CaMKII as a Mechanism & Intervention Target for Sickle Cell Pain

CaMKII 作为一种机制

• 批准号: 7765396
• 负责人: Zaijie Jim Wang
• 金额: $ 44.82万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7765396
• 关键词: Acute; Adverse effects; African; Age; Animals; Behavior; Biological Markers; Biometry; Birth; Chemicals; Chemistry; Chronic; Clinical; Data; Development; Disease; Effectiveness; Formalin; Freund' s Adjuvant; Genetic; Goals; Heating; Human; Immunoblotting; Inflammatory; Informatics; Intervention; Kinetics; Knock-out; Knockout Mice; Knowledge; Lead; Ligation; Literature; Mechanics; Methods; Modeling; Molecular; Mus; Mutant Strains Mice; Mutation; Neurobiology; Neurons; Outpatients; Pain; Pain Measurement; Paper; Pathway interactions; Patient Self-Report; Patients; Persistent pain; Pharmaceutical Preparations; Pharmacological Treatment; Pharmacology; Pharmacy facility; Phenotype; Pilot Projects; Probability; Publishing; Reporting; Research Personnel; Safety; Sampling; Sensory; Sickle Cell; Sickle Cell Anemia; Small Interfering RNA; Spinal; Spinal nerve structure; Stimulus; Tactile; Tail; Testing; Thermal Hyperalgesias; Time; Toxicogenetics; Transgenic Mice; Transgenic Organisms; Translations; abstracting; age related; allodynia; anesthesiology nursing; base; biobehavior; calmodulin-dependent protein kinase II; chronic pain; computerized; effective therapy; hemoglobin A(0); inflammatory neuropathic pain; inhibitor/antagonist; innovation; interdisciplinary approach; knockout gene; mouse model; neuromechanism; pain behavior; prevent; primary outcome; response; secondary outcome; sex; sickling; spontaneous pain; success; tool; translational study

DESCRIPTION (provided by applicant): Our long-term goal is to advance knowledge of the neural mechanisms of pain in sickle cell disease (SCD) and develop an effective pharmacologic treatment. The neurobiology of pain in SCD is poorly understood. Several transgenic/knock-out mouse models have been successfully produced and applied to study SCD. However, pain findings were only recently published that indicated increased pain sensitivity in NY1DD mice using a radiant heat tail-flick test. Strikingly, pain in these mice was age-dependent with an onset time around 6 weeks. Unfortunately, the investigators did not report other pain measurement findings or define a pain phenotype in Berkeley sickle transgenic mice, a model of a more severe form of SCD. In our own preliminary studies, we examined Berkeley sickle mice and littermate non-sickle controls in an array of pain tests that are used to study other pain types in animals. In these preliminary studies, we found the presence of tactile allodynia and thermal hyperalgesia and that spinal CaMKII expression and activity were upregulated, similar to what we observed in other mouse models of inflammatory and neuropathic pain. In the latter models, we have identified CaMKIIa to be a critical component leading to persistent pain. We observed that spinal nerve ligation-induced pain behaviors did not develop in CaMKIIa mutant mice. Based on these encouraging data, we propose to extensively characterize pain behaviors in Berkeley mice by employing standard pain tests for spontaneous pain behaviors and those evoked by thermal or mechanical stimuli (von Frey, Hargreaves, hot-plate, tail-flick, cold allodynia) and inflammatory stimuli (formalin, complete Freund's adjuvant) from shortly after birth through adulthood or when responses plateau. Some of these pain tests are used in our ongoing human studies of SC pain using quantitative sensory testing (QST). Intentionally, these similarities will allow us to interpret findings from the SCD transgenic mouse model with consideration of findings from other mouse models of pain and human SCD pain. We will use real time PCR, immunoblotting, immunohisto-chemistry, and enzymatic kinetics methods to systematically examine the expression and activity of CaMKIIa and total CaMKII in sickle and control mice and correlate the changes in this potential biomarker with the onset of pain. To directly test the hypothesis that spinal CaMKIIa is a molecular mechanism that promotes and maintains the manifestation of chronic pain in SCD, we will conduct pharmacological studies to inhibit CaMKIIa using chemical, small interfering RNA (siRNA), and gene knockout methods. We propose to test in these pharmacological studies a clinically used orally available drug that we have found to be a CaMKII inhibitor and to reduce pain behaviors in inflammatory and neuropathic pain models. Our secondary strategy is to conduct a pilot translational study to identify safety issues and clinical potential of this CaMKII inhibitor by characterizing sensory pain in humans with quantitative sensory testing and a computerized self-report tool. The significance of this proposal is that it may ultimately lead to pharmacological interventions that target the CaMKII-pathway. (End of Abstract)

描述（由申请人提供）： 我们的长期目标是增进对镰状细胞病 (SCD) 疼痛神经机制的了解并开发有效的药物治疗方法。对 SCD 疼痛的神经生物学知之甚少。几种转基因/基因敲除小鼠模型已成功制备并应用于研究 SCD。然而，疼痛研究结果最近才发表，表明使用辐射热甩尾测试，NY1DD 小鼠的疼痛敏感性增加。引人注目的是，这些小鼠的疼痛与年龄有关，发作时间约为 6 周。不幸的是，研究人员没有报告其他疼痛测量结果或定义伯克利镰状转基因小鼠（一种更严重的 SCD 形式的模型）的疼痛表型。在我们自己的初步研究中，我们在一系列用于研究动物其他疼痛类型的疼痛测试中检查了伯克利镰状小鼠和同窝非镰状小鼠。在这些初步研究中，我们发现存在触觉异常性疼痛和热痛觉过敏，并且脊髓 CaMKII 表达和活性上调，类似于我们在其他炎症和神经性疼痛小鼠模型中观察到的情况。在后面的模型中，我们已经确定 CaMKIIa 是导致持续性疼痛的关键成分。我们观察到，CaMKIIa 突变小鼠没有出现脊神经结扎引起的疼痛行为。基于这些令人鼓舞的数据，我们建议通过采用标准疼痛测试来广泛表征伯克利小鼠的疼痛行为，以检测自发性疼痛行为以及从出生后不久到成年或反应时由热或机械刺激（冯弗雷、哈格里夫斯、热板、甩尾、冷异常性疼痛）和炎症刺激（福尔马林、完全弗氏佐剂）引起的疼痛行为 高原。其中一些疼痛测试用于我们正在进行的使用定量感觉测试 (QST) 的 SC 疼痛人体研究。有意地，这些相似之处将使我们能够解释 SCD 转基因小鼠模型的发现，同时考虑其他小鼠疼痛模型和人类 SCD 疼痛的发现。我们将使用实时 PCR、免疫印迹、免疫组织化学和酶动力学方法来系统地检查镰状小鼠和对照小鼠中 CaMKIIa 和总 CaMKII 的表达和活性，并将这种潜在生物标志物的变化与疼痛的发作相关联。为了直接检验脊髓 CaMKIIa 是促进和维持 SCD 慢性疼痛表现的分子机制这一假设，我们将使用化学、小干扰 RNA (siRNA) 和基因敲除方法进行抑制 CaMKIIa 的药理学研究。我们建议在这些药理学研究中测试一种临床使用的口服药物，我们发现该药物是一种 CaMKII 抑制剂，可以减少炎症和神经性疼痛模型中的疼痛行为。我们的第二个策略是进行一项试点转化研究，通过定量感觉测试和计算机化自我报告工具来表征人类的感觉疼痛，以确定这种 CaMKII 抑制剂的安全问题和临床潜力。该提案的意义在于，它可能最终导致针对 CaMKII 途径的药物干预。 （摘要完）