Endosomal mechanisms signaling oral cancer pain

口腔癌疼痛的内体机制

• 批准号: 10786660
• 负责人: NIGEL W BUNNETT
• 金额: $ 482.06万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-19 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10786660
• 关键词: Acidity; Address; Agonist; Biological Assay; Bioluminescence; Biophysics; Biopsy; Biosensor; Cancer Patient; Cell Surface Receptors; Cell membrane; Cell surface; Cells; Chemicals; Chronic Disease; Clathrin; Clinical Trials; Complex; Consent; Disease; Eating; Encapsulated; Endocytosis; Endosomes; Energy Transfer; Engineering; Environment; Epidermal Growth Factor Receptor; Excision; Failure; G-Protein-Coupled Receptors; Growth Factor; Human; Immune; Ion Channel; Ligands; Light; Link; Location; Malignant Neoplasms; Mechanics; Mediating; Membrane; Monoclonal Antibodies; Mus; Nerve; Neurobiology; Neurons; Neuropeptides; Nociception; Nociceptors; Operative Surgical Procedures; Organ Donor; Pain; Pathologic Processes; Patients; Peptide Hydrolases; Phenotype; Physiological Processes; Pre-Clinical Model; Questionnaires; Receptor Protein-Tyrosine Kinases; Resected; Rodent Model; Scaffolding Protein; Schwann Cells; Signal Pathway; Signal Transduction; Signaling Protein; Stimulus; Symptoms; Testing; Therapeutic; Trigeminal Nerve Neoplasm; antagonist; cancer pain; chronic pain; chronic pain management; comorbidity; confocal imaging; drinking; drug discovery; effective therapy; extracellular; improved; individual patient; inhibitor; insight; lingual nerve; malignant mouth neoplasm; mechanical allodynia; nanoparticle; pain model; pain signal; pre-clinical; preclinical study; receptor; response; side effect; small molecule; targeted treatment; therapeutic target; translational approach; tumor; tumor microenvironment

PROJECT SUMMARY/ABSTRACT GPCRs and RTKs are transmembrane receptors that control a host of physiological and pathological processes including cancer pain and other forms of chronic pain. This proposal focuses on signaling mechanisms of GPCRs and RTKs located in endosomes (eGPCRs and eRTKs) that might be responsible for cancer pain. The underlying hypothesis is that eGPCRs and eRTKs generate long-lasting signals in endosomes of human nociceptors and Schwann cells that induce sustained activation and sensitization to mediate pain. Cells within the oral cancer microenvironment (cancer, immune, and neuronal cells), release proteases, neuropeptides and growth factors that activate GPCRs (e.g., PAR2, CLR) and RTKs (e.g., TrkA, EGFR) on nociceptors and Schwann cells. GPCRs and RTKs undergo clathrin-dependent endocytosis and assemble signaling complexes that control activity and expression of ion channels and long-lasting sensitization of nociceptors and Schwann cells to drive pain. A reverse translational approach is used to study endosomal signaling of pain in 3 Specific Aims. Aim 1 determines the contribution of eGPCR and eRTK signaling to pain. Oral cancer patient pain will be evaluated (questionnaires, sensitivity testing of patient oral cancer tumors with mechanical and chemical stimuli, and assessment of comorbidities). Surgically resected tumors, trigeminal nerve, and cancer supernatant will be collected. Patient-specific Schwann cells will be isolated and cultured from the nerve. Nociceptors and Schwann cells will be obtained from organ donors. Nociceptive effects of cancer supernatant, proteases, neuropeptides, and growth factors that mediate activity of nociceptors and Schwann cells will be studied in mice. Endocytosis inhibitors will be used to probe the link between endocytosis and nociception. Sensitization and nociception will be correlated with individual patient pain phenotypes. Aim 2 Defines eGPCR and eRTK pain signaling mechanisms. Effects of cancer supernatant, and eGPCR and eRTK ligands will be studied. Endocytosis of GPCRs and RTKs that mediate oral cancer pain will be studied in human and mouse nociceptors and Schwann cells by confocal imaging and biophysical assays. Biosensors will be used to study the assembly of multiprotein signaling complexes in subcellular compartments of nociceptors and Schwann cells. Endosomal signaling will be correlated with pain phenotypes in individual patients. Aim 3 develops nanoparticles (NP) to antagonize eGPCR and eRTK signaling in preclinical pain models that recapitulate patient oral cancer pain. Endosomal acidity and light will be exploited to trigger NP disassembly and antagonist release, and test contributions of eGPCR and eRTK signaling. Their use in preclinical pain models will link endosomal signaling to pain. A deeper understanding of eGPCR and eRTK signaling mechanisms will provide insight into the neurobiology of oral cancer pain.

项目概要/摘要 GPCR 和 RTK 是控制一系列生理和病理的跨膜受体 过程包括癌症疼痛和其他形式的慢性疼痛。该提案的重点是信令 位于内体中的 GPCR 和 RTK 的机制（eGPCR 和 eRTK）可能是造成这种情况的原因 用于癌症疼痛。基本假设是 eGPCR 和 eRTK 在以下环境中产生持久信号： 人类伤害感受器和雪旺细胞的内体诱导持续激活和敏化 调解疼痛。口腔癌微环境中的细胞（癌症细胞、免疫细胞和神经元细胞）释放 激活 GPCR（例如 PAR2、CLR）和 RTK（例如 TrkA、 EGFR）对伤害感受器和雪旺细胞的影响。 GPCR 和 RTK 经历网格蛋白依赖性内吞作用 组装控制离子通道活性和表达的信号复合物，并持久 伤害感受器和雪旺细胞的敏化以驱动疼痛。逆向翻译方法用于 研究 3 个具体目标中疼痛的内体信号传导。目标 1 确定 eGPCR 的贡献和 eRTK 向疼痛发出信号。将评估口腔癌患者的疼痛（调查问卷、敏感性测试） 患者口腔癌肿瘤的机械和化学刺激，以及合并症的评估）。 将收集手术切除的肿瘤、三叉神经和癌症上清液。患者特定 将从神经中分离并培养雪旺细胞。将获得伤害感受器和雪旺细胞 来自器官捐献者。癌症上清液、蛋白酶、神经肽和生长的伤害性影响 将在小鼠中研究介导伤害感受器和雪旺细胞活性的因素。内吞抑制剂 将用于探讨内吞作用和伤害感受之间的联系。敏化和伤害感受将会 与个体患者的疼痛表型相关。目标 2 定义 eGPCR 和 eRTK 疼痛信号传导 机制。将研究癌症上清液以及 eGPCR 和 eRTK 配体的影响。内吞作用 将在人类和小鼠伤害感受器中研究介导口腔癌疼痛的 GPCR 和 RTK 通过共聚焦成像和生物物理测定来检测雪旺细胞。生物传感器将用于研究组装 伤害感受器和雪旺细胞亚细胞区室中多蛋白信号传导复合物的研究。 内体信号传导将与个体患者的疼痛表型相关。目标 3 制定 纳米粒子 (NP) 在临床前疼痛模型中拮抗 eGPCR 和 eRTK 信号传导 患者口腔癌疼痛。内体酸度和光将被用来触发 NP 分解和 拮抗剂释放，并测试 eGPCR 和 eRTK 信号传导的贡献。它们在临床前疼痛中的应用 模型将内体信号传导与疼痛联系起来。更深入地了解 eGPCR 和 eRTK 信号传导 机制将有助于深入了解口腔癌疼痛的神经生物学。