Activation and Signaling Mechanism of the RET Tyrosine Kinase Receptor

描述（由申请人提供）：该奖学金将支持波士顿大学阿德里安·惠蒂教授实验室的研究目标和研究生培训。许多形式的癌症是由生长因子（GF）受体的异常信号传导引起的。因此，实现对GF受体活化和功能的更好理解继续产生改进的癌症治疗方法。然而，关于GF受体活化和信号传导的分子机制的许多基本问题仍然没有答案。我们建议开发新的方法的定量机制的GF受体信号转导的研究，使用多组分的RE-排列在转染（RET）受体作为测试用例。RET是一种受体酪氨酸激酶，其是神经胶质细胞系衍生的神经营养因子（GDNF）家族的神经营养因子的受体。RET信号的失调与几种类型的癌症有因果关系，包括多发性内分泌瘤（MEN）2A和-2B、甲状腺髓样癌、甲状旁腺增生和嗜铬细胞瘤。我们提出通过追求以下三个目标来表征RET激活和信号传导的分子机制：1）基因编码的荧光共振能量转移（FRET）构建体将用于开发用于使用流式细胞术监测活细胞表面上的RET缔合事件的报告系统。2)在目标1中开发的FRET报告基因构建体将用于解决关于活细胞上RET激活机制的关键问题。这些问题包括解决困难和长期的不确定性，是否GF受体功能的配体诱导的受体二聚体，或者通过引起结构重排内预相关的受体二聚体或寡聚体。3)将探索RET和血管内皮生长因子受体2（VEGFR 2）之间串扰的机制和影响，以确定RET信号串扰发生在什么水平以及对RET功能的影响如何取决于VEGFR 2刺激水平。总的来说，这三个目标将建立新的方法来阐明GF受体激活的机制，并将建立两个不同的GF受体家族之间的串扰的机制和影响。这项研究将帮助研究员实现她的职业目标，即领导一个学术研究小组，解决与人类健康有关的重要生物学问题。她计划通过在分子，细胞和体内水平的系统复杂性，主要采用假设驱动和（在适当的情况下）定量的角度来解决这些问题。在她的研究生工作的培训计划将使她能够专注于发展她的知识和能力的分子和细胞的方法，与想法，她将继续通过在一个合适的实验室博士后研究涵盖体内技术和观点。候选人和她的担保人一起制定了一个非常有效的计划，以实现她实现长期职业目标所需的培训。

DESCRIPTION (provided by applicant): This fellowship will support the research aims and training of a graduate student in the laboratory of Professor Adrian Whitty at Boston University. Many forms of cancer are caused by aberrant signaling of growth factor (GF) receptors. Consequently, achieving an improved understanding of GF receptor activation and function continues to give rise to improved approaches to cancer treatment. However, many basic questions about the molecular mechanisms of GF receptor activation and signaling remain unanswered. We propose to develop new methods for the quantitative mechanistic study of GF receptor signaling, using the multicomponent RE-arranged during Transfection (RET) receptor as a test case. RET is a receptor tyrosine kinase that is the receptor for the glial cell-line deried neurotrophic factor (GDNF) family of neurotrophins. Deregulation of RET signaling is causally involved in several types of cancer, including multiple endocrine neoplasias (MEN) 2A and -2B, medullary thyroid carcinoma, parathyroid hyperplasia, and phaeochromocytoma. We propose to characterize the molecular mechanisms of RET activation and signaling by pursuing following three aims: 1) genetically encoded fluorescence resonance energy transfer (FRET) constructs will be used to develop a reporter system for monitoring RET association events on the surface of live cells using flow cytometry. 2) The FRET reporter constructs developed in Aim 1, will be used to address key questions concerning the mechanism of RET activation on live cells. These questions include addressing the difficult and longstanding uncertainty concerning whether GF receptors function by ligand-induced receptor dimerization, or alternatively by causing a structural rearrangement within pre-associated receptor dimers or oligomers. 3) The mechanisms and impact of crosstalk between RET and vascular endothelial growth factor receptor 2 (VEGFR2) will be explored, to establish at what levels in RET signaling crosstalk occurs and how the effects on RET function depend on the level of VEGFR2 stimulation. Collectively, these three aims will establish new methods to elucidate mechanisms of GF receptor activation, and will establish the mechanisms and impact of crosstalk between two different families of GF receptors. This research will help the fellow achieve her career goal, which is to lead an academic research group that addresses important biological problems relating to human health. She plans to address these problems by working at the molecular, cellular and in vivo levels of systems complexity, employing primarily a hypothesis-driven and (where appropriate) a quantitative perspective. The training plan in her graduate work will enable her to focus on developing her knowledge and capabilities of molecular and cellular approaches, with the idea that she will go on to encompass in vivo techniques and perspectives through post doctoral study in a suitable laboratory. Working together, the candidate and her sponsor have developed a highly effective plan for how to achieve the training she will need to meet her long-term career objectives.