Imaging NFkB Activity in Relation to Animal Behavior in Peripheral Neuropathy

描述（由申请方提供）：椎间盘突出（IVD）可能与神经根压迫和生化刺激相关，导致炎性细胞浸润和相关疼痛或神经根病的产生。肿瘤坏死因子-α（TNF α）是一种促炎细胞因子，被认为是椎间盘突出中神经根病和神经性疼痛的早期介质。TNF α诱导的炎症受转录因子NF-kB调节，数据显示NF-kB拮抗作用可减少外周神经损伤后的伤害性疼痛。这项NRSA奖学金提出的主要假设是，通过非侵入性，纵向体内成像确定的区域NF-kB活性与周围神经损伤模型中啮齿动物的功能和疼痛相关行为有直接关系。第二个假设是在神经损伤区域局部抑制NF-kB可以改变疼痛相关行为。在目的1中，将在含有荧光素酶NF-κ B报告基因的转基因小鼠（NF-κ B-RE-Luc）中开发坐骨神经的慢性压迫性损伤（CCI）模型。神经系统中的局部NF-kB活性将通过体内发光成像，沿着时间匹配的功能测量（例如，步态）、疼痛相关行为（机械和热超敏反应、转轮活动）和炎性细胞浸润的表征。功能、疼痛和炎症细胞群的测量将与CCI和假手术对照中的局部NF-kB相关，以评估NF-kB对症状和功能障碍的贡献。在目标2中，姜黄素作为NF-κ B抑制剂的治疗效果将在CCI后通过非侵入性体内成像和功能和疼痛相关行为的标记物进行评估。在NFkB-RE-Luc转基因小鼠中CCI后，将姜黄素贮库神经周递送至坐骨神经，并且将通过体内发光成像纵向追踪小鼠的NF-kB活性。在CCI和神经周递送特异性NF-κ B抑制剂SC 514后，将类似地研究单独的一组动物。将在所有时间获得功能、疼痛行为和炎性细胞表征的测量，并检查两种模型中与NF-κ B活性的相关性，以评估局部姜黄素递送拮抗NF-κ B活性的能力，以及与CCI相关的症状和功能障碍。申请人将与申办者和指导团队合作，学习最先进的体内成像，步态分析和疼痛评估的新技术，这将促进申请人的职业生涯以及肌肉骨骼疾病研究的成像和功能生物标志物的翻译。申请人将获得更广泛的研究方法，动物模型，赠款和手稿准备和负责任的研究行为的培训，这将为他在生物工程的学术生涯做好准备。

DESCRIPTION (provided by applicant): Herniation of the intervertebral disc (IVD) may be associated with nerve root compression and biochemical irritation that contributes to inflammatory cell infiltration and associated generation of pain or radiculopathy. Tumor necrosis factor-a (TNFa) is a pro-inflammatory cytokine that is thought to be an early mediator of radiculopathy and neuropathic pain in herniated IVD. TNFa-induced inflammation is regulated by the transcription factor NF-kB, with data showing that NF-kB antagonism can reduce nociceptive pain following peripheral nerve injury. The main hypothesis of work proposed in this NRSA Fellowship is that regional NF-kB activity, determined via noninvasive, longitudinal in vivo imaging, has a direct relationship to rodent function and pain-related behaviors in a model of peripheral nerve injury. The secondary hypothesis is that localized inhibition of NF-kB in the region of nerve injury can modify pain-related behaviors. In Aim 1, a model of chronic constriction injury (CCI) of the sciatic nerve will be developed in transgenic mice (NFkB-RE-Luc) containing a luciferase NF-kB reporter. Regional NF-kB activity in the nervous system will be monitored longitudinally and non-invasively via in vivo luminescence imaging, along with time-matched measures of function (e.g., gait), pain-related behaviors (mechanical and thermal hypersensitivity, running wheel activity), and characterization of the inflammatory cell infiltrate. Measures of function, pain, and inflammatory cell populations will be correlated with regional NF-kB in the CCI and sham-operated controls in order to evaluate the contributions of NF-kB to symptoms and dysfunction. In Aim 2, the therapeutic efficacy of curcumin as an NF-kB inhibitor will be evaluated following CCI via noninvasive, in vivo imaging and markers of function and pain-related behaviors. Curcumin depots will be delivered perineural to the sciatic nerve following CCI in NFkB-RE-Luc transgenic mice, and mice will be tracked longitudinally for NF-kB activity via in vivo luminescence imaging. A separate set of animals will be similarly studied following CCI and perineural delivery of the specific NF-kB inhibitor, SC514. Measures of function, pain behaviors, and inflammatory cell characterizations will be obtained at all times and examined for correlations with NF-kB activity in both models, in order to assess an ability for local curcumin delivery to antagonize NF-kB activity, as well as symptoms and dysfunction associated with CCI. The applicant will work with the Sponsor and mentoring team to learn new techniques in state-of-the-art in vivo imaging, gait analysis, and pain assessments that will advance both the applicant's career and the translation of imaging and functional biomarkers for the study of musculoskeletal disease. The applicant will obtain broader training in research methodology, animal models, grant and manuscript preparation and responsible conduct in research that will prepare him for an academic career in bioengineering.