Cell Biological Mechanisms of Melanoma Cell Motility in vivo

 描述(申请人提供)：尚不清楚黑素细胞如何从癌前痣转变为侵袭性肿瘤细胞；更少的研究是在体内直接检查这种转变为运动性的。斑马鱼可以对肿瘤细胞的运动进行高分辨率成像，而斑马鱼的黑素细胞分布与人类相似。此外，从癌前痣到早期黑色素瘤，再到侵袭性肿瘤的进展在鱼类和人类之间是保守的。因此，我将使用人在鱼中异种移植的方法以及转基因斑马鱼品系来直接观察黑色素瘤细胞的运动。我将专注于肿瘤相关巨噬细胞如何促进转移级联反应的特定步骤，特别是通过利用斑马鱼高分辨率成像的适应性来识别调节Inposodium形成的细胞生物学机制。然后，我将用成像窗口验证我们在小鼠模型中的关键发现，并用光转化技术验证命运映射。重要的是，我将发现炎症反应通路如何调节黑色素瘤的转移，以更好地了解免疫细胞和黑色素瘤细胞如何在微环境中进行沟通。这项职业发展奖的候选人是一名博士后，这项资助申请中提出的研究将在Cecilia Moens博士的主要指导下进行，并在K99阶段的奖励期间在John Condeelis博士(纽约阿尔伯特·爱因斯坦医学院)的共同指导下进行。位于弗雷德·哈钦森癌症研究中心的莫恩斯实验室拥有斑马鱼胚胎反向遗传学方法、细胞信号和活细胞成像策略方面的专业知识，是进行这些研究的理想环境。Condeelis博士是肿瘤微环境领域的世界领先者，在肿瘤细胞侵袭的体外细胞骨架调控和小鼠模型肿瘤细胞的活体成像方面拥有专业知识。我将从Moens博士和Condeelis博士的指导中受益匪浅，并将把两位导师的专业知识联系起来，以了解鱼类和小鼠模型中调节黑色素瘤进展的动态细胞-细胞相互作用。弗雷德·哈钦森癌症研究中心是世界著名的癌症研究所，提供丰富的癌症专业知识：我与弗雷德·哈钦森癌症研究中心附近华盛顿大学校园的黑色素瘤专家和内科科学家以及MD Andersen的黑色素瘤临床专家建立了合作关系。我致力于在一家研究癌细胞生物学的学术机构担任独立研究员的职业，特别是研究在转移过程中调节肿瘤细胞运动的动态细胞骨架机制。我目前的博士后奖学金将于2014年8月结束。由于我在莫恩斯博士的实验室只工作了不到一年，K99奖将给我足够的时间在她的实验室进一步开发这个模型，让我为我的独立实验室正在进行的项目奠定基础。我的近期目标是确定在转移过程中调节黑色素瘤细胞运动的肿瘤间质信号机制。我的长期目标是发现治疗黑色素瘤的新标记物和治疗靶点。

 DESCRIPTION (provided by applicant): It is unclear how a melanocyte transitions from a premalignant nevus to an invasive tumor cell; even less studied is the direct examination of this transition to motility in vivo. Zebrafish allows for high resolution imaging of tumor cell motility and zebrafish have melanocyte distribution that is similar to humans. Additionally, the progression of premalignant nevi to an early melanoma and later to an aggressive tumor is conserved between fish and humans. Thus, I will use human-in-fish xenotransplantation approaches as well as transgenic zebrafish strains to directly visualize melanoma cell motility. I will focus on how tumor associated macrophages facilitate specific steps of the metastatic cascade, specifically identifying the cell biological mechanisms regulating invadopodium formation by taking advantage of the amenability to high resolution imaging of zebrafish. I will then validate our key findings in mouse models with imaging window and fate mapping with photoconversion techniques. Importantly, I will discover how inflammatory response pathways regulate melanoma metastasis to better understand how immune cells and melanoma cells communicate in the microenvironment. The candidate for this career development award is a postdoctoral fellow, and the research proposed in this grant application will be conducted under the primary mentorship of Dr. Cecilia Moens, and under the co- mentorship of Dr. John Condeelis (Albert Einstein College of Medicine, NY) during the K99 phase of the award. The Moens lab at the Fred Hutchinson Cancer Research Center is an ideal environment for these studies due to the expertise of reverse genetics approaches, cell signaling, and live cell imaging strategies in zebrafish embryos. Dr. Condeelis is a world leader in the tumor microenvironment, and has expertise in both in vitro cytoskeletal regulation of tumor cell invasion, and intravital imaging of tumor cells in murine models. I will benefit immensely from the mentorship of Drs. Moens and Condeelis, and will bridge the expertise of both mentors toward understanding the dynamic cell-cell interactions regulating melanoma progression in fish and mouse models. The Fred Hutchinson Cancer Research Center is a world renowned cancer institute, and provides a wealth of cancer expertise: I have established collaborations with melanoma experts and physician- scientists within the Fred Hutchinson Cancer Research Center, at nearby University of Washington campuses, and with a melanoma clinical expert at MD Andersen. I am committed to a career as an independent investigator at an academic institution studying cancer cell biology, specifically studying the dynamic cytoskeletal mechanisms regulating tumor cell movements during metastasis. My current postdoctoral fellowship ends in August 2014. As I only been in Dr. Moens' lab for less than a year, the K99 award will give me enough time in her lab to develop this model further for me to set up the foundation for ongoing projects in my independent lab. My immediate goal is to define the tumor stromal signaling mechanisms regulating melanoma cell motility during metastasis. My long-term goal is to discover new markers and therapeutic targets for melanoma treatment.