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: 10670391
• 负责人: Shavonne Teng
• 金额: $ 4.85万
• 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/10670391
• 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; Clinical Treatment; Coupling; Dose; Drug Delivery Systems; Eating; Encapsulated; 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; Macrophage; Mechanics; Mediating; Mediator; Molecular; Mus; Neurons; Nociceptors; Opioid; Orofacial Pain; Oxidation-Reduction; PAR-2 Receptor; Pain; Pain management; Pathway interactions; Patients; Peptide Hydrolases; Pharmaceutical Preparations; Physiology; Pre-Clinical Model; Property; 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; antagonist; beta-arrestin; biophysical properties; calcium indicator; cancer pain; cancer therapy; career; chronic pain; chronic pain management; desensitization; design; experience; graduate student; malignant mouth neoplasm; mechanical allodynia; mechanical stimulus; 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，令人衰弱和慢性病，它破坏了患者的能力 吃和说话。患者通过连续使用阿片类药物来治疗癌症疼痛，并需要 升级剂量以实现缓解。阿片类药物还会产生严重的副作用，包括呼吸抑郁症， 成瘾和镇静。有必要采取替代性非阿片类药物策略。但是，口腔病理学 癌症疼痛目前尚不清楚。我的研究建议探讨了G蛋白的机制 偶联受体（GPCR）和瞬态受体电位（TRP）离子通道调节口腔癌疼痛。口服 癌细胞和免疫细胞，例如巨噬细胞的秘密蛋白酶，这些蛋白酶在细胞上清除并激活GPCR 表面。 GPCR激活和G蛋白耦合触发下游信号级联，可以感觉到 TRP通道。具体而言，GPCR蛋白酶激活的受体2（PAR2）介导了口腔癌的机械 异常性症和导致瞬时受体电势香草素4（TRPV4）激活和过度激活 伤害感受器。但是，来自质膜的GPCR信号传导是短暂的。 β逮捕蛋白脱敏 GPCR并将受体与网格蛋白介导的内吞作用，共同终止质膜 信号。传统上，内体被认为是受体回收和降解的渠道，但 我实验室的最新工作表明，内体中的GPCR继续产生持续的信号，表明 介导疼痛传播。但是，PAR2是否信号从内体信号到感知TRPV4并唤起 尚未确定过度兴奋性。这项研究的目的是确定 内体PAR2信号及其对口腔癌中TRPV4敏感性的影响。 pH-stimulus响应 将开发向疼痛感应三叉神经元内体传递PAR2拮抗剂的纳米颗粒 检查这一途径。 AIM 1将开发pH刺激性响应性纳米颗粒，并表征 带有PAR2拮抗剂的纳米颗粒的生物物理特性。 AIM 2将探讨 内体PAR2信号传导及其在TRPV4敏感性中的作用，通过遗传编码的生物传感器和钙 指标。 AIM 3试图评估口腔癌临床前小鼠模型中PAR2信号的机制。 通过pH-stimulus纳米颗粒研究内体PAR2信号传导将阐明内体GPCR是否是否 是口腔癌疼痛的有效治疗靶标。在完成本提案中概述的目标和培训计划时， 研究生Shavonne Teng将对与慢性有关的GPCR信号有深入的了解 疼痛。她将学习开发纳米医学，学习生物物理和细胞生物学的生物工程方法 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，并为她做好研究职业的准备 癌症疼痛场的独立研究者。