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-10 R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”转化为活跃生长的癌症，其表现出向神经周围纤维和淋巴结的局部转移。在该授权申请中，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）以阻断微血管形成、前列腺肿瘤生长和原发性前列腺肿瘤亚系HPCA-10a、10 b、10 c、10 d、10 e和10 f的局部转移。目的1a：表征下调VEGF D合成的IL-10 R轴信号通路;目的1b：评估IL-10受体轴在控制原发性肿瘤细胞系和肿瘤外植体中VEGF D/VEGF C表达中的作用;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。目标二。在原位SCID小鼠模型中测试IL-10转染是否阻断IL-10 S1- VEGF D信号传导以及VEGF D和VEGF C表达以防止淋巴管生成和/或血管生成以及局部转移。目标3：通过免疫测定法评估IL-10 R轴调节的基因/蛋白质的表达，这些基因/蛋白质与人前列腺肿瘤研究中的淋巴管生成（和血管生成）相关。标记研究将使用从大约1294份前列腺癌标本和大约400份活检标本（例如，包括过去5年收集的存档材料）中生成的“组织阵列”块，加上整个前列腺的新鲜冷冻切片进行。我们将比较相对于假定的“潜伏”癌症和恶性癌症以及淋巴结、神经、肌肉和精囊局部转移的病理学的表达谱。将通过使用LYVE-1、Podoplanin和因子VIII抗体进行免疫标记来测量淋巴和血液微血管分布。三维重建成像软件程序ACIS将用于测量连续切片中的微血管密度。对5-10年期间的病理学和患者结果的数据进行评估，将允许评估基因表达谱的重要性。总体而言，这些研究应证明IL-10 R轴和下游效应基因（特别是IL-10 S1-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.