Reinforcing the Repair Response to Traumatic Brain Injury

加强对创伤性脑损伤的修复反应

• 批准号: 8546514
• 负责人: Shijie Song
• 金额: --
• 依托单位: JAMES A. HALEY VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8546514
• 关键词: Affect; Afghanistan; Alternative Splicing; Animal Experiments; Anti-Inflammatory Agents; Anti-inflammatory; Apoptosis; Apoptotic; Arachnoid mater; Astrocytes; Astrocytosis; Atrial Natriuretic Factor; Attenuated; Behavior; Behavior assessment; Behavioral; Biochemical Process; Blood; Blood Cells; Bone Marrow; Bone Marrow Transplantation; Brain; Brain Stem; Brain natriuretic peptide; Brain region; Brain-Derived Neurotrophic Factor; CSF3 gene; Cell Culture Techniques; Cell Death; Cells; Cerebrovascular Circulation; Chronic; Clinical Research; Complex; Contralateral; Devices; Dose; Edema; Endothelial Cells; Ensure; Fibroblast Growth Factor; GDNF gene; Generations; Goals; Granulocyte Colony-Stimulating Factor; Green Fluorescent Proteins; Growth Factor; Hemorrhage; Hippocampus (Brain); Hour; Immune system; Immunofluorescence Immunologic; Infiltration; Inflammatory; Injury; Investigation; Ipsilateral; Iraq; Knock-out; Label; Lesion; Leukocytes; Light; Measures; Mediating; Mediation; Methods; Microglia; Military Personnel; Molecular; Molecular Analysis; Motor; Mus; Neuraxis; Neurologic; Neurologic Deficit; Neurons; Operative Surgical Procedures; Penetration; Pharmaceutical Preparations; Phase; Physiological; Play; Prevention; Procedures; Process; Recording of previous events; Recovery; Reporting; Research; Rodent Model; Role; Rotarod Performance Test; Signal Pathway; Signal Transduction; Stem cells; Stroke; Survivors; Terrorism; Testing; Time; Tissue Sample; Translations; Traumatic Brain Injury; Up-Regulation; Vascular Endothelial Cell; Water; Whole-Body Irradiation; angiogenesis; brain cell; brain repair; brain tissue; cell growth; chemokine; chemokine receptor; controlled cortical impact; cytokine; design; disability; high risk; injured; intravenous administration; monocyte; mouse model; nerve stem cell; neurogenesis; neuron apoptosis; neuron loss; neuronal survival; neurotrophic factor; novel strategies; programs; public health relevance; receptor; regenerative; repaired; response; standard care

DESCRIPTION (provided by applicant): Traumatic brain injury (TBI) is a complex process encompassing three overlapping phases: a) primary injury, b) secondary injury and c) regenerative responses. Systemic administration of granulocyte-colony stimulating factor (G-CSF) represents a novel approach for reinforcing the brain's self-repair, especially during the secondary and regenerative phases. The Specific Aims of this research program are designed to test the hypotheses that a) G-CSF indirectly enhances brain repair by promoting infiltration of bone marrow-derived cells (BMDC) to brain, modulate neuro-inflammatory processes and secrete trophic factors; b) G-CSF directly interacts with its neural cells receptors to trigger intra-cellular signaling cascades that decrease cell death and promote neurogenesis. Aim 1 will determine the optimal dose of G-CSF and the recovery time-course of behavioral deficits after TBI in mice. Non-irradiated mice will be used to ensure the effects of G-CSF are not confounded by whole body irradiation and bone marrow transplantation (BMT). Motoric function (biased swing activity; rotarod) and behavioral (water maze) end-points will be assessed in mice at baseline, 3, 7, and 14 days after TBI. Secondary endpoints will measure a) lesion volume, b) extent of microgliosis and astrocytosis and c) brain regional levels of cytokines. Aim 2a. To assess the extent of BMDC mobilization triggered by TBI and modulated by G-CSF, the phenotypic fate and distribution of green fluorescent protein (GFP+) BMDC in chimeric mice brains will be determined using immunofluorescence to identify microglia, astrocytes, and neuron-like cells that co-express GFP. The time-course of infiltration of GFP+ BMDCs will be determined by assessing total GFP+ burden ipsilateral and contralateral to the TBI at 3, 7 and 14 days after injury. Aim 2b. To determine the extent to which BMDC penetration into brain is responsible for enhanced TBI recovery, the infiltration of BMDC into the central nervous system (CNS) will be attenuated or blocked with agents that block chemokine signaling to monocytes or utilization of mice with a knockout of the chemokine receptor CCR2. Enhanced recovery despite inhibition of BMDC mobilization will support the hypothesis that direct actions of G-CSF on neural cells play a major role. Aim 3. To investigate the direct effects of G-CSF action on neural cells, the molecular impact of these cytokines on signal transduction, apoptosis and neurogenesis will be assessed in neural cell cultures. Results from this analysis will be compared to molecular analyses of signal transduction and anti- apoptosis in tissue samples dissected from TBI brains treated with G-CSF or vehicle. Methods: Chimeric mice will be generated that harbor GFP BMDCs to permit tracking the distribution and phenotypic fate of BMDCs that infiltrate the brain after TBI. Surgery: TBI will be delivered with a pneumatically driven controlled cortical impact (CCI) device to mice. Behavioral Assessments: Analyses of motor asymmetry (EBST), rotarod test and Water Maze (MWM). Endpoints: a) changes in behavior; b) changes in lesion volume; c) extent, distribution and phenotypic fate of GFP+ BMDC in brain assessed by double-labeling procedures; d) changes in cytokine profiles in brain regions; e) changes in signal transduction (PKC-¿), Bcl2. Expected Results: G-CSF will modulate BMDCs infiltration and enhance recovery of behavioral deficits. Improvement of neurologic deficits will shown to be related to a combination of actions including a) changes in brain infiltration of BMDC; b) secretion of cytokines that promote neurogenesis; c) up-regulation of anti-apoptotic signaling triggered by G-CSF acting directly on its receptor in neural cells.

描述（由申请人提供）： 创伤性脑损伤（TBI颗粒）是一个复杂的过程，包含三个重叠阶段：a）原发性损伤，b）继发性损伤和c）细胞集落刺激因子（G-CSF）的全身施用代表了一种增强脑损伤的新方法。大脑的自我修复，特别是在次级和再生阶段，该研究计划的具体目标旨在测试以下假设：a) G-CSF 通过促进骨浸润间接增强大脑修复。骨髓源性细胞 (BMDC) 进入大脑，调节神经炎症过程并分泌营养因子；b) G-CSF 直接与其神经细胞受体相互作用，触发细胞内信号级联反应，从而减少细胞死亡并促进神经发生。确定 G-CSF 的最佳剂量以及小鼠 TBI 后行为缺陷的恢复时间过程将用于确保 G-CSF 的效果不会因全身照射而混淆。骨髓移植 (BMT) 将在基线、TBI 后 3、7 和 14 天对小鼠进行运动功能（偏向摆动活动；旋转）和行为（水迷宫）终点评估。体积，b) 小胶质细胞增生和星形细胞增多的程度，以及 c) 细胞因子的脑区域水平 目标 2a 评估由 TBI 触发并受调节的 BMDC 动员程度。 G-CSF、绿色荧光蛋白（GFP+）BMDC在嵌合体小鼠大脑中的表型命运和分布将使用免疫荧光来确定共表达GFP的小胶质细胞、星形胶质细胞和神经元样细胞的浸润时间过程。将通过在损伤后 3、7 和 14 天评估 TBI 同侧和对侧的 GFP+ 总负荷来确定 GFP+ BMDC，以确定目标 2b。 BMDC 渗入大脑在多大程度上促进了 TBI 恢复，使用阻断单核细胞趋化因子信号传导的药物或利用敲除趋化因子受体的小鼠，可以减弱或阻断 BMDC 渗入中枢神经系统 (CNS) CCR2。尽管抑制 BMDC 动员，但恢复增强将支持 G-CSF 对神经细胞的直接作用发挥主要作用的假设。 G-CSF 对神经细胞的作用，这些细胞因子对信号转导、细胞凋亡和神经发生的分子影响将在神经细胞培养物中进行评估，该分析的结果将与解剖组织样本中信号转导和抗细胞凋亡的分子分析进行比较。方法：将产生含有 GFP BMDC 的嵌合小鼠，以跟踪 TBI 手术后浸润大脑的 BMDC 的分布和表型命运。交付时带有 气动控制皮质冲击 (CCI) 装置对小鼠进行行为评估：运动不对称性分析 (EBST)、旋转测试和水迷宫 (MWM) 终点：a) 行为变化；b) 病变体积变化；通过双标记程序评估 GFP+ BMDC 在大脑中的分布和表型命运；e) 信号转导的变化； (PKC-¿)，Bcl2。 预期结果：G-CSF 将调节 BMDC 浸润并促进行为缺陷的恢复，这与一系列作用有关，包括 a) BMDC 脑浸润的变化； b) 促进神经发生的细胞因子的分泌；c) G-CSF 直接作用于神经细胞中的受体而上调抗凋亡信号。