IL-10 Regulation of Lymphangiogenesis in Prostate Cancer

描述(申请人提供)：根据初步研究，我们推测，IL-10/IL-10R轴控制着导致微血管形成和肿瘤从“潜伏癌”转变为活跃生长癌症的“淋巴管生成开关”，后者表现为局部转移到神经周纤维和淋巴结。在这项赠款申请中，总的目标是评估IL-10是否选择性地靶向淋巴管生成(和/或血管生成)，以阻止原发前列腺癌亚系HPCA-10a、10b、10c、10d、10e和10f的微血管形成、前列腺癌生长和局部转移。目的1a：研究下调VEGFD合成的IL-10R轴信号通路；目的1b：评价IL-10受体在原代肿瘤细胞株和肿瘤组织中调控VEGFD/VEGFC表达的作用；目的1c：确定IL-10对体外肿瘤细胞诱导的淋巴管生成(和血管生成)的影响。目的2.在原位SCID小鼠模型中，检测IL-10能否阻断IL-10S1-VEGFD信号转导及VEGFD和VEGFC的表达，从而阻止淋巴管生成和/或血管生成以及局部转移。目的：用免疫分析方法研究人前列腺癌中与淋巴管生成(和血管生成)相关的受IL-10R轴调控的基因/蛋白的表达。标记研究将使用从大约1294个前列腺癌样本和大约400个活检组织(例如，包括过去5年收集的档案材料)产生的‘组织阵列’块，以及新鲜的整个前列腺癌的冰冻切片进行。我们将比较与假定的“潜伏性”癌症、恶性肿瘤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。淋巴管和血管微血管分布将用LYVE-1、泊多普宁和因子抗体进行免疫标记。一个名为ACIS的三维重建成像软件程序将被用来测量连续切片的微血管密度。对5-10年的病理和患者转归的数据进行评估，可以评估基因表达谱的重要性。总之，这些研究应该证明IL-10R轴和下游效应基因(特别是IL-10S1-VEGFD和VEGFD)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.