Targeting HB-EGF and trigeminal EGFR for oral cancer pain and opioid tolerance

靶向 HB-EGF 和三叉神经 EGFR 治疗口腔癌疼痛和阿片类药物耐受

基本信息

批准号：
10582847
负责人：
Yi Ye
金额：
$ 238.97万
依托单位：
NEW YORK UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-19 至 2025-09-18
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10582847
关键词：
Absence of pain sensation Adjuvant Adverse effects Animals Arthritis Attenuated Behavior Biological Assay Bioluminescence Biosensor Brain Stem Cancer Model Cancer Patient Cell Line Cell model Cells Chemotherapy-Oncologic Procedure Chronic Clinical Clinical Trials Confocal Microscopy Coupling Cyclic AMP Receptors Data Development Disease Dose EGFR gene Effectiveness Electrophysiology (science)Endocytosis Endosomes Energy Transfer Epidermal Growth Factor Receptor Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor Erlotinib FDA approved Foundations Future GTP-Binding Proteins Gene Deletion Gene Expression Glutamates Gold Hyperactivity Image Immunofluorescence Immunologic Ligands Malignant Neoplasms Malignant neoplasm of pancreas Measures Mediating Mediator of activation protein Modeling Molecular Morphine Mus N-Methylaspartate N-methyl-D-glutamate Neurons Nociception Nociceptors Operative Surgical Procedures Opioid Oral Pain Pain Threshold Pain management Pathologic Patient Self-Report Patients Perioperative Peripheral Peripheral Nerves Pharmacology Phase Phase II Clinical Trials Placebos Preparation Prior Chemotherapy Proteins Quality of life Receptor Activation Receptor Protein-Tyrosine Kinases Receptor Signaling Reporter Resolution Role Sensory Signal Transduction Site Solid Neoplasm Spinal Stains Structure of trigeminal ganglion Synaptic Transmission System TRPV1 gene Testing Trigeminal System Tumor Tissue Western Blotting anti-cancer aspartate receptor base cancer pain cancer regression cancer therapy cancer type cell type chronic pain clinical translation cohort combat desensitization fluorescence imaging genetic approach heparin-binding EGF-like growth factor improved malignant mouth neoplasm motor behavior mouse model mu opioid receptors nanoparticle non-opioid analgesic novel opiate tolerance opioid use overexpression pain score presynaptic receptor receptor expression recruit single-cell RNA sequencing trafficking tumor

项目摘要

Project Summary/Abstract Oral cancer patients suffer from severe pain that is treated with opioids. However, opioid tolerance develops quickly. Currently, there are no other approaches to alleviate oral cancer pain or to forestall the development of opioid tolerance. In exploring a potential role of the epidermal growth factor receptor (EGFR) in oral cancer pain and opioid tolerance, we made an exciting discovery that one of the EGFR ligands, HB-EGF, is upregulated in patients with severe pain, and that EGFR interacts with both the mu opioid receptor (MOR) and the glutamate N-Methyl-D-aspartic acid receptor (NMDAR), two proteins important for chronic pain and opioid tolerance. This led us to hypothesize that HB-EGF mediated signaling in trigeminal (TG) neurons would: a) desensitize and down-regulate MOR and b) potentiate NMDAR-dependent synaptic transmission, contributing to oral cancer pain and opioid tolerance. We will test our hypothesis in three specific aims. Aim 1 will discover whether HB-EGF and trigeminal EGFR regulate pain, opioid tolerance, and NMDAR-mediated synaptic transmission in oral cancer. We will use pharmacological and genetic approaches to determine if HB-EGF and trigeminal EGFR are essential for oral cancer pain and opioid tolerance. Behaviors related to pain and opioid tolerance will be measured using a battery of assays. NMDAR mediated synaptic transmission will be studied using electrophysiological recordings in a newly developed brainstem preparation. We will quantify NMDAR expression in the mouse TG and brainstems using western blot. Aim 2 will discover whether HB-EGF and EGFR modulate MOR signaling and endocytosis. We will use BRET-based biosensors and super resolution imaging in model cell lines and TG neurons to study the impact of HB-EGF and EGFR on MOR cAMP activity, G-protein coupling, βARR recruitment and trafficking to endosomes. We will determine the effect of EGFR gene deletion and inhibition on MOR expression by qPCR, western blot, and fluorescence imaging in mouse TG neurons. Aim 3 will validate HB-EGF and EGFR as potential targets for pain and opioid tolerance in three oral cancer patient cohorts. First, we will model EGFR ligand expression in the tumor with self-reported pain, quantitative sensory testing scores, and opioid intake and explore the cellular origin of EGFR ligands by single cell RNA sequencing in tumor tissues. In the second cohort, we will model HB-EGF and EGFR expression in tumor tissues using immunofluorescence staining as a function of self-reported pain and opioid intake using data generated in a phase II anti-cancer trial examining the effect of an EGFR inhibitor erlotinib as an adjuvant to standard chemotherapy for cancer regression. In the third cohort, we will model peripheral EGFR and ligand(s) expression from self-reported pain. EGFR inhibitors are FDA-approved therapies for cancer management. HB-EGF is an emerging cancer target and a pain mediator. Targeting HB-EGF and EGFR therefore has the potential for rapid clinical translation to reduce oral cancer pain and opioid tolerance.

项目概要/摘要口腔癌患者在接受阿片类药物治疗时会遭受严重疼痛，但阿片类药物会产生耐受性。目前，没有其他方法可以减轻口腔癌疼痛或预防口腔癌的发展。阿片类药物耐受性，探索表皮生长因子受体 (EGFR) 在口腔癌中的潜在作用。疼痛和阿片类药物耐受性，我们做出了一个令人兴奋的发现，EGFR 配体之一 HB-EGF 是 EGFR 与 mu 阿片受体 (MOR) 和 mu 阿片受体 (MOR) 相互作用谷氨酸 N-甲基-D-天冬氨酸受体 (NMDAR)，两种对慢性疼痛和阿片类药物很重要的蛋白质这使我们得出结论，三叉神经元 (TG) 中 HB-EGF 介导的信号传导将：使 MOR 脱敏和下调，b) 增强 NMDAR 依赖性突触传递，有助于我们将在三个具体目标中检验我们的假设。 HB-EGF 和三叉神经 EGFR 是否调节疼痛、阿片类药物耐受性和 NMDAR 介导的突触我们将使用药理学和遗传学方法来确定 HB-EGF 是否与口腔癌中的传播有关。三叉神经 EGFR 对于口腔癌疼痛和阿片类药物耐受性至关重要。将使用一系列测定法来测量 NMDAR 介导的突触传递。我们将在新开发的脑干制剂中使用电生理记录来量化 NMDAR。使用蛋白质印迹目的 2 将发现 HB-EGF 和小鼠 TG 和脑干中的表达。 EGFR 调节 MOR 信号传导和内吞作用我们将使用基于 BRET 的生物传感器和超分辨率。在模型细胞系和 TG 神经元中进行成像，以研究 HB-EGF 和 EGFR 对 MOR cAMP 活性的影响， G 蛋白偶联、βARR 募集和转运至内体我们将确定 EGFR 的作用。 qPCR、蛋白质印迹和荧光成像对小鼠中基因缺失和 MOR 表达的抑制 TG 神经元将验证 HB-EGF 和 EGFR 作为三种疼痛和阿片类药物耐受性的潜在靶标。首先，我们将模拟具有自我报告疼痛的肿瘤中的 EGFR 配体表达，定量感官测试分数和阿片类药物摄入量，并通过单一方法探索 EGFR 配体的细胞起源在第二组中，我们将模拟肿瘤组织中的 HB-EGF 和 EGFR 表达。使用免疫荧光染色的肿瘤组织作为自我报告的疼痛和阿片类药物摄入量的函数 II 期抗癌试验中生成的数据，该试验检查 EGFR 抑制剂厄洛替尼作为佐剂的效果在第三组中，我们将模拟外周 EGFR 和癌症消退的标准化疗。 EGFR 抑制剂是 FDA 批准的癌症疗法。 HB-EGF 是一种新兴的癌症靶点和针对 HB-EGF 和 EGFR 的疼痛介质。因此具有快速临床转化以减少口腔癌疼痛和阿片类药物耐受性的潜力。