Osteoarthritis Progression and Sensory Pathway Alterations

骨关节炎进展和感觉通路改变

• 批准号: 9053984
• 负责人: RICHARD J MILLER
• 金额: $ 42.67万
• 依托单位: RUSH UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-01 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9053984
• 关键词: Afferent Neurons; Agonist; Behavior; C Fiber; CCL2 gene; Capsaicin; Characteristics; Chronic; Clinical; Complex; Degenerative polyarthritis; Disease; Disease Progression; Electrophysiology (science); Elements; Esthesia; Fiber; Goals; Health; Human; Hyperalgesia; Image; Imaging Techniques; Immunohistochemistry; Individual; Joints; Knee; Knee Osteoarthritis; Label; Location; Maintenance; Measurement; Mechanical Stimulation; Mechanics; Medial meniscus structure; Mediating; Medical; Modeling; Mus; Nerve Fibers; Neurons; Neuropathy; Nociception; Nociceptors; Opioid Receptor; Pain; Pathology; Pathway interactions; Patients; Pattern; Peripheral; Pharmacological Treatment; Physiological; Population; Property; Proprioception; Reporting; Secondary Hyperalgesias; Sensory; Signal Transduction; Spinal Ganglia; Staging; Stimulus; Symptoms; Techniques; Technology; Testing; Therapeutic; Time; Tissue Banking; Tissue Banks; Touch sensation; afferent nerve; behavior change; designer receptors exclusively activated by designer drugs; experience; in vivo; in vivo imaging; knee pain; mechanical allodynia; mouse model; novel; pain behavior; promoter; response; selective expression; sensory stimulus; somatosensory; spontaneous pain; vibration

DESCRIPTION (provided by applicant): Osteoarthritis (OA) pain represents an enormous health burden. Patients describe different types of pain over the course of the disease, including pain with mechanical characteristics, neuropathic elements, and intermittent spontaneous pain. Signs of somatosensory disturbances such as mechanical allodynia, loss of proprioception, and vibratory deficits have been reported. Specialized sensory neurons mediate different sensations, including touch, proprioception, vibration, and different types of noxious stimuli. Therefore, we propose that "pain" in OA is more than merely a state where nociceptive neurons in the joint are activated, but rather involves widespread changes in sensory neurons, both in specialized subsets of nociceptive neurons and in non-nociceptive A¿ fibers. Our overall goal is to identify these changes in subsets of neurons in the course of progressive OA. We will use a validated murine model, destabilization of the medial meniscus (DMM) that captures the long-term progression of OA. In this model, pain-dependent behaviors change over time, with knee hyperalgesia and mechanical allodynia presenting early on, while symptoms of ongoing pain are not apparent until 8-16 weeks post DMM. This provides a unique opportunity to longitudinally assess neuronal properties. Our strategy will include three different approaches: (1) To define changes in specific subsets of sensory neurons in dorsal root ganglia (DRG) and joints in the course of knee OA - Subsets of sensory neurons in the joint and innervating DRG, L2-L5, will be documented throughout the progression of disease, through retrograde labeling and immunohistochemistry. Specifically, we will describe the number and location of peptidergic and non-peptidergic C-fibers, of non-nociceptive A¿-fibers, and their expression of markers including CCR2 and Nav1.8. We will validate our findings in a unique human joint tissue bank including patients with meniscal pathology, early and advanced stages of knee OA; (2) To characterize physiological properties of subsets of sensory neurons during initiation and maintenance of OA pain- We will analyse activity of individual subsets of A and C-fibers through electrophysiological recordings on intact DRG, L3/L4, and determine spontaneous activity as well as responses to stimuli including MCP-1, capsaicin, and ATP. We will also employ novel Pirt-GCaMP3 mice to image intracellular Ca2+ in sensory neurons in vivo in response to natural sensory stimuli, including mechanical stimulation applied to the knee; (3) To study the effect of selectively silencing subsets of neurons on different OA pain behaviors - We will test the effect of a selective ¿-opioid receptor on pain-dependent behaviors. We will also employ novel DREADD technology to transiently silence all sensory DRG neurons (using the pan-DRG Advillin promoter) or a subset of nociceptors (using Nav1.8 promoter). The proposal combines neuroanatomical techniques, electrophysiological measurements, in vivo imaging, and state-of-the-art approaches to reversibly silence neurons in order to evaluate changes in populations of sensory neurons that contribute to symptoms in a translational OA model.

描述（由申请人提供）：骨关节炎（OA）疼痛代表着巨大的健康负担，患者描述了疾病过程中的不同类型的疼痛，包括具有机械特征的疼痛、神经性疼痛以及间歇性自发性疼痛的体征。据报道，机械性异常性疼痛、本体感觉丧失和振动缺陷会介导不同的感觉，包括触觉、本体感觉、振动和不同类型的感觉。因此，我们认为 OA 中的“疼痛”不仅仅是关节中伤害性神经元被激活的状态，而且还涉及感觉神经元的广泛变化，无论是在伤害性神经元的专门亚群中还是在非伤害性神经元中。 ¿我们的总体目标是确定进行性 OA 过程中神经元亚群的这些变化，我们将使用经过验证的小鼠模型，即内侧半月板 (DMM) 的不稳定来捕获 OA 的长期进展。 ，疼痛依赖性行为随着时间的推移而变化，膝盖痛觉过敏和机械异常性疼痛很早就出现，而持续疼痛的症状直到 DMM 后 8-16 周才明显，这提供了纵向评估的独特机会。我们的策略将包括三种不同的方法：(1) 定义膝关节 OA 过程中背根神经节 (DRG) 和关节中特定感觉神经元子集的变化 - 关节和支配 DRG 中的感觉神经元子集， L2-L5，将通过逆行标记和免疫组织化学记录整个疾病的进展，具体来说，我们将描述肽能和非肽能 C 纤维的数量和位置。非伤害性 A¿ -纤维及其标记物的表达，包括 CCR2 和 Nav1.8 我们将在一个独特的人类关节组织库中验证我们的发现，包括患有半月板病理、早期和晚期膝关节 OA 的患者 (2) 表征子集的生理特性； OA 疼痛发生和维持过程中感觉神经元的变化 - 我们将通过电生理学分析 A 纤维和 C 纤维各个子集的活动 我们还将使用新型 Pirt-GCaMP3 小鼠对体内感觉神经元的细胞内 Ca2+ 进行成像。自然感觉刺激，包括施加于膝盖的机械刺激；（3）研究选择性沉默神经元子集对不同 OA 疼痛行为的影响 - 我们将测试选择性沉默的效果-阿片受体对疼痛依赖性行为的影响。我们还将采用新颖的 DREADD 技术来暂时沉默所有感觉 DRG 神经元（使用泛 DRG Advillin 启动子）或伤害感受器的子集（使用 Nav1.8 启动子）。该提案结合了神经解剖学技术。 、电生理学测量、体内成像以及可逆地沉默神经元的最先进方法，以评估导致转化 OA 模型中症状的感觉神经元群体的变化。