GPCR signaling during embryonic organ formation

胚胎器官形成过程中的 GPCR 信号传导

基本信息

批准号：
10584164
负责人：
Deborah J Andrew
金额：
$ 42.7万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2027-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10584164
关键词：
3-hydroxy-3-methylglutaryl-coenzyme A Actins Adult Affect Animals Apical Autonomic nervous system Cell Polarity Cell Shape Cell Survival Cell membrane Cell surface Cells Central Nervous System Collaborations Complement Conflict (Psychology)Cues Destinations Development Developmental Process Dictyostelium Disease Distant Drosophila garnet protein Drosophila genome Drosophila genus Drug Targeting Embryo Embryonic Development Enzymes Epithelium Erinaceidae Event Extracellular Matrix Extravasation F-Actin FDA approved Fogs G Protein-Coupled Receptor Signaling G-Protein-Coupled Receptors Genes Genetic Germ Cells Glutaminase Hormones Human Immune Immunity Inflammation Injury Life Ligands Light Link Mediating Membrane Molecular Molecular Mechanisms of Action Morphogenesis Neoplasm Metastasis Neurotransmitters Odors Organ Organism Organogenesis Oxidoreductase Pathway interactions Pattern Pharmaceutical Preparations Phenotype Pheromone Photons Play Polymers Positioning Attribute Process Proteins RNA Interference Receptor Activation Reporting Role Salivary Glands Signal Pathway Signal Transduction Site Smell Perception Smog Somatic Cell Source Stimulus Structure of primordial sex cell System Taste Perception Testing Time Tissues Tube Visual Wing Work cell motility constriction cost genome wide screen in vivo insight lipid phosphate phosphatase migration neuroblast novel polymerization pressure receptor receptor binding receptor function recruit response smoothened signaling pathway tissue/cell culture tool

项目摘要

G-protein coupled receptors (GPCRs) are critical for almost every aspect of animal life. These proteins are embedded in the cell membrane and allow us to sense and respond to light, smells, and taste. GPCRs also control responses in both our central and autonomic nervous systems, and they regulate both inflammation and immunity. GPCRs control cell migration for normal development and during cancer metastasis. Indeed, approximately 34% of FDA-approved drugs target GPCRs. Nonetheless, despite decades of study, many GPCRs have no known function, their ligands remain unidentified, and the pathways through which they elicit distinct cellular responses remain mostly uncharacterized. Here, we propose to take the first steps toward understanding the roles of GPCRs during development in the experimental system of the Drosophila embryo, which has numerous advantages in terms of visual accessibility, an extensive armamentarium of genetic tools, and relatively low cost. We begin with an analysis of the Drosophila GPCR Tre1, which has been implicated in germ cell (GC) navigation and survival, extravasation of immune cell to sites of injury, and polarization of neuroblasts. We have recently reported that non-canonical Hedgehog signaling works through the Tre1 receptor to control GC navigation, resolving a long-standing conflict regarding the role of Hh in this process and revealing a novel pathway downstream of Tre1 activation. In the first aim, we uncover the molecular and cellular mechanisms through which each step of this pathway is mediated – from receptor binding to actin polymerization. We ask if and how other genes that affect GC migration work through this pathway to repel GCs (in the case of the Wunen lipid phosphate phosphatases) or attract GCs (in the case of HMGCoA reductase). Tre1 is also expressed in the forming salivary gland (SG), a tissue that, unlike GCs, migrates as a fully polarized epithelial collective. We ask if Hh signaling and Tre1 also function in the SG for its navigation and we ask if Tre1 function in this tissue complements or antagonizes the function of another GPCR – Mthl5 – which is expressed in the SG at about the same time and that has also been implicated in Hh signaling. Finally, we establish a pipeline to screen all of the GPCRs encoded in the Drosophila genome and expressed in embryos for roles in the development of either GCs or the SG. Our pilot screen has already identified two GPCRs with phenotypes consistent with important functions, one gene with a potential role in GC survival and the other with a potential role in regulating the SG extracellular matrix.

G 蛋白偶联受体 (GPCR) 对于动物生命的几乎各个方面都至关重要。蛋白质嵌入细胞膜中，使我们能够感知光、气味、 GPCR 还控制我们的中枢神经系统和自主神经系统的反应，它们调节炎症和免疫，控制细胞的正常迁移。事实上，大约 34% 的 FDA 批准的药物。然而，尽管经过数十年的研究，许多 GPCR 仍没有已知的功能，它们的配体仍然未知，并且它们引发不同细胞的途径在此，我们建议采取初步措施。了解 GPCR 在实验系统发育过程中的作用果蝇胚胎在视觉可及性方面具有许多优势，遗传工具种类丰富，成本相对较低。我们首先分析果蝇 GPCR Tre1，它与细菌有关细胞 (GC) 导航和存活、免疫细胞外渗到损伤部位以及极化我们最近报道了非典型的 Hedgehog 信号传导。通过 Tre1 受体控制 GC 导航，解决了长期存在的冲突 Hh 在此过程中的作用并揭示了 Tre1 激活下游的新途径。第一个目标是，我们揭示每一步的分子和细胞机制途径是介导的——从受体结合到肌动蛋白聚合，我们询问是否以及如何其他。影响 GC 迁移的基因通过此途径排斥 GC（在 Wunen 脂质磷酸酶）或吸引 GC（在 HMGCoA 还原酶的情况下）。也在形成唾液腺 (SG) 中表达，该组织与 GC 不同，作为唾液腺迁移我们询问 Hh 信号传导和 Tre1 是否也在 SG 中发挥作用。导航，我们询问该组织中的 Tre1 功能是否补充或拮抗另一个 GPCR – Mthl5 – 大约在同一时间在 SG 中表达，并且也已最后，我们建立了一个筛选所有 GPCR 的管道。在果蝇基因组中编码并在胚胎中表达，在发育中发挥作用无论是 GC 还是 SG，我们的试点筛选已经鉴定出两个具有表型的 GPCR。与重要功能一致，一个基因在 GC 存活中具有潜在作用，而另一个基因具有调节 SG 细胞外基质的潜在作用。