Cell Biological Mechanisms of Melanoma Cell Motility in vivo

 描述（由申请人提供）：目前还不清楚黑素细胞如何从癌前痣转变为侵袭性肿瘤细胞;更少的研究是直接检查这种转变为体内运动性。斑马鱼允许肿瘤细胞运动性的高分辨率成像，并且斑马鱼具有与人类相似的黑素细胞分布。此外，癌前痣发展为早期黑色素瘤，后来发展为侵袭性肿瘤在鱼类和人类之间是保守的。因此，我将使用人在鱼异种移植方法以及转基因斑马鱼品系直接可视化黑色素瘤细胞的运动。我将重点关注肿瘤相关的巨噬细胞如何促进转移级联的特定步骤，特别是确定细胞生物学机制，利用斑马鱼的高分辨率成像的顺从性调节invadopodium形成。然后，我将验证我们的关键发现，在小鼠模型与成像窗口和命运映射与光转换技术。重要的是，我将发现炎症反应途径如何调节黑色素瘤转移，以更好地了解免疫细胞和黑色素瘤细胞如何在微环境中交流。该职业发展奖的候选人是一名博士后研究员，该资助申请中提出的研究将在Cecilia Moens博士的主要指导下进行，并在该奖项的K99阶段期间由John Condeelis博士（Albert Einstein College of Medicine，NY）共同指导。Fred哈钦森癌症研究中心的Moens实验室是进行这些研究的理想环境，因为它具有反向遗传学方法、细胞信号传导和斑马鱼胚胎活细胞成像策略的专业知识。Condeelis博士是肿瘤微环境领域的世界领导者，在肿瘤细胞侵袭的体外细胞骨架调控和小鼠模型中肿瘤细胞的活体成像方面都具有专业知识。我将从Moens和Condeelis博士的指导中受益匪浅，并将两位导师的专业知识用于理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一个世界知名的癌症研究所，并提供了丰富的癌症专业知识：我已经建立了合作与黑色素瘤专家和医生-科学家在弗雷德哈钦森癌症研究中心，在附近的华盛顿大学校园，并与黑色素瘤临床专家在MD安德森。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，特别是研究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Moens博士的实验室工作不到一年，K99奖将使我有足够的时间在她的实验室进一步开发这个模型，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肿瘤转移过程中调节黑色素瘤细胞运动的肿瘤间质信号传导机制。我的长期目标是发现黑色素瘤治疗的新标志物和治疗靶点。

 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.