Cell Biological Mechanisms of Melanoma Cell Motility In Vivo

项目概要： 黑色素瘤是最致命的癌症之一，对标准化疗药物的反应很差， 全世界每年有四万八千人死亡近年来，黑色素瘤的发病率急剧上升 几十年大量的研究表明，基质细胞促进肿瘤细胞的运动和转移。 然而，我们对这些相互作用的理解-以及正在开发的基于 根据这种理解-从终点测定和固定的肿瘤切片推断，或基于 在体外共培养模型中以高分辨率可视化细胞。理解肿瘤的一个主要局限 进展是缺乏遗传上易处理的体内模型系统，其适于高分辨率 显像我已经通过可视化和操纵肿瘤细胞及其微环境克服了这一障碍 直接在斑马鱼异种移植模型中。我已经证明了肿瘤和巨噬细胞的相互作用 在哺乳动物系统中观察到的，在人类到斑马鱼的黑色素瘤中得到了非常详细的再现。 异种移植利用一种新的基于Cre重组酶的胞质转移报告基因，我已经证明， 持续的巨噬细胞/肿瘤细胞接触允许细胞质分子从巨噬细胞转移 肿瘤细胞，从而指导肿瘤细胞传播。 我建议使用互补的方法，利用每个生物体的优势来回答 癌症细胞生物学中的一些悬而未决的问题。使用斑马鱼的优势在于， 可以在体内鉴定和验证感兴趣的基因。然后我将使用体外组织培养和 斑马鱼来实时分析细胞间的相互作用。此外，我将使用小鼠模型来测试我们的假设， 哺乳动物体内系统。有了这些方法，我建议确定： 1.什么样的细胞质分子从巨噬细胞转移到肿瘤细胞进行播散？ 2.巨噬细胞如何将细胞质分子转移到肿瘤细胞中进行播散？ 3.来自微环境中基质细胞的哪些额外信号调节肿瘤细胞的细胞骨架 动力学和细胞生物学决策 我的长期目标是了解细胞如何相互沟通，以调节肿瘤细胞的行为， 努力开发新的生物标志物，并确定关键蛋白质和治疗靶向过程。

PROJECT SUMMARY: Melanoma is one of the deadliest forms of cancer and is poorly responsive to standard chemotherapeutics, with 48,000 people dying worldwide each year. The incidence of melanoma has risen dramatically in recent decades. A breadth of work has revealed that stromal cells promote tumor cell motility and metastasis. However, much of our understanding of these interactions - and the therapies that are being developed based on that understanding - is either inferred from end-point assays and fixed tumor sections, or is based on visualizing cells at high resolution in in vitro co-culture models. A major limitation to understanding tumor progression is the lack of genetically tractable in vivo model systems that are amenable to high-resolution imaging. I have overcome this obstacle by visualizing and manipulating tumor cells and their microenvironment directly in a zebrafish xenotransplant model. I have demonstrated that the tumor-macrophage interactions visualized in mammalian systems are recapitulated in great detail in human-to-zebrafish melanoma xenotransplants. With a novel Cre recombinase-based reporter of cytoplasmic transfer, I have shown that sustained macrophage/tumor cell contacts allow for the transfer of cytoplasmic molecules from macrophages to tumor cells, thereby instructing tumor cell dissemination. I propose to use complementary approaches, taking advantage of the strengths of each organism to answer outstanding questions in cancer cell biology. The advantage of using zebrafish is the ease in which genes of interest can be identified and validated in vivo. I will then use a combination of in vitro tissue culture and zebrafish to analyze cellular interactions in real-time. Further, I will use mouse models to test our hypotheses in a mammalian in vivo system. With these approaches in hand, I propose to determine: 1. What cytoplasmic molecules are transferred from macrophages to tumor cells for dissemination? 2. How do macrophages transfer cytoplasmic molecules to tumor cells for dissemination? 3. What additional signals from stromal cells in the microenvironment regulate tumor cell cytoskeletal dynamics and cell biological decision-making? My long term goal is to understand how cells communicate with one another to regulate tumor cell behavior in an effort to develop novel biomarkers, and to identify key proteins and processes to target with therapeutics.