Receptor Cross-Talk in Early Metastatic Dissemination

描述(由申请人提供)：转移性播散性上皮性卵巢癌(EOC)的受体串扰将影响今天在美国出生的每69名妇女中的1名。目前，80%的新诊断为EoC的妇女已经有转移疾病，这表明对转移过程的干预将提高EoC妇女的长期存活率。肿瘤转移是通过一种独特的机制发生的，这种机制涉及非黏附细胞以多细胞聚集体(MCAS)的形式脱落到腹膜腔内，然后在腹膜内植入(IP)，通常与腹水相关。调节从自由漂浮的大脑中动脉到危及生命的腹膜锚定转移性病变的终末转变的因素目前尚不清楚。上一次资助期间的研究强调了可溶性微环境调节剂EGF和溶血磷脂酸(LPA)在调节上皮(E)-钙粘附素(ECAD)表达和功能中的作用。这些机制研究产生了令人兴奋的新数据，关于金属蛋白酶催化的ECAD胞外结构域脱落，可溶性ECAD胞外结构域在人EOC腹水中的作用，以及连接完整性改变导致的β-连环蛋白动力学的变化。此外，我们对原代EOC中ECAD的表达进行了详细的IHC分析，开发了一组用于评估MCA动力学和转移成功的分析方法，并确定了一组通过改变细胞机械环境来调节的基因产物。这些结果构成了目前的假设的基础，即钙粘蛋白转换和IP机制生物学积极地促进了转移成功。目标1的研究将集中于钙粘蛋白转换和大脑中动脉动力学，使用一组钙粘素修饰的细胞系和一套体外测试，旨在从机械上评估钙粘素成分对EOC转移中关键细胞事件的影响。Aim 2中的实验将评估腹膜机械生物学改变对MCA转移成功的一系列测量的影响，并将确定IP机械力是否影响间皮细胞对转移种植的容受性。目的3将结合对人类肿瘤和小鼠IP转移模型的分析，直接检查改变的钙粘素分布、IP机制生物学和体内转移扩散。这些研究的长期目标是培养对EOC转移的分子水平的了解，这对于开发有效针对转移性疾病的EOC特异性治疗是必要的。

DESCRIPTION (provided by applicant): Receptor Cross-Talk in Metastatic Dissemination Epithelial ovarian carcinoma (EOC) will affect 1 out of every 69 women born in the US today. Currently, 80% of women newly diagnosed with EOC already have metastatic disease, indicating that intervention in the metastatic process will improve long-term survival of women with EOC. Metastasis occurs through a unique mechanism involving shedding of non-adherent cells as multi-cellular aggregates (MCAs) into the peritoneal cavity followed by intra-peritoneal (IP) implantation, and is often associated with peritoneal ascites. The factors that regulate the terminal transition from free-floating MCA to life-threatening peritoneally anchored metastatic lesion are currently unknown. Studies in the previous funding period highlighted the role of the soluble microenvironmental regulators EGF and lysophosphatidic acid (LPA) in regulation of epithelial (E)-cadherin (Ecad) expression and function. These mechanistic studies generated exciting new data on metalloproteinase-catalyzed Ecad ectodomain shedding, the role of the soluble Ecad ectodomain in human EOC ascites, and on changes in ?-catenin dynamics resulting from altered junctional integrity. Further, we performed a detailed IHC analysis of Ecad expression in primary human EOC, developed a panel of assays with which to evaluate MCA dynamics and metastatic success, and identified a set of gene products regulated by altering the mechanical environment of the cell. These results form the basis of the current hypothesis that cadherin switching and IP mechanobiology actively contribute to metastatic success. Studies in Aim 1 will focus on cadherin switching and MCA dynamics using a panel of cadherin-modified cell lines and a suite of in vitro assays designed to mechanistically evaluate the effect of cadherin composition on key cellular events in EOC metastasis. Experiments in Aim 2 will evaluate the effect of altered peritoneal mechanobiology on the suite of measures of MCA metastatic success and will determine whether IP mechanical forces affect mesothelial receptivity to metastatic implantation. Aim 3 will combine analysis of human tumors and murine IP metastasis models for a direct examination of altered cadherin profiles, IP mechanobiology, and metastatic dissemination in vivo. The long-term goal of these studies is to cultivate a molecular level understanding of EOC metastasis, necessary for the development of EOC-specific therapies that effectively target metastatic disease.