Understanding how receptor tyrosine kinase activation dynamics specify proliferative cellular responses

了解受体酪氨酸激酶激活动力学如何指定增殖细胞反应

基本信息

批准号：
10678825
负责人：
Mark A Lemmon
金额：
$ 51.82万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-15 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10678825
关键词：
Address Affinity Agonist Biochemical Biological Assay Biophysics Breast Cancer cell line Cell Proliferation Cells Cessation of life Chimera organism Data Dimerization Endocytosis Epidermal Growth Factor Epidermal Growth Factor Receptor Epiregulin ErbB4 gene Feedback Future G-Protein-Coupled Receptors Goals Image Kinetics Ligands MAP Kinase Modules Malignant Neoplasms Mass Spectrum Analysis Measures Methods Microscopy Motivation Mutation Nature Nerve Growth Factors Neuregulins Oncogenic Outcome PC12 Cells Pathway interactions Phosphorylation Site Play Proliferating Property Receptor Activation Receptor Protein-Tyrosine Kinases Receptor Signaling Resistance Role Signal Induction Signal Pathway Signal Transduction Specific qualifier value Specificity T-Cell Receptor Testing Therapeutic Transducers Tyrosine Kinase Receptor Inhibition Work cancer cell design dimer epigen insight molecular imaging network models pharmacologic receptor response single molecule targeted treatment therapeutic target trafficking

项目摘要

Project Summary We propose to combine biophysical imaging and biochemical approaches to address two key unanswered questions in the field of signaling by receptor tyrosine kinases (RTKs) – a principal class of therapeutic targets in cancer where resistance to current therapies necessitates new pharmacological approaches: a. What defines the distinct set of responses, and cell fate, downstream of a particular RTK? b. How can different ligands for a given RTK promote distinct cellular responses? It is well known that epidermal growth factor (EGF) and nerve growth factor (NGF) promote proliferative and differentiative responses respectively in PC12 cells, through EGFR and TrkA, apparently using the same set of signaling pathways. Early studies showed that Erk activation kinetics plays a key role in determining the different cell fates induced by these ligands, with transient Erk activation being associated with proliferation and sustained Erk activation with differentiation. Rather than being defined solely by different feedback ‘wiring’ in the intracellular MAP kinase cascade, we and others have found that the RTK activation kinetics play a direct role in defining the dynamic properties of the signaling network. Several recent studies further argue that the strength of ligand-induced RTK dimers (and/or their lifetime) dictates signaling specificity, offering the possibility of dynamically-determined biased agonism in RTK signaling. To gain insight into the mechanistic basis for biased agonism in RTK signaling, we propose to elucidate how activating the same RTK intracellular region in different ways can result in dramatically different cellular responses (proliferation vs differentiation). Our previous structural, biophysical, and biochemical data suggest the hypothesis that signaling outcome is determined by RTK dimer stability and dynamics. Using intact and chimeric receptors, we will test this hypothesis by asking how altering RTK dimerization kinetics influences receptor endocytosis, post-endocytic trafficking, and dynamics of the downstream signaling network. We combine single-molecule imaging and microscopy studies of RTK trafficking with mass spectrometry and biochemical studies of downstream signaling networks to yield an integrated picture of this. Our primary motivation is to investigate how modifying RTK signaling dynamics (rather than simply inhibiting RTKs) might be used in future therapeutic approaches in cancer. Our Specific Aims address the following questions: 1 How can the same RTK intracellular region elicit orthogonal cellular responses depending on how it is activated? 2 What are the lifetimes of ErbB4/HER4 dimers induced by different neuregulin (NRG) ligands, and what is the basis for their sustained signaling? Our overall goal is to understand how dimerization dynamics can define signaling specificity, possibly through a kinetic proofreading similar to that seen in the T-cell receptor.

项目摘要我们建议将生物物理成像和生化方法结合起来，以解决两个未回答的关键受体酪氨酸激酶（RTK）的信号传导领域的问题 - 主要治疗靶标类在癌症中，对当前疗法的抵抗力必要的新药物方法：一个。是什么定义了特定RTK下游的不同响应集和细胞命运集？ b。给定RTK的不同配体如何促进不同的细胞反应？众所周知，表皮生长因子（EGF）和神经生长因子（NGF）促进了增殖和 PC12细胞中分别通过EGFR和TRKA分别使用相同集信号通路。早期研究表明，ERK激活动力学在确定这些配体诱导的不同细胞命运，瞬时ERK激活与增殖有关并通过分化持续ERK激活。而不是仅由不同的反馈“接线”来定义在细胞内地图激酶级联反应中，我们和其他人发现RTK激活动力学发挥了在定义信号网络的动态属性中的直接作用。最近的一些研究进一步认为配体引起的RTK二聚体的强度（和/或其寿命）决定了信号的特异性 RTK信号传导中动态确定的偏置激动剂的可能性。为了深入了解RTK信号中偏见激动剂的机理基础，我们建议阐明如何以不同的方式激活相同的RTK细胞内区域会导致巨大不同的细胞反应（增殖与分化）。我们以前的结构，生物物理和生化数据表明信号结果由RTK二聚体稳定性和动力学决定的假说。使用完整和嵌合受体，我们将通过询问RTK二聚动力学如何影响RTK二聚化来检验这一假设受体内吞作用，脑吞噬后运输和下游信号网络的动力学。我们将RTK运输的单分子成像和显微镜研究与质谱和下游信号网络的生化研究以产生综合图像。我们的主要动机是调查如何修改RTK信号传导动力学（而不是简单地抑制RTK）用于癌症的未来治疗方法。我们的具体目的解决以下问题： 1相同的RTK细胞内区域如何根据其的方式引起正交细胞反应活性？ 2不同的神经调节蛋白（NRG）配体引起的ERBB4/HER4二聚体的生命是什么？其持续信号的基础？我们的总体目标是了解如何通过类似于T细胞接收器中的动力学校对。