Cell Biological Mechanisms of Melanoma Cell Motility In Vivo

体内黑色素瘤细胞运动的细胞生物学机制

• 批准号: 9792230
• 负责人: Minna Roh
• 金额: $ 9.5万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9792230
• 关键词: Biochemical; Biological; Biological Models; Biological Process; Blood Vessels; Cell Communication; Cell Culture System; Cell physiology; Cells; Cellular Structures; Cellular biology; Clinic; Coculture Techniques; Cytoplasm; Decision Making; Disease; Drug Delivery Systems; Drug Targeting; Electron Microscopy; End Point Assay; Enterobacteria phage P1 Cre recombinase; Fibroblasts; Genes; Genetic; Goals; Hand; Human; Immune; In Vitro; Incidence; Infiltration; Malignant Neoplasms; Melanoma Cell; Microscopy; Modeling; Mole the mammal; Molecular; Mus; Nature; Neoplasm Metastasis; Organism; Pharmaceutical Preparations; Phase; Primary Neoplasm; Process; Proteins; Reporter; Resolution; Role; Signal Transduction; Stromal Cells; Structure; System; Testing; Therapeutic; Time; Tumor Cell Invasion; Tumor-associated macrophages; Vascularization; Work; Xenograft procedure; Zebrafish; base; cancer cell; cell behavior; cell motility; experimental study; high resolution imaging; in vivo; in vivo Model; interest; light microscopy; macrophage; melanoma; mouse model; neoplastic cell; neutrophil; novel; novel marker; outcome forecast; tissue culture; tumor; tumor initiation; tumor microenvironment; tumor progression

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.

项目概要： 黑色素瘤是最致命的癌症之一，对标准化疗反应不佳， 全球每年有 48,000 人死亡。近年来黑色素瘤的发病率急剧上升 几十年。大量的工作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以及正在开发的疗法的大部分理解是基于 基于这种理解 - 要么从终点测定和固定肿瘤切片推断，要么基于 在体外共培养模型中以高分辨率可视化细胞。了解肿瘤的主要限制 进展是缺乏适合高分辨率的遗传易处理的体内模型系统 成像。我通过可视化和操纵肿瘤细胞及其微环境克服了这个障碍 直接在斑马鱼异种移植模型中进行。我已经证明肿瘤与巨噬细胞的相互作用 在哺乳动物系统中可视化的现象在人类到斑马鱼的黑色素瘤中得到了非常详细的重现 异种移植。通过一种新型的基于 Cre 重组酶的细胞质转移报告基因，我已经证明 持续的巨噬细胞/肿瘤细胞接触允许巨噬细胞转移细胞质分子 作用于肿瘤细胞，从而指导肿瘤细胞扩散。 我建议使用互补的方法，利用每个有机体的优势来回答 癌细胞生物学中的突出问题。使用斑马鱼的优点是很容易将 可以在体内识别和验证兴趣。然后我将结合使用体外组织培养和 斑马鱼实时分析细胞相互作用。此外，我将使用小鼠模型来检验我们的假设 哺乳动物体内系统。有了这些方法，我建议确定： 1. 哪些细胞质分子从巨噬细胞转移到肿瘤细胞进行传播？ 2.巨噬细胞如何将胞浆分子转移至肿瘤细胞内进行传播？ 3.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 努力开发新型生物标志物，并确定治疗靶标的关键蛋白质和过程。