Analysis of the Signaling and Mechanical Cues Promoting Invasion in Melanoma

转移性黑色素瘤是皮肤癌中最致命的一种，也是最具侵袭性的癌症类型之一， 能够迅速扩散出原发部位。该建议旨在分析 黑色素瘤细胞的遗传和信号状态以及细胞微环境的变化， 细胞侵袭性程度和假定的转移潜力。我们的主要假设是 黑色素瘤细胞的侵袭性是由不同因素的组合决定的， 细胞微环境状态的主要贡献，包括ECM的结构和细胞外基质的结构。 可溶性化学因子的存在。细胞微环境的改变，如ECM的排列 纤维，可以允许一个原本非转移性细胞变得更具侵略性，而 表面上转移性细胞的侵袭性可以通过环境变化来调节， 信号级联调节的干预。我们还假设，细胞可以积极地与 不仅通过分泌或降解ECM成分，而且通过积极行使力量， 并因此在最近的锚点（包括脉管系统）的方向上对齐ECM纤维。这 可以反过来产生更多的各向异性的力量，使细胞变形ECM甚至更多， 重新排列的ECM结构向血管迁移的优点。我们将检验这些假设 与ICBP@MIT密切合作，首先利用使用shRNA的新方法， 文库干扰遗传靶标（Aim 1）沿着广泛的环境干扰，然后 使用ICBP（gMIT）提供的最先进的统计分析工具， 数据集，将多种环境和遗传干扰与侵略性细胞行为（目标2）联系起来，我们将 在这个过程中使用几种新的测定方法，增加了数据采集和处理的时间， 以及增强实验的仿生性质。最后，我们将测试由 Aim 2通过一系列测试开发的模型，从细胞培养到组织构建， 模型组织水平（目标3）。我们预计，拟议的研究将揭示新的，以前 环境和遗传因素之间的意外相互作用在控制转移事件， 黑色素瘤，并可能提出对抗这种致命疾病的新方法。结果也可能是 与其他转移性癌症的关系。

Metastatic melanoma is the deadliest form of skin cancer and one of the most aggressive cancer types, capable of rapid spreading out ofthe primary site. This proposal Is aimed at analyzing the combined effect of changes in the genetic and signaling states of melanoma cells and the cell micro-environment on the the degree of cell aggressiveness and putative metastatic potential. The main hypothesis underlying our analysis is that aggressiveness of melanoma cells is determined by a combination of different factors, with a major contribution from the state of cell microenvironment, including the structure ofthe ECM and the presence of soluble chemical factors. Alteration in the cell micro-environment, such as alignment of ECM fibers, can permit an othenwise non-metastatic cell to become more aggressive, whereas the aggressiveness of ostensibly metastatic cell can be moderated by environmental changes and targeted intervention in regulation of signaling cascades. We also hypothesize that cells can actively interact with the micro-environment not only by secreting or degrading ECM components, but by actively exercising forces on and thus aligning ECM fibers in the direction of a nearest anchor point, including the vasculature. This can in turn generate more anisotropic forces allowing cells to deform ECM even more and ultimately take advantage ofthe re-aligned ECM structure to migrate towards blood vessels. We will test these hypotheses in close collaboration with ICBP@MIT, first taking advantage ofthe new methodologies using shRNA libraries to perturb genetic targets (Aim 1) along with a broad range of environmental perturbations and then use the state of the art statistical analysis tools available at ICBP(gMIT to reduce complexity of resulting datasets, linking multiple environmental and genetic perturbation to aggressive cell behavior (Aim 2), We will use several novel assays in the process, increasing manageability of data acquisition and processing, as well as enhancing the biomimetic nature of experimentation. Finally, we will test the predictions arising from the models developed in Aim 2 through a series of tests, ranging from cell culture to tissue construct to model tissue levels (Aim 3). We anticipate that the proposed research will reveal new, previously unanticipated interplay between environmental and genetic factors in control of metastatic events in melanoma and potentially suggest new ways of battling this deadly disease. The results might also have relevance to other metastatic cancers.