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: 9280774
• 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/9280774
• 关键词: Affect; Afghanistan; Alternative Splicing; Animal Experiments; Anti-Inflammatory Agents; Anti-inflammatory; Apoptosis; Apoptotic; Astrocytes; Astrocytosis; Atrial Natriuretic Factor; Attenuated; Behavior; Behavior assessment; Behavioral; Biochemical Process; Blood; Blood Cells; Bone Marrow; Bone Marrow Transplantation; Brain; Brain Injuries; Brain Stem; Brain natriuretic peptide; Brain region; Brain-Derived Neurotrophic Factor; CSF3 gene; Cell Culture Techniques; Cell Death; Cells; Cerebrovascular system; Chronic; Clinical Research; Complex; Contralateral; Devices; Dose; Endothelial Cells; Ensure; GDNF gene; Generations; Goals; Granulocyte Colony-Stimulating Factor; Green Fluorescent Proteins; Growth Factor; Hippocampus (Brain); Hour; Immunofluorescence Immunologic; Infiltration; Injury; Investigation; Ipsilateral; Iraq; Knock-out; Label; Lesion; Leukocytes; Light; Measures; Mediating; Mediation; Methods; Microglia; Military Personnel; Molecular; Molecular Analysis; Motor; Mus; Neuraxis; Neuroimmune system; Neurologic; Neurologic Deficit; Neurons; Operative Surgical Procedures; Penetration; Pharmaceutical Preparations; Phase; Phenotype; Physiological; Play; Prevention; Procedures; Process; Receptor Cell; Recording of previous events; Recovery; Regenerative response; Reporting; Research; Rodent Model; Role; Rotarod Performance Test; Signal Pathway; Signal Transduction; Stem cells; Stroke; Subarachnoid Hemorrhage; Survivors; Terrorism; Testing; Time; Tissue Sample; Translations; Traumatic Brain Injury; Traumatic Brain Injury recovery; Up-Regulation; Vascular Endothelial Cell; Water; Whole-Body Irradiation; angiogenesis; brain cell; brain repair; brain tissue; chemokine; chemokine receptor; controlled cortical impact; cytokine; design; disability; high risk; injured; intravenous administration; monocyte; monocyte chemoattractant protein 1 receptor; motor deficit; mouse model; nerve stem cell; neurogenesis; neuroinflammation; neurological recovery; neuron apoptosis; neuron loss; neuronal survival; neurotransmission; neurotrophic factor; novel strategies; programs; public health relevance; receptor; regenerative; repaired; response; standard care; vasogenic edema

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）是一个复杂的过程，包括三个重叠阶段：a）初级损伤，b）继发性损伤和c）再生反应。全身给药粒细胞 - 固体刺激因子（G-CSF）代表了一种增强大脑自我修复的新方法，尤其是在次生和再生阶段。该研究计划的具体目的旨在检验A）G-CSF间接增强脑修复的假设，从而通过促进骨髓来源的细胞（BMDC）来增强脑修复，从而调节神经炎症过程并分泌营养因素； b）G-CSF直接与其神经元细胞受体相互作用，以触发细胞内信号传导级联反应，从而减少细胞死亡并促进神经发生。 AIM 1将确定G-CSF的最佳剂量和小鼠TBI后的行为恢复时间道路。非辐照小鼠将用于确保G-CSF的影响不会被全身照射和骨髓移植（BMT）混淆。在基线，TBI后3、7和14天，将评估MOTOR功能（偏置挥杆活动； Rotarod）和行为（水迷宫）终点。次要终点将测量a）病变体积，b）小胶质细胞增多和星形胶质细胞增多症的程度以及c）细胞因子的大脑区域水平。目标2a。为了评估TBI触发并由G-CSF调节的BMDC动员的程度，将使用免疫荧光来确定绿色荧光蛋白（GFP+）BMDC的表型命运和分布BMDC的分布，以鉴定微胶质细胞和神经元型细胞co-Express GFP。 GFP+ BMDC的浸润时间将通过评估受伤后3、7和14天的TBI的总GFP+ Burnen同侧和对侧。目标2B。为了确定BMDC渗透到大脑中的程度增加了TBI恢复，将通过阻断单核细胞趋化因子信号传导或用趋化因子受体CCR2敲除小鼠的趋化因子信号传导或利用趋化因子信号传导或利用趋化因子信号的药物。增强的恢复目的地抑制BMDC动员将支持以下假设：G-CSF对神经细胞的直接作用起着主要作用。目的3。为了研究G-CSF作用对神经元细胞的直接影响，将在神经元细胞培养物中评估这些细胞因子对信号转导，凋亡和神经发生的分子影响。该分析的结果将与从用G-CSF或媒介物处理的TBI大脑中解剖的组织样品中信号转导和抗凋亡的分子分析进行比较。方法：将生成嵌合小鼠，该嵌合小鼠携带GFP BMDC允许跟踪BMDC的分布和表型命运，这些BMDC在TBI后浸润大脑。手术：TBI将通过 气动驱动的受控皮质冲击（CCI）设备对小鼠。行为评估：运动不对称（EBST），rotarod测试和水迷宫（MWM）的分析。终点：a）行为变化； b）病变体积的变化； c）通过双标记程序评估的大脑中GFP+ BMDC的范围，分布和表型命运； d）大脑区域中细胞因子谱的变化； e）信号转导（PKC-）的变化，BCl2。预期结果：G-CSF将调节BMDCS浸润并增强行为缺陷的恢复。神经系统缺陷的改善将表明与行为的组合有关，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）直接作用于其神经元细胞受体的G-CSF触发的抗凋亡信号传导的上调。

项目成果

Hippocampal neurogenesis and the brain repair response to brief stereotaxic insertion of a microneedle.

• DOI: 10.1155/2013/205878
• 发表时间: 2013
• 期刊: Stem cells international
• 影响因子: 4.3
• 作者: Song S;Song S;Cao C;Lin X;Li K;Sava V;Sanchez-Ramos J
• 通讯作者: Sanchez-Ramos J

Detrimental effects of physical inactivity on neurogenesis.

缺乏身体活动对神经发生的不利影响。

• DOI: 10.4103/2394-8108.186278
• 发表时间: 2016-04
• 期刊: Brain circulation
• 影响因子: 1.9
• 作者: Lippert T;Watson N;Ji X;Yasuhara T;Date I;Kaneko Y;Tajiri N;Borlongan CV
• 通讯作者: Borlongan CV

Transient Microneedle Insertion into Hippocampus Triggers Neurogenesis and Decreases Amyloid Burden in a Mouse Model of Alzheimer's Disease.

瞬时微针插入海马体可触发阿尔茨海默病小鼠模型中的神经发生并减少淀粉样蛋白负担。

• DOI: 10.3727/096368916x691114
• 发表时间: 2016
• 期刊: Cell transplantation
• 影响因子: 3.3
• 作者: Song,Shijie;Kong,Xiaoyung;Sava,Vasyl;Cao,Chuanhai;Acosta,Sandra;Borlongan,Cesar;Sanchez-Ramos,Juan
• 通讯作者: Sanchez-Ramos,Juan