New tools to study leukocyte infiltration into the CNS

研究白细胞浸润中枢神经系统的新工具

• 批准号: 8497757
• 负责人: Charles Lee Howe
• 金额: $ 7.67万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8497757
• 关键词: Acute; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Animal Model; Animals; Applications Grants; Backcrossings; Bone Marrow; Brain; Brain Injuries; Breeding; CCL2 gene; CCL3 gene; CCL4 gene; CCL7 gene; CCR5 gene; CXCL1 gene; CXCL10 gene; CXCL2 gene; CXCL9 gene; CXCR3 gene; Cell Lineage; Cells; Chemokine (C-C Motif) Receptor 5; Communities; Complex; Conflict (Psychology); Consultations; Corpus Callosum; Coupled; Craniotomy; Demyelinating Diseases; Development; Disease; Endoscopes; Epilepsy; Fiber; Fiber Optics; Fluorescence; Goals; Guidelines; Hippocampal Formation; Hippocampus (Brain); IL8RB gene; Image; Immune system; Immunology; Infection; Infiltration; Inflammatory; Inflammatory Infiltrate; Kinetics; Knock-out; Knockout Mice; Knowledge; Lead; Leukocyte Trafficking; Leukocytes; Life; Literature; Malignant Neoplasms; Mediator of activation protein; Methods; Microglia; Microscope; Mission; Modeling; Mus; National Institute of Neurological Disorders and Stroke; Natural regeneration; Nerve Degeneration; Neuraxis; Neurologic; Neurosciences; Neutrophil Infiltration; Pain; Pathogenesis; Population; Public Health; Publishing; Recruitment Activity; Relative (related person); Reporter; Research; Research Personnel; Resolution; Resources; Rice; Role; Signal Transduction; Site; Spatial Distribution; Spinal Cord; Spinal cord injury; Stroke; Structure; Surface; TMEV; Time; Tissues; Traumatic Brain Injury; United States National Institutes of Health; Universities; Virus; Work; addiction; alveus; base; cancer pain; chemokine; chemokine receptor; fimbria; hippocampal fissure; human disease; indexing; injury and repair; innovation; interest; intravital imaging; macrophage; monocyte; monocyte chemoattractant protein 1 receptor; mouse model; nervous system disorder; neuroimmunology; neuroinflammation; neutrophil; novel therapeutics; therapeutic development; tool; trafficking; two-photon

DESCRIPTION (provided by applicant): Brain-and spinal cord-infiltrating inflammatory monocytes and neutrophils contribute to pathogenesis, injury, and repair/regeneration in a wide array of neurologic diseases, including stroke, epilepsy, demyelinating disease, Alzheimer disease, ALS, cancer, pain, TBI, spinal cord injury, and infection. While a number of surface markers exist that provide variable and overlapping resolution of different myelomonocytic populations, these tools are suboptimal for identifying, tracking, and quantitating monocytes and neutrophils in target tissues such as the brain. The development of the LysM-eGFP mouse by Graf, in which only cells of myelomonocytic lineage express GFP, has provided a far more sophisticated tool for following monocytes and neutrophils. At the same time, a burgeoning but conflicted literature indicates that neutrophil and monocyte recruitment to the CNS is quite complex and dependent upon a variety of chemokines and chemokine receptor interactions. For example, in general, neutrophil trafficking may depend upon signaling through the CXCR2 and CXCR3 axis, while monocyte trafficking may depend upon CCR2 and CCR5 receptors. Based on this concept, and in an effort to dissect the role of specific chemokine receptors in the trafficking of myelomonocytic cells to the CNS, our first objective in this small grant proposal is to cross LysM-eGFP mice with mice that are homozygously deficient in CCR2, CCR5, CXCR2, or CXCR3. Our second objective is to characterize the kinetics and spatial distribution of myelomonocytic cells infiltrating the brain in mice infected with the Theiler's murine encephalomyelitis virus. To accomplish this objective, we intend to use a fiber optic-based fluorescence endoscope to acquire deep tissue images of GFP-positive neutrophil and inflammatory monocyte trafficking in live animals and determine whether chemokine receptor deficiency alters the trafficking of the cells. Our long- term goals are to use these four lines to identify the factors responsible for leukocyte infiltration into the CNS and to assess the temporal inter-relation between myelomonocytic cells by thwarting infiltration of one population (for example neutrophils via CXCR2 deficiency) and quantifying the infiltration of other populations (for example monocytes). We intend to use these mouse models to determine basic aspects of leukocyte trafficking into the brain in our specific virus model and to make these lines available to other investigators studying stroke, TBI, spinal cord injury, etc. This project is innovative because it will generate new mouse models for more carefully studying neutrophil and inflammatory monocyte trafficking into the CNS and because it employs a fiber optic microscope to observe the trafficking of these cells within deep brain structures in living animals. Our proposed project is significant because it is expected to provide tools that will resolve a number of conflicting concepts regarding the mechanisms of leukocyte trafficking to the CNS. By extending our knowledge of neutrophil and monocyte trafficking mechanisms, these tools have the potential to greatly impact the development of therapeutic strategies for ameliorating human disease.

描述（由申请人提供）：脑和脊髓浸润性炎性单核细胞和中性粒细胞有助于多种神经系统疾病的发病机制、损伤和修复/再生，包括中风、癫痫、脱髓鞘疾病、阿尔茨海默病、ALS、癌症、疼痛、TBI、脊髓损伤和感染。虽然存在许多表面标记物，其提供不同骨髓单核细胞群体的可变和重叠分辨率，但这些工具对于鉴定、追踪和定量靶组织（如脑）中的单核细胞和嗜中性粒细胞而言是次优的。Graf开发的LysM-eGFP小鼠（其中仅骨髓单核细胞谱系的细胞表达GFP）为跟踪单核细胞和中性粒细胞提供了更为复杂的工具。与此同时，一个新兴的，但矛盾的文献表明，中性粒细胞和单核细胞的招聘到中枢神经系统是相当复杂的，并依赖于各种趋化因子和趋化因子受体的相互作用。例如，通常，嗜中性粒细胞运输可能依赖于通过CXCR 2和CXCR 3轴的信号传导，而单核细胞运输可能依赖于CCR 2和CCR 5受体。基于这一概念，并在努力剖析特定的趋化因子受体在骨髓单核细胞向中枢神经系统的运输中的作用，我们在这个小的赠款提案中的第一个目标是 将LysM-eGFP小鼠与CCR 2、CCR 5、CXCR 2或CXCR 3同源缺陷的小鼠杂交。我们的第二个目标是描述感染Theiler小鼠脑脊髓炎病毒的小鼠中骨髓单核细胞浸润脑的动力学和空间分布。为了实现这一目标，我们打算使用基于光纤的荧光内窥镜，以获取活动物体内GFP阳性中性粒细胞和炎性单核细胞贩运的深层组织图像，并确定趋化因子受体缺乏是否改变了细胞的贩运。我们的长期目标是利用这四条线， 确定导致白细胞浸润到CNS中的因素，并评估时间 通过阻碍一个群体的浸润（例如通过CXCR 2缺陷的嗜中性粒细胞）和定量其他群体（例如单核细胞）的浸润来确定骨髓单核细胞之间的相互关系。我们打算使用这些小鼠模型来确定在我们的特定病毒模型中白细胞运输到大脑中的基本方面，并使这些细胞系可用于研究中风、TBI、脊髓损伤、该项目是创新性的，因为它将产生新的小鼠模型，用于更仔细地研究中性粒细胞和炎性单核细胞向CNS的运输，并且因为它采用光纤显微镜观察这些细胞在活体动物大脑深层结构中的运输。我们提出的项目是重要的，因为它有望提供工具，将解决一些相互矛盾的概念，白细胞运输到中枢神经系统的机制。通过扩展我们对中性粒细胞和单核细胞运输机制的了解，这些工具有可能极大地影响改善人类疾病的治疗策略的发展。