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形式，也是最具侵袭性的癌症类型之一。 能够迅速扩散到主要场所之外。这项建议旨在分析联合效应。 黑色素瘤细胞的遗传和信号状态以及细胞微环境的变化 细胞侵袭程度和可能的转移潜能。我们背后的主要假设是 分析认为，黑色素瘤细胞的侵袭性是由不同因素的组合决定的，具有 细胞微环境状态的主要贡献，包括细胞外基质的结构和 存在可溶的化学因素。细胞微环境的改变，如细胞外基质的排列 纤维可以使非转移性细胞变得更具侵袭性，而 表面转移细胞的侵袭性可以通过环境变化和靶向来缓和 干预信号级联的调节。我们还假设细胞可以主动地与 微环境不仅通过分泌或降解ECM组件，而且通过积极行使力量 使ECM纤维沿着最近的锚定点(包括血管系统)的方向排列。这 可以反过来产生更多的各向异性力，使细胞更多地变形ECM并最终 重新对齐的ECM结构的优势是向血管迁移。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 扰乱遗传目标的文库(目标1)以及广泛的环境扰动，然后 使用ICBP(GMIT)提供的最先进的统计分析工具来降低结果的复杂性 数据集，将多个环境和遗传扰动与侵略性细胞行为联系起来(目标2)，我们将 在该过程中使用几种新的分析方法，提高了数据采集和处理的可管理性，如 以及增强实验的仿生性。最后，我们将测试由以下因素引起的预测 在AIM 2中通过一系列测试开发了模型，从细胞培养到组织构建，再到 模型组织水平(目标3)。我们预计，拟议的研究将揭示新的、以前的 环境和遗传因素在控制肿瘤转移事件中的意外相互作用 黑色素瘤，并可能提出与这种致命疾病作斗争的新方法。结果可能还会有 与其他转移性癌症的相关性。

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.