Receptor Cross-Talk in Early Metastatic Dissemination

描述（由申请人提供）：转移性传播上皮性卵巢癌（EOC）的受体串扰将影响今天在美国出生的每69名女性中有1名。目前，80%新诊断为EOC的女性已经有转移性疾病，这表明干预转移过程将提高EOC女性的长期生存率。转移是通过一种独特的机制发生的，包括非粘附细胞作为多细胞聚集体（MCAs）脱落到腹膜腔，然后在腹膜内（IP）植入，并且通常与腹膜腹水有关。从游离MCA到危及生命的腹膜锚定转移病变的最终转变的调节因素目前尚不清楚。先前资助期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)-钙粘蛋白（Ecad）表达和功能中的作用。这些机制研究在金属蛋白酶催化的Ecad外结构域脱落、可溶性Ecad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-连环蛋白动力学。此外，我们对原代人EOC中Ecad的表达进行了详细的免疫组化分析，开发了一组用于评估MCA动力学和转移成功的检测方法，并确定了一组通过改变细胞的机械环境来调节的基因产物。这些结果构成了当前假设的基础，即钙粘蛋白转换和IP机械生物学积极促进转移成功。Aim 1的研究将集中在钙粘蛋白转换和MCA动力学上，使用一组钙粘蛋白修饰的细胞系和一套体外试验，旨在机械地评估钙粘蛋白组成对EOC转移中关键细胞事件的影响。目的2的实验将评估改变的腹膜力学生物学对MCA转移成功的一系列措施的影响，并将确定IP机械力是否影响间皮对转移性植入的接受性。目的3将结合人类肿瘤和小鼠IP转移模型的分析，直接检查钙粘蛋白谱的改变、IP力学生物学和体内转移传播。这些研究的长期目标是培养对EOC转移的分子水平理解，这对于开发有效靶向转移性疾病的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. PUBLIC HEALTH RELEVANCE: Epithelial ovarian cancer (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. The long-term goal of these studies is to gain a molecular level understanding of EOC metastasis to design strategies to more effectively target metastatic disease.