Factor VIII抗体的免疫标记来测量。一个三维重建成像软件程序称为ACIS将用于测量微血管密度在连续切片。对5-10年期间的病理和患者预后数据的评估将允许评估基因表达谱的重要性。总的来说，这些研究应该证明IL-10R轴和下游效应基因（特别是IL-10S1-VEGF D和VEGF D）是否在淋巴管生成和/或血管生成过程中发挥关键作用。

DESCRIPTION (provided by applicant): We postulate, based on preliminary studies, that the IL-10/IL-10R axis controls a 'lymphangiogenesis switch' leading to microvessel formation and the transformation of tumors from 'latent cancers' to actively growing cancers, which exhibit localized metastases to the perineural fibers and lymph nodes.. In this grant application, the overall goal is to assess whether IL-10 selectively targets lymphangiogenesis (and/or angiogenesis) to block microvessel formation, prostate tumor growth and localized metastases by primary prostate tumor sublines, HPCA-10a, 10b, 10c, 10d, 10e and 10f. The goals are three foal, including: Aim 1a: to characterize the IL-10R axis signaling pathway, which down regulates VEGF D synthesis; Aim 1b: to evaluate the role of the IL-10 receptor axis in controlling VEGF D/VEGF C expression in primary tumor cell lines and tumor explants; Aim lc: to determine the influence of IL-10 on tumor cell induced lymphangiogenesis (and angiogenesis) in vitro. Aim 2. To test whether IL-10 transfection blocks IL-10S1- VEGF D signaling and VEGF D and VEGF C expression to prevent lymphangiogenesis and/or angiogenesis, and localized metastases in an orthotopic SCID mouse model. Aim 3: to evaluate the expression by immunoassays of the genes/proteins regulated by the IL-10R axis, which are associated with lymphangiogenesis (and angiogenesis) in human prostate tumor studies. The labeling studies will be done with 'tissue arrays' blocks generated from approximately 1294 prostate cancer specimens and approximately 400 biopsies (e.g. including archival material collected over the past 5 yrs), plus fresh frozen sections of whole mount prostates. We will compare the expression profiles relative to the pathology of putative 'latent' cancers, and malignant cancers as well as localized metastases to the lymph nodes, neural, muscle, and seminal vesicles. Lymphatic and blood microvessel distributions will be measured by immunolabeling with LYVE-1, Podoplanin and Factor VIII antibodies. A 3-D reconstruction imaging software program termed ACIS will be employed for measuring microvessel density in serial sections. Evaluation of the data with respect to the pathology and patient outcome over a 5-10 yr period will permit evaluation of the importance of gene expression profiles. Overall, the studies should demonstrate whether the IL-10R axis and down stream effector genes (and specifically IL-10S1-VEGF D, and VEGF D) play a pivotal role in lymphangiogenesis and/or angiogenesis processes.