Epidermal keratinocytes mediate mechanical pain following neuropathic injury

表皮角质形成细胞介导神经性损伤后的机械性疼痛

• 批准号: 10387370
• 负责人: Alexander Mikesell
• 金额: $ 4.68万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10387370
• 关键词: Afferent Neurons; Agonist; Animals; Antineoplastic Agents; Behavioral; Behavioral Assay; Cells; Chemicals; Chemotherapy-induced peripheral neuropathy; Chronic; Data; Development; Electrophysiology (science); Epidermis; Exhibits; Fiber; Goals; Growth; Human; Hyperalgesia; Hypersensitivity; Injury; Ion Channel; Ion Channel Gating; Knockout Mice; Mass Spectrum Analysis; Mechanical Stimulation; Mechanics; Mediating; Molecular Biology; Mus; Nerve; Neuraxis; Neurons; Neuropathy; Paclitaxel; Pain; Patients; Peripheral; Peripheral Nervous System Diseases; Property; Role; Signal Transduction; Skin; Skin Tissue; Small Interfering RNA; Sodium; Tactile; Testing; Therapeutic; Tissue Donors; Touch sensation; Transgenic Mice; Work; afferent nerve; base; behavioral response; biomedical scientist; career; cell type; chronic neuropathic pain; chronic pain; inflammatory pain; injured; keratinocyte; knock-down; mechanical allodynia; mechanical stimulus; mechanotransduction; mouse model; nerve injury; painful neuropathy; patch clamp; pre-clinical; release factor; sensory neuropathy; side effect; skills; tissue injury; voltage

Project Summary Keratinocytes, the most abundant cell type in the epidermis, mediate normal touch sensation by detecting and encoding tactile information to sensory neurons. However, it is unknown if keratinocyte mechanotransduction contributes to chronic mechanical pain following tissue injury. My preliminary data suggest that keratinocytes isolated from nerve injured animals are sensitized to mechanical stimulation. Considering this finding, I propose to investigate if injury induced sensitization of keratinocyte mechanotransduction contributes to the development of touch evoked neuropathic pain. I will specifically examine if this sensitization is mediated by the mechanically gated ion channel PIEZO1, which my preliminary data suggests is a major keratinocyte mechanotransducer. Here in Aim 1, I hypothesize that sensitization of keratinocyte PIEZO1 contributes to the development of chronic mechanical pain in a mouse model chemotherapy-induced peripheral neuropathy (CIPN). I will utilize epidermal specific PIEZO1 knockout mice and mechanical behavioral assays to determine if loss of PIEZO1 protects against CIPN mechanical pain (Aim 1A). Additionally, I will utilize ex vivo skin nerve (Aim 1B) and whole-cell patch clamp (Aim 1C) recordings to determine if loss of keratinocyte PIEZO1 reduces CIPN sensory nerve and keratinocyte mechanical hypersensitivity. In Aim 2, I hypothesize that PIEZO1 mediates human keratinocyte mechanotransduction. I will utilize whole-cell patch electrophysiology and PIEZO1 targeted siRNA to determine if knockdown of PIEZO1 reduces human keratinocyte mechanical sensitivity (Aim 2A). Next, I will determine if incubation with the chemotherapeutic paclitaxel sensitizes human keratinocytes to mechanical stimulation and if PIEZO1 knockdown reduces this hypersensitivity (Aim 2B). Finally, I will use mass spectrometry to investigate signaling factors released from human keratinocytes by PIEZO1 activation under naïve conditions and following paclitaxel treatment (Aim 2C). Together these aims will determine if keratinocytes enhance evoked mechanical pain following neuropathic injury and if epidermal PIEZO1 may be potentially targeted to relieve neuropathic pain.

项目摘要 角质形成细胞，表皮中最丰富的细胞类型，通过检测和 将触觉信息编码为感觉神经元。但是，尚不清楚角质形成细胞机械转移是否存在 在组织损伤后导致慢性机械疼痛。我的初步数据表明角质形成细胞 从神经受伤的动物中分离出对机械刺激的敏感。考虑到这一发现，我提出了 研究损伤是否引起角质形成细胞机械转导的敏感性有助于发展 触摸引起的神经性疼痛。我将专门检查该灵敏度是否由机械介导 我的初步数据表明，门控离子通道压电1是主要的角质形成型机械转换器。 在AIM 1中，我假设角质形成细胞压电的灵敏度有助于慢性的发展 小鼠模型化疗引起的周围神经病（CIPN）中的机械疼痛。我会使用表皮 特定的压电敲除小鼠和机械行为分析，以确定压电1的损失是否保护 针对CIPN机械疼痛（AIM 1A）。此外，我将利用离体皮肤神经（AIM 1B）和全细胞 斑块夹（AIM 1C）记录以确定角质细胞压电的损失是否会减少CIPN感觉神经和 角质形成细胞机械性超敏反应。在AIM 2中，我假设Piezo1介导人角质形成细胞 机械转导。我将利用全细胞贴片电生理学和Piezo1靶向siRNA来确定 如果压电1的敲低会降低人角质形成细胞的机械灵敏度（AIM 2A）。接下来，我将确定是否 与化学治疗性紫杉醇一起孵育，将人角质形成细胞感知到机械刺激以及if 压电1敲低降低了这种超敏反应（AIM 2B）。最后，我将使用质谱法调查 在幼稚条件下通过压电激活从人角质形成细胞释放的信号传导因子 紫杉醇治疗（AIM 2C）。这些目标共同确定角质形成细胞是否增强了诱发的机械 神经性损伤后的疼痛，如果可能将表皮压电1靶向促进神经性疗法 疼痛。