CaMKII as a Mechanism & Intervention Target for Sickle Cell Pain

CaMKII 作为一种机制

基本信息

批准号：
8322679
负责人：
Zaijie Jim Wang
金额：
$ 42.26万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2016-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8322679
关键词：
Acute Adult Adverse effects African Age Animals Behavior Biological Markers Biometry Birth Chemicals Chemistry Chronic Clinical Data Development Disease Effectiveness Formalin Freund&apos s Adjuvant Genetic Goals Heating Human Immunoblotting Immunohistochemistry 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

项目摘要

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）：我们的长期目标是促进了解镰状细胞疾病（SCD）疼痛的神经机制的知识，并开发有效的药理治疗。 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产生并应用于研究SCD。然而，直到最近才发表疼痛发现，该发现表明使用辐射热尾润滑测试，NY1DD小鼠的疼痛敏感性提高。令人惊讶的是，这些小鼠的疼痛依赖于年龄，大约有6周的发作时间。不幸的是，研究人员没有报告其他疼痛测量结果，也没有在伯克利镰刀转基因小鼠中定义疼痛表型，这是一种更严重的SCD形式的模型。在我们自己的初步研究中，我们在一系列用于研究动物中其他疼痛类型的疼痛测试中检查了伯克利镰刀小鼠和窝窝非弹药对照。在这些初步研究中，我们发现触觉异常性痛和热痛觉过敏的存在，并且脊柱Camkii的表达和活性上调，类似于我们在其他炎症性和神经性疼痛的小鼠模型中观察到的类似。在后一种模型中，我们已经确定了 Camkiia是导致持续疼痛的关键组成部分。我们观察到Camkiia突变小鼠没有发展脊柱神经结扎诱导的疼痛行为。 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高原。这些疼痛测试中的一些用于我们正在进行的人类对SC疼痛的研究（QST）。有意地，这些相似性将使我们能够从SCD转基因小鼠模型中解释发现，并考虑到其他小鼠疼痛和人类SCD疼痛的发现。我们将使用实时PCR，免疫印迹，免疫史学化学和酶促动力学方法来系统地检查Camkiia和Camkii在镰刀和控制小鼠中的表达和活性，并将这种潜在的生物标志物的变化与疼痛发作相关联。为了直接检验脊柱CAMKIIA是一种分子机制的假设，可促进和维持SCD中慢性疼痛的表现，我们将使用化学，小型干扰RNA（siRNA）和基因敲除方法进行药理学研究以抑制CAMKIIA。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，我们发现它是CAMKII抑制剂，并减少炎症和神经性疼痛模型中的疼痛行为。我们的次要策略是进行一项试验转化研究，以通过定量的感觉测试和计算机化的自我报告工具来表征人类的感觉疼痛来确定该CAMKII抑制剂的安全问题和临床潜力。该提议的意义在于，它最终可能导致针对CAMKII-Pathway的药理干预措施。（抽象的结尾）