Preclinical Evaluation of Combination Therapy of Rolipram and Minocycline for Arresting Secondary Injury Cascade After Traumatic Brain Injury

咯利普兰和米诺环素联合治疗阻止创伤性脑损伤后继发性损伤级联的临床前评价

• 批准号: 10057824
• 负责人: Teresa Ann Murray
• 金额: $ 36.76万
• 依托单位: LOUISIANA TECH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10057824
• 关键词: Acute; Animals; Antibodies; Anxiety; Axon; Behavior; Behavioral; Biological Assay; Biological Markers; Biosensor; Brain; Cations; Cell Survival; Cells; Chemicals; Combined Modality Therapy; Cyclic AMP; Data; Dendrites; Development; Diffuse Brain Injury; Dose; Drug Combinations; Elements; Emotional; Exhibits; External Capsule; Functional disorder; Glass; Glutamates; Glycolic-Lactic Acid Polyester; Goals; Homeostasis; Hour; Human; Image; Immunofluorescence Immunologic; Impairment; Implant; Inflammation; Inflammatory; Injury; Laboratory mice; Lead; Life; Long-Term Effects; Louisiana; Measures; Memory; Minocycline; Modeling; Molecular; Monitor; Morphology; Motor; Mus; Nausea; Nerve Degeneration; Neurologic; Neurologic Deficit; Neurons; Optics; Periodicity; Pharmaceutical Preparations; Physiological; Polymers; Process; Recovery; Reproducibility; Resolution; Rolipram; Secondary to; Signal Transduction; Stains; Swelling; System; Testing; Therapeutic; Therapeutic Effect; Time; Traumatic Brain Injury; Varicosity; Work; amphiphilicity; awake; axon injury; behavior test; cell injury; cognitive function; cognitive recovery; controlled cortical impact; cytokine; delta opioid receptor; excitotoxicity; extracellular; fluid percussion injury; gamma-Aminobutyric Acid; glutamatergic signaling; high risk; imaging study; improved; in vivo; in vivo imaging; in vivo optical imaging; indexing; injured; insight; lens; long term memory; molecular marker; motor function recovery; mouse model; multimodality; multiphoton microscopy; nanocarrier; novel; novel drug combination; optical imaging; preclinical evaluation; response; restoration; serial imaging; tool; treatment duration; treatment effect

PROJECT SUMMARY For most traumatic brain injuries (TBI), the moment that an impact occurs, there is surprisingly little apparent damage. Yet, within minutes of impact, endogenous responses in the brain amplify and perpetuate the damage manifold. This cascade of responses is called secondary injury, and it is thought that this process can lead to long-term neurological problems. Our goal is to quickly mitigate some of the contributing elements of secondary injury to reduce long-term damage and neurological impairments. We will use a unique combination of drugs, including noninvasive delivery of one drug to the brain using a recently-developed co-polymer, poly (lactide-co-glycolide)-graft-polyethylenimine (PgP) as a nanocarrier. Morphological, physiological, chemical, and behavioral markers of secondary injury will be recorded at several time points up to one month after experimental TBI in mice to determine if the drug combination reduces secondary injury at an earlier time point than either drug alone, and if it maintains this reduction through one month after injury. We will use a unique multimodal system to monitor effects comprised of a newly developed microwire biosensor for glutamate (GLU) and γ-aminobutyric acid (GABA) with a modified micro-prism implant to correlate the dynamics of GLU and GABA signaling and ongoing secondary cellular damage by observing the same cells over time. Our highly sensitive and selective GLU and GABA microbiosensor will be used to evaluate the effect of treatment on reducing excitotoxicity by measuring extracellular concentrations and release dynamics in the cortex, in real time, at 5 time points after TBI versus preinjury baseline. At the same time points, high-resolution multiphoton microscopy through a permanently-implanted, modified prism will be used to facilitate a vertical view of cortical layers 3-6, to monitor development of varicosities (swellings) on dendrites and axons, axonal undulations and retraction bulbs, and the disappearance of axons. Furthermore, by viewing the same cells over time, we will compare the ability of the drugs to resolve dendritic and axonal varicosities on cells exhibiting damage in previous imaging sessions. Therapeutic effects will also be evaluated using standard behavioral tests for motor coordination, exploration, and memory, and by using established assays and antibody staining and cytokine assays for molecular biomarkers of secondary injury. By comparing the results from our novel combination of optical imaging and real-time glutamate and GABA signaling with the results from well-established behavioral and molecular biomarker tests at matched time points, we will demonstrate the utility of this longitudinal, optical- biosensor system to quantify secondary damage and its resolution. Notably, this longitudinal approach will require fewer animals because each animal serves as its own control, reducing variability, and each is used at multiple time points. In addition, optical-biosensor data from injured, vehicle-treated mice will provide new insights for better understanding the cascade of secondary injury and neural degeneration after TBI. Furthermore, this new optical-biosensor system will become a valuable tool for evaluating other drugs and for optimizing therapeutic windows.

项目摘要 对于大多数创伤性脑损伤（TBI），发生影响时，显而易见的很少 损坏。 这种反应的级联被称为次要伤害，这是 长期神经问题。我们的目标是快速减轻 减少长期损害和神经系统障碍的次要伤害。 药物，将药物无创递送到大脑风格开发的共聚物Poly （乳酸 - 糖丝酯） - 植物 - 聚甲基亚胺（PGP）作为纳米载体。 以及继发性伤害的行为标记将在几个月后的几个时间点记录 小鼠的实验性TBI确定药物组合是否在较早的时间点减少了继发性损伤 比单独使用任何一种药物，如果它保持了修订后，则在受伤后一个月的修订。 多模式系统监测由新开发的谷氨酸微无生物传感器（GLU）组成的效果 具有改良的微质植入物的γ-氨基丁酸（GABA）与GLU的动力学相关 随着时间的推移，GABA信号传导和持续的次级地窖损害 敏感和选择性的GLU和GABA微生物传感器将用于评估评估评估的评估 通过测量细胞外浓度和释放皮质中的动力学来降低兴奋性毒性，实际上 时间，在TBI与临时基线后的5个时间点。 通过永久植入，修改的棱镜的显微镜将促进皮质的垂直视图 3-6层，以监测树突和轴突上的静脉曲张（肿胀）的发展 缩回灯泡和轴突的诱发。 比较药物解决在细胞上造成损害的树突状和轴突静脉曲张的能力 预知成像课程也将评估运动的行为测试 通过既定测定法和抗体启动和细胞因子的协调，探索和记忆 通过我们的新型组合的结果，对次要损伤的分子生物标志物的测定 光学成像和实时谷氨酸和GABA信号传导，其结果良好 和分子生物标志物测试在匹配的时间点，我们将证明这种纵向，光学 - 生物传感器系统量化次要损害并尤其是这种纵向方法。 需要更少的动物，因为每只动物都是自身的控制，降低可变性，并且每个动物都在 此外，多个时间点。 洞察力，以更好地了解TBI后的继发性损伤和神经变性的级联。 此外，这种新的光学生物传感器系统将成为以太药物的宝贵工具和 优化治疗窗口。