Endosomal Signaling of PAR2 in Oral Cancer Pain

口腔癌疼痛中 PAR2 的内体信号转导

• 批准号: 10389741
• 负责人: Shavonne Teng
• 金额: $ 4.68万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2024-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10389741
• 关键词: Adopted; Biological Assay; Biomedical Engineering; Biophysics; Biosensor; Calcium; Cancer Etiology; Cancer Patient; Cell membrane; Cell surface; Cells; Cellular biology; Chemicals; Chronic; Clathrin; Cleaved cell; Clinical Treatment; Coupling; Dose; Drug Delivery Systems; Eating; Endocytosis; Endosomes; Fluorescence Resonance Energy Transfer; Future; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; GTP-Binding Proteins; Goals; Human; Immune; Impairment; Intractable Pain; Ion Channel; Learning; Ligands; Lipids; Mechanics; Mediating; Mediator of activation protein; Molecular; Mus; Neurons; Nociceptors; Opioid; Orofacial Pain; Oxidation-Reduction; PAR-2 Receptor; Pain; Pain management; Pathway interactions; Patients; Peptide Hydrolases; Pharmaceutical Preparations; Physiology; Plasma; Pre-Clinical Model; Receptor Activation; Recycling; Regulation; Reporting; Research; Research Personnel; Research Project Grants; Research Proposals; Research Training; Role; Sedation procedure; Signal Pathway; Signal Transduction; Signaling Protein; Spinal Ganglia; Stimulus; Structure of trigeminal ganglion; TRP channel; Therapeutic; Training; Trigeminal System; Vanilloid; Ventilatory Depression; Work; addiction; base; beta-arrestin; biophysical properties; calcium indicator; cancer pain; cancer therapy; career; chronic pain; desensitization; design; experience; graduate student; macrophage; malignant mouth neoplasm; mechanical allodynia; mouse model; nanomedicine; nanoparticle; nanoparticle delivery; non-opioid analgesic; orofacial; pain model; pain signal; pre-clinical; preclinical study; programs; receptor; receptor recycling; recruit; side effect; therapeutic evaluation; therapeutic target; transmission process

Oral cancer pain is a prevalent, debilitating, and chronic condition that disrupts patients’ ability to eat and speak. Patients develop tolerance from continuous use of opioids to treat cancer pain and require escalating doses to achieve relief. Opioids also produce severe side effects, including respiratory depression, addiction, and sedation. Alternative non-opioid strategies are necessary. However, the pathobiology of oral cancer pain is currently not well understood. My research proposal explores the mechanisms by which G-protein coupled receptors (GPCRs) and transient receptor potential (TRP) ion channels regulate oral cancer pain. Oral cancers and immune cells such as macrophages secrete proteases that cleave and activate GPCRs at the cell surface. GPCR activation and G protein coupling triggers downstream signaling cascades, which can sensitize TRP channels. Specifically, the GPCR protease-activated receptor-2 (PAR2) mediates oral cancer mechanical allodynia and leads to transient receptor potential vanilloid 4 (TRPV4) activation and hyperexcitability of nociceptors. However, GPCR signaling from the plasma membrane is transient. Beta arrestins desensitize GPCRs and couple the receptors for clathrin-mediated endocytosis, which together terminate plasma membrane signaling. While endosomes were traditionally thought of as a conduit for receptor recycling and degradation, recent work from my lab demonstrated that GPCRs in endosomes continue to generate sustained signals that mediate pain transmission. However, whether PAR2 signals from endosomes to sensitize TRPV4 and evoke hyperexcitability has not been established. The purpose of this research is to identify the mechanisms of endosomal PAR2 signaling and its effect on TRPV4 sensitization in oral cancer. pH-stimulus responsive nanoparticles that deliver PAR2 antagonists to endosomes of pain-sensing trigeminal neurons will be developed to examine this pathway. Aim 1 will develop pH-stimulus responsive nanoparticles and characterize the biophysical properties of the nanoparticles loaded with PAR2 antagonist. Aim 2 will explore the mechanisms of endosomal PAR2 signaling and its role in TRPV4 sensitization via genetically-encoded biosensors and calcium indicators. Aim 3 seeks to assess mechanisms of PAR2 signaling in pre-clinical mice models of oral cancer. Investigating endosomal PAR2 signaling via pH-stimulus nanoparticle will elucidate whether endosomal GPCRs are valid therapeutic targets for oral cancer pain. In completing the aims and training plan outlined in this proposal, the graduate student, Shavonne Teng, will gain a deep understanding of GPCR signaling relevant to chronic pain. She will learn bioengineering approaches to develop nanomedicines, learn biophysical and cell biology approaches to study GPCR and TRP regulation, and learn how to study orofacial pain in mouse models. This research training will expand on her past experience and also prepare her for a research career as an independent investigator in the cancer pain field.

口腔癌疼痛是一种普遍的、使人衰弱的慢性疾病，它破坏了患者 吃饭和说话。患者对持续使用阿片类药物治疗癌症疼痛产生耐受性， 逐步增加剂量以缓解症状阿片类药物也会产生严重的副作用，包括呼吸抑制， 成瘾和镇静替代性非阿片类药物策略是必要的。然而，口腔溃疡的病理生物学 癌症疼痛目前还没有得到很好的理解。我的研究计划是探索G蛋白 偶联受体（GPCR）和瞬时受体电位（TRP）离子通道调节口腔癌疼痛。口服 癌症和免疫细胞如巨噬细胞分泌蛋白酶， 面GPCR激活和G蛋白偶联触发下游信号级联，其可以致敏 TRP频道具体而言，GPCR蛋白酶激活受体-2（PAR 2）介导口腔癌的机械性 异常性疼痛，并导致瞬时受体电位香草酸4（TRPV 4）激活和过度兴奋， 伤害感受器然而，来自质膜的GPCR信号传导是瞬时的。β抑制蛋白脱敏 GPCR和偶联网格蛋白介导的内吞作用的受体，它们一起终止质膜 发信号。虽然内体传统上被认为是受体再循环和降解的管道， 我实验室最近的工作表明，核内体中的GPCR继续产生持续的信号， 介导疼痛传递。然而，PAR 2是否从内体发出信号来敏化TRPV 4并引起TRPV 4的激活， 尚未建立过度兴奋。本研究的目的是确定 内体PAR 2信号传导及其对口腔癌中TRPV 4致敏的作用。pH刺激响应 将开发一种纳米颗粒，将PAR 2拮抗剂递送到痛觉三叉神经元的内体 来研究这条路径目的1将开发pH刺激响应纳米颗粒并表征其性质。 图4示出了负载有PAR 2拮抗剂的纳米颗粒的生物物理性质。目标2将探讨 内体PAR 2信号传导及其在TRPV 4致敏中的作用，通过遗传编码的生物传感器和钙离子 指标目的3旨在评估口腔癌临床前小鼠模型中PAR 2信号传导的机制。 通过pH刺激纳米颗粒研究内体PAR 2信号传导将阐明内体GPCR是否 是口腔癌疼痛的有效治疗靶点。在完成本建议中概述的目标和培训计划时， 研究生Shavonne Teng将深入了解与慢性炎症相关的GPCR信号传导。 痛苦她将学习生物工程方法来开发纳米医学，学习生物物理学和细胞生物学 研究GPCR和TRP调节的方法，并学习如何在小鼠模型中研究口面疼痛。这 研究培训将扩大她过去的经验，也为她的研究生涯做好准备， 癌症疼痛领域的独立研究者