Protease/PAR2/TRPV4 Axis and Oral Cancer Pain

蛋白酶/PAR2/TRPV4轴与口腔癌疼痛

基本信息

批准号：
10321672
负责人：
NIGEL W BUNNETT
金额：
$ 70.73万
依托单位：
NEW YORK UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-01-01 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10321672
关键词：
Afferent Neurons Agonist Arrestins Attenuated Behavior Cancer Etiology Cancer Patient Caspase Cathepsins Cell Line Cell model Cell surface Cells Complex Coupling Development Disease Dose Eating Excision Exhibits Functional disorder G-Protein-Coupled Receptors GTP-Binding Proteins Human Impairment Intractable Pain Ion Channel Knowledge Lead Malignant Neoplasms Mechanics Mediating Mediator of activation protein Modeling Molecular Molecular Conformation Mus Neoplasm Metastasis Neurons Nociception Nociceptors Operative Surgical Procedures Opioid Oral PAR-2 Receptor Pain Pain management Pathway interactions Patients Peptide Hydrolases Pharmacology Play Protease Inhibitor Receptor Activation Receptor Signaling Recurrence Rodent Role Sedation procedure Sensory Serine Protease Signal Pathway Signal Transduction Site Specificity Speech Therapeutic Translations Trypsin Vanilloid Work afferent nerve antagonist asparaginylendopeptidase base cancer pain cell type design drinking extracellular human disease macrophage malignant mouth neoplasm mechanical allodynia mouse model neoplastic cell non-opioid analgesic novel opioid use orofacial pain score protease So receptor side effect spatiotemporal therapeutic target trafficking transmission process tumor tumor microenvironment

项目摘要

Oral cancer induces severe pain that disrupts talking, eating and drinking. Patients develop tolerance to the opioids used to treat this pain (mechanical allodynia); progressively larger doses are required. Unfortunately, opioids produce debilitating side effects including profound sedation. A non-opioid pharmacologic strategy to alleviate oral cancer pain is imperative; patients wait days or weeks before surgical resection, suffer from recurrence or cannot be cured. In previous work we found that protease-activated receptor-2 (PAR2) contributes to oral cancer mechanical allodynia. In more recent work we demonstrated that oral cancer and associated macrophages secrete proteases into the oral cancer microenvironment. We found that two poorly characterized proteases, legumain (Lgmn) and cathepsin-S (Cat-S), exhibit highly upregulated expression and activity in the oral cancer microenvironment and potentially mediate cancer pain. These proteases cleave PAR2 on sensory neurons. Cleavage at specific sites on PAR2 stabilizes receptor conformations which in turn promote activation of receptor signaling and trafficking pathways. These signaling pathways lead to TRPV4 activation and hyperexcitability of nociceptors. The work we now propose will define mechanisms of Lgmn and Cat-S in oral cancer pain. We propose to identify and localize activated proteases in oral cancers using our fluorescently-quenched activity-based probes (qABPs) that covalently interact with activated proteases. We will unequivocally identify and determine the cellular origin of activated proteases present in tumors from patients with oral cancer. We will then analyze the correlation between protease activity and pain scores, and determine whether tumor proteases induce PAR2-dependent activation of nociceptors. Using mice that lack PAR2 or TRPV4 in Nav1.8 nociceptors, and our validated models of oral cancer pain, we will determine whether tumor cell and macrophage proteases cause pain by activating PAR2 and TRPV4 on nociceptors. Furthermore, we will use our new Par2-muGFP mice for PAR2 localization and trafficking with high specificity and spatiotemporal fidelity. Lastly we will define the mechanisms by which tumor cell and macrophage proteases activate PAR2 and TRPV4, induce hyperexcitability of nociceptors, and cause oral nociception. We assert that protease inhibitors as well as PAR2 and TRPV4 antagonists hold therapeutic potential for oral cancer pain; we ultimately seek to exploit these mechanisms to develop cancer pain therapies.

口腔癌会引起剧烈疼痛，影响说话、饮食。患者对药物产生耐受性用于治疗这种疼痛的阿片类药物（机械性异常性疼痛）；需要逐渐加大剂量。很遗憾，阿片类药物会产生使人衰弱的副作用，包括深度镇静作用。非阿片类药物的药理学策略缓解口腔癌疼痛势在必行；患者在手术切除前等待数天或数周，遭受复发或无法治愈。在之前的工作中我们发现蛋白酶激活受体2（PAR2）导致口腔癌机械性异常性疼痛。在最近的工作中，我们证明口腔癌和相关的巨噬细胞将蛋白酶分泌到口腔癌微环境中。我们发现有两个不好特征蛋白酶，legumain (Lgmn) 和组织蛋白酶-S (Cat-S)，表现出高度上调的表达和口腔癌微环境中的活性并可能介导癌症疼痛。这些蛋白酶会裂解感觉神经元上的 PAR2。 PAR2 上特定位点的切割稳定了受体构象，从而稳定了受体构象促进受体信号传导和运输途径的激活。这些信号通路导致TRPV4 伤害感受器的激活和过度兴奋。我们现在提出的工作将定义 Lgmn 和 Cat-S 治疗口腔癌疼痛。我们建议使用我们的技术来识别和定位口腔癌中的活化蛋白酶与激活的蛋白酶共价相互作用的荧光淬灭活性探针（qABP）。我们将明确识别和确定患者肿瘤中存在的活化蛋白酶的细胞起源患有口腔癌。然后我们将分析蛋白酶活性和疼痛评分之间的相关性，以及确定肿瘤蛋白酶是否诱导 PAR2 依赖性伤害感受器激活。使用缺少的小鼠 Nav1.8 伤害感受器中的 PAR2 或 TRPV4，以及我们经过验证的口腔癌疼痛模型，我们将确定肿瘤细胞和巨噬细胞蛋白酶是否通过激活伤害感受器上的 PAR2 和 TRPV4 引起疼痛。此外，我们将使用新的 Par2-muGFP 小鼠进行 PAR2 定位和高特异性运输和时空保真度。最后我们将定义肿瘤细胞和巨噬细胞的机制蛋白酶激活 PAR2 和 TRPV4，诱导伤害感受器过度兴奋，并引起口腔伤害感受。我们断言蛋白酶抑制剂以及 PAR2 和 TRPV4 拮抗剂具有口服治疗的潜力癌症疼痛；我们最终寻求利用这些机制来开发癌症疼痛疗法。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Oral cancer patients experience mechanical and chemical sensitivity at the site of the cancer.

DOI：
10.1186/s12885-022-10282-3
发表时间：
2022-11-11
期刊：
BMC cancer
影响因子：
3.8
作者：
通讯作者：

Application of a Sulfoxonium Ylide Electrophile to Generate Cathepsin X-Selective Activity-Based Probes.

DOI：
10.1021/acschembio.9b00961
发表时间：
2020-03-20
期刊：
ACS chemical biology
影响因子：
4
作者：
Mountford SJ;Anderson BM;Xu B;Tay ESV;Szabo M;Hoang ML;Diao J;Aurelio L;Campden RI;Lindström E;Sloan EK;Yates RM;Bunnett NW;Thompson PE;Edgington-Mitchell LE
通讯作者：
Edgington-Mitchell LE

A disintegrin and metalloproteinase domain 17-epidermal growth factor receptor signaling contributes to oral cancer pain.