Investigation of the Wnt/PCP pathway in human malignant melanoma

应聘者描述(申请人提供)：应聘者为医学博士，2005年6月完成皮肤科临床培训，目前正在从事基础科学研究，目标是成为一名独立的内科科学家。这项研究是在华盛顿大学兰德尔·T·穆恩博士的赞助下进行的。月球实验室使用包括斑马鱼和青蛙在内的几个模型系统，研究Wnt信号通路在胚胎发育和人类疾病中的作用。拟议的培训将为候选人成为皮肤生物学和皮肤肿瘤学的独立研究员做好准备，重点是细胞生物学、细胞运动性和信号转导。恶性黑色素瘤的发病率比任何其他癌症增长得都快。转移性疾病缺乏有效的治疗方法，反映出我们对黑色素瘤的进展和转移的了解有限。最近的研究表明，Wnt信号通路是黑色素瘤进展和转移性疾病发展的重要调节因素。WNT是一种分泌蛋白，与细胞表面受体结合，触发一系列调控细胞内事件，调节细胞生长、细胞分化、细胞黏附和细胞运动等功能。这项建议的目的是利用黑色素瘤细胞株在Wnt表达后表现出更强的运动性，来确定在Wnt途径调节细胞运动中重要的蛋白质。该提案将利用已证明可行的技术来测试已知的Wnt蛋白对黑色素瘤细胞运动的影响。下一步，新的候选WNT蛋白将通过使用另一种成熟的技术纯化黑色素瘤细胞中的WNT相关蛋白复合体来识别。这些新的蛋白质也将被测试对细胞运动的影响。黑色素瘤进展中出现的分子异质性问题将通过评估在不同黑色素瘤细胞系背景下被证实具有功能意义的蛋白质来解决，并辅之以对患者黑色素瘤样本中的蛋白质表达的评估。总之，这些研究将有助于阐明恶性黑色素瘤背景下Wnt信号调节细胞运动的潜在机制，最终目标是发现可以作为预后生物标记物的蛋白质靶标和/或治疗操作的靶标。

DESCRIPTION (provided by applicant): The candidate is an MD-PhD who completed clinical training in dermatology in June 2005 and is now pursuing basic science research with the goal of becoming an independent physician-scientist. The research is being performed under the sponsorship of Dr. Randall T. Moon at the University of Washington. The Moon laboratory studies the role of the Wnt signaling pathway in embryonic development and human disease using several model systems, including zebrafish and frogs. The proposed training will prepare the candidate for a career as an independent investigator in skin biology and cutaneous oncology, with a focus on cell biology, cell motility and signal transduction. The incidence of malignant melanoma is increasing faster than that of any other cancer. The lack of effective therapies for metastatic disease reflects our limited understanding of the progression of melanoma and metastasis. Recent studies have implicated the Wnt signaling pathway as an important regulator of both melanoma progression as well as the development of metastatic disease. Wnt's are secreted proteins that bind cell surface receptors to trigger a series of regulated intracellular events that mediate functions including cell growth, cell differentiation, cell adhesion and cell motility. This proposal aims to identify proteins that are important in the regulation of cell motility by the Wnt pathway using a melanoma cell line that exhibits increased motility upon Wnt expression. The proposal will utilize techniques with demonstrated feasibility to test known Wnt proteins for their effects on melanoma cell motility. Next, new candidate Wnt proteins will be identified by the purification of Wnt-related protein complexes in melanoma cells using another established technique. These new proteins will also be tested for effects on cell motility. The issue of molecular heterogeneity seen in melanoma progression will be addressed by evaluating proteins with validated functional significance in the context of different melanoma cell lines, complemented by evaluation of protein expression in patient melanoma samples. Together, these studies will help elucidate the mechanisms underlying the regulation of cell motility by Wnt signaling in the context of malignant melanoma, with the ultimate goal of uncovering protein targets that could serve as biomarkers for prognosis and/or targets for therapeutic manipulation.