Effects of GPCR trafficking on the spatiotemporal control of signaling

GPCR 转运对信号传导时空控制的影响

基本信息

批准号：
10330369
负责人：
Emily Elizabeth Blythe
金额：
$ 7.03万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-02-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10330369
关键词：
Address Affect Antineoplastic Agents Area Biological Biological Assay California Cancer Diagnostics Cell membrane Cell model Cell surface Cells Cellular biology Collaborations Communities Consequentialism Cues Cyclic AMP Cyclic AMP-Dependent Protein Kinases Development Drug Targeting Ductal Epithelial Cell Endocytosis FDA approved Family G Protein-Coupled Receptor Signaling G-Protein-Coupled Receptors GNAS gene GTP-Binding Protein alpha Subunits, Gs GTP-Binding Proteins Generations Genes Genetic Transcription Immune KRAS2 gene Lead Ligand Binding Ligands Location Logic Malignant Neoplasms Malignant neoplasm of pancreas Mass Spectrum Analysis Mentorship Molecular Mutate Mutation Neoplasm Metastasis Oncogenic Outcome Output Pancreatic Ductal Adenocarcinoma Pancreatic duct Pathway interactions Physiological Physiological Processes Physiology Post-Translational Protein Processing Process Proteins Proteomics Receptor Activation Receptor Signaling Research Role San Francisco Shapes Signal Transduction Site Stimulus System Testing Therapeutic Time Training Translational Research Tumor Biology Universities Variant Work base beta-2 Adrenergic Receptors cancer prevention cancer therapy insight neurotrophic factor new therapeutic target novel strategies pancreatic cancer cells pancreatic cancer model pancreatic ductal adenocarcinoma model receptor receptor-mediated signaling residence response spatiotemporal tool trafficking transcriptomics tumor

项目摘要

Project Summary/Abstract G protein coupled receptors (GPCRs) constitute a family of signaling receptors that regulate essentially every physiological process. Recent studies have begun to recognize the roles of G proteins and GPCRs in tumor biology. In particular, signaling by the β2-adrenergic receptor (β2-AR) and mutations in its cognate G protein, GNAS, have been associated with pancreatic ductal adenocarcinoma (PDAC) progression. Understanding the basic molecular mechanisms of GPCR signaling will be imperative for realizing their potential as novel therapeutic targets. Studies of ligand-dependent GPCR signaling have traditionally focused on receptor activation at the plasma membrane. However, the recent discovery that intracellular compartments also function as active signaling sites that elicit different signaling outcomes raises a critical biological question: how does GPCR trafficking dictate unique signals, and do these unique signals lead to unique cellular outcomes? The proposed work addresses this question by focusing on the role of endosomal GPCR activation in shaping cAMP- dependent transcriptional responses. First, starting in an established HEK293 cell model, the proposed studies will assess the mechanisms by which trafficking encodes the amount and timing of cAMP generation. Through manipulations of receptors and trafficking machinery, the relationship between endosomal residence time and cAMP signaling profiles will be tested to determine whether trafficking is sufficient to shape cAMP output or whether other factors, such as posttranslational modifications or interacting proteins, are required. Next, the proposed work will define how these distinct cAMP profiles are decoded to elicit a specific downstream response. Defined steps in the signaling cascade will be probed in order to ascertain whether the transcriptional response is sensitive to variations in cAMP generation or whether cells have evolved a uniform response to cAMP stimulus. Finally, the proposed studies will explore whether the principles delineated in the HEK293 system are relevant to our understanding of oncogenic signaling by β2-AR and GNAS in PDAC. The timing, location, and duration of cAMP generation in pancreatic ductal epithelial cells will be evaluated to determine if these cAMP signatures are regulated and consequential and how PDAC-associated mutations affect this relationship. These studies will advance an exciting area of fundamental cell biology—the spatiotemporal control of receptor-mediated signaling. The proposed research will also provide new insights into the mechanisms of oncogenic signaling by β2-AR and GNAS in PDAC and, more broadly, the paradigm of GPCR-induced cAMP signaling in cancer. This project will be carried out at the University of California, San Francisco under the mentorship of Dr. Mark von Zastrow, a leader in the field of GPCR trafficking and signaling, in collaboration with two experts in the fields of pancreatic cancer (Dr. Eric Collisson, UCSF) and oncogenic signaling by GPCRs (Dr. J. Silvio Gutkind, UCSD). In addition to the conceptual and experimental training from those labs, UCSF also offers a vibrant research community with expertise and facilities that will enable the successful realization of the proposed studies.

项目概要/摘要 G 蛋白偶联受体 (GPCR) 构成信号传导受体家族，本质上调节每一个信号通路最近的研究已经开始认识到 G 蛋白和 GPCR 在肿瘤中的作用。特别是 β2-肾上腺素受体 (β2-AR) 的信号传导及其同源 G 蛋白的突变， GNAS 与胰腺导管腺癌 (PDAC) 进展相关。 GPCR 信号传导的基本分子机制对于实现其作为新型药物的潜力至关重要配体依赖性 GPCR 信号传导的研究传统上集中于受体。然而，最近发现细胞内区室也发挥作用。由于引发不同信号结果的活性信号位点提出了一个关键的生物学问题：如何 GPCR 转运决定了独特的信号，这些独特的信号是否会导致独特的细胞结果？拟议的工作通过关注内体 GPCR 激活在塑造 cAMP-中的作用来解决这个问题首先，从已建立的 HEK293 细胞模型开始，所提出的研究。将评估贩运编码 cAMP 生成量和时间的机制。受体和运输机制的操纵，内体停留时间和将测试 cAMP 信号传导谱以确定运输是否足以塑造 cAMP 输出或是否需要其他因素，例如翻译后相互作用修饰或蛋白质。拟议的工作将定义如何解码这些不同的 cAMP 配置文件以引发特定的下游响应。将探测信号级联中的定义步骤，以确定转录反应是否对 cAMP 生成的变化或细胞是否进化出对 cAMP 刺激的统一反应敏感。最后，拟议的研究将探讨 HEK293 系统中描述的原则是否相关帮助我们了解 PDAC 中 β2-AR 和 GNAS 的致癌信号传导的时间、位置和持续时间。将评估胰腺导管上皮细胞中的 cAMP 生成，以确定这些 cAMP 特征是否存在这些研究将探讨 PDAC 相关突变如何影响这种关系。基础细胞生物学进展的一个令人兴奋的领域——受体介导的信号传导的时空控制。拟议的研究还将为 β2-AR 和 β2-AR 的致癌信号传导机制提供新的见解。 PDAC 中的 GNAS，以及更广泛的癌症中 GPCR 诱导的 cAMP 信号传导范例。该研究在加州大学旧金山分校马克·冯·扎斯特罗 (Mark von Zastrow) 博士的指导下进行。 GPCR 运输和信号传导领域的领导者，与胰腺领域的两位专家合作癌症（Eric Collisson 博士，UCSF）和 GPCR 的致癌信号传导（J. Silvio Gutkind 博士，UCSD）。除了这些实验室的概念和实验培训之外，加州大学旧金山分校还提供了一个充满活力的研究社区拥有能够成功实现拟议研究的专业知识和设施。