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: 8368962
• 负责人: Charles Lee Howe
• 金额: $ 7.95万
• 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/8368962
• 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; 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. PUBLIC HEALTH RELEVANCE: The proposed research is relevant to public health because it will provide new models for the analysis of leukocyte trafficking into target tissues such as the central nervous system. The ability to better characterize neutrophil and monocyte entry into the infected brain, for example, may lead to the discovery of novel therapeutic strategies that will impact a wide array of human diseases. Thus, the proposed research is relevant to the mission of the NIH because it will provide tools for the extension of fundamental knowledge that will alleviate or reduce the burden of human disease.

描述（由申请人提供）：脑和脊髓浸润炎性单核细胞和中性粒细胞参与多种神经系统疾病的发病、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，可以提供不同骨髓单核细胞群体的可变和重叠分辨率，但这些工具对于识别、跟踪和定量靶组织（如大脑）中的单核细胞和中性粒细胞并不理想。Graf开发的LysM-eGFP小鼠，其中只有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复杂的工具。与此同时，新兴但相互矛盾的文献表明，中性粒细胞和单核细胞募集到中枢神经系统是相当复杂的，依赖于各种趋化因子和趋化因子受体的相互作用。例如，一般来说，中性粒细胞运输可能依赖于通过CXCR2和CXCR3轴的信号传导，而单核细胞运输可能依赖于CCR2和CCR5受体。基于这一概念，并努力分析特定趋化因子受体在骨髓单核细胞运输到中枢神经系统中的作用，我们的第一个目标是