Optical imaging studies of ovarian epithelial cell migration and invasion using microfabricated models of the extracellular matrix

申请者将使用多光子激发(MPE)光化学方法制造卵巢细胞外基质(ECM)模型，作为研究卵巢癌早期阶段的平台。细胞外基质将由交联的基膜和基质层组成，其中每一层都将由体内各自的主要蛋白质组成。由于这些纳米/微结构模型将模拟天然ECM的交联纤维结构，它们将明显比以前报道的(例如培养涂层和流动室)更具仿生性，后者受到缺乏同时适当的地形和生化结构的限制。这些设备将允许对途径活动的测试，这在人类受试者甚至动物模型上是不可能的。这些模型将被用来研究卵巢间质的细胞表面动力学(AIM 1)和细胞外基质重塑(AIM 2)，两者都可能发生在早期癌变中，并可能是疾病进展的基础。为此，他们将从分化、迁移、黏附强度、整合素表达、增殖以及侵袭和重塑细胞外基质的能力等方面比较卵巢肿瘤细胞的功能反应。它们将通过细胞因子和生长因子的刺激，特异性地调节与ECM重塑和转移有关的金属蛋白酶(MMPs)的表达。这些实验将揭示在肿瘤发生和转移中起作用的生化和形态因素。

0853949CampagnolaThe applicants will use multiphoton excited (MPE) photochemistry to fabricate models of the ovarian extracellular matrix (ECM) as platforms to study early stages of ovarian carcinoma. The ECMs will consist of crosslinked basal lamina and stromal layers, where each will be comprised of the respective predominant protein composition in vivo. As these nano/microstructured models will simulate the crosslinked fibrillar structure of the native ECM, they will be significantly more biomimetic than those reported previously (e.g. culture coatings and flow chambers), which are limited by the lack of concurrent appropriate topographic and biochemical structure. These devices will allow the testing of pathway activity not immediately possible in human subjects or even animal models. these models will be used to investigate the cellular surface dynamics (Aim 1) and ECM remodeling of the ovarian stroma (Aim 2) that both may occur in early carcinogenesis and may underlie disease progression. To this end, they will compare the functional response of ovarian tumor cells in terms of differentiation, migration, adhesion strength, integrin expression, proliferation as well as the ability to invade and remodel the ECM. They will specifically modulate the expression of metalloproteinases (MMPs) implicated in ECM remodeling and metastasis via stimulation with cytokines and growth factors. These experiments will reveal the biochemical and morphological factors operative in carcinogenesis and metastasis.