Targeting the BBB to treat CNS inflammation

靶向 BBB 治疗中枢神经系统炎症

基本信息

批准号：
8826187
负责人：
Robyn S Klein
金额：
$ 28.31万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-09-25 至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8826187
关键词：
Address Affect Animal Model Animals Autoimmune Diseases Autopsy Biological Markers Biological Preservation Biopsy Specimen Blood Blood - brain barrier anatomy Blood Vessels Brain CNS autoimmunity CXCL11 gene CXCL12 gene CXCR4 Receptors Cell Polarity Cells Cellularity Clinical Cues Deposition Diffusion Diffusion Magnetic Resonance Imaging Disease Drug Targeting Edema Encephalitis Endothelial Cells Endothelium Enhancing Lesion Exhibits Experimental Autoimmune Encephalomyelitis Experimental Designs Gadolinium Genetic Human Image Immune In Situ In Vitro Infiltration Inflammation Inflammatory Investigation Lead Lesion Leukocyte Trafficking Leukocytes Life Location Lymphocyte Magnetic Resonance Imaging Mediating Modality Modeling Molecular Movement Multiple Sclerosis Multiple Sclerosis Lesions Mus Myelin Neuraxis Outcome Pathogenesis Patients Permeability Process Publishing Receptor Signaling Relapse Role Signal Transduction Site Surface T-Lymphocyte Testing Th1 Cells Tissues Translations Transport Vesicles Vascular Permeabilities axon injury base cell motility chemokine cytokine gadolinium oxide human tissue imaging modality in vivo intravital imaging intravital microscopy membrane polarity migration monocyte neuroimaging neuroinflammation novel receptor research study spectrograph sphingosine 1-phosphate trafficking transcytosis two-photon

Address Affect Animal Model Animals Autoimmune Diseases Autopsy Biological Markers Biological Preservation Biopsy Specimen Blood Blood - brain barrier anatomy Blood Vessels Brain CNS autoimmunity CXCL11 gene CXCL12 gene CXCR4 Receptors Cell Polarity Cells Cellularity Clinical Cues Deposition Diffusion Diffusion Magnetic Resonance Imaging Disease Drug Targeting Edema Encephalitis Endothelial Cells Endothelium Enhancing Lesion Exhibits Experimental Autoimmune Encephalomyelitis Experimental Designs Gadolinium Genetic Human Image Immune In Situ In Vitro Infiltration Inflammation Inflammatory Investigation Lead Lesion Leukocyte Trafficking Leukocytes Life Location Lymphocyte Magnetic Resonance Imaging Mediating Modality Modeling Molecular Movement Multiple Sclerosis Multiple Sclerosis Lesions Mus Myelin Neuraxis Outcome Pathogenesis Patients Permeability Process Publishing Receptor Signaling Relapse Role Signal Transduction Site Surface T-Lymphocyte Testing Th1 Cells Tissues Translations Transport Vesicles Vascular Permeabilities axon injury base cell motility chemokine cytokine gadolinium oxide human tissue imaging modality in vivo intravital imaging intravital microscopy membrane polarity migration monocyte neuroimaging neuroinflammation novel receptor research study spectrograph sphingosine 1-phosphate trafficking transcytosis two-photon

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项目摘要

Project 2 will determine how targeting molecules that regulate permeability and polarity at central nervous system (CNS) endothelial barriers impacts neuroinflammation, as assessed via novel neuroimaging modalities with murine models of multiple sclerosis (MS). We have shown that endothelium in MS lesions exhibits altered polarity, displaying the abluminal chemokine CXCL12 aberrantly along luminal surfaces. Abluminal CXCL12 normally serves to limit leukocyte entry whereas luminal expression of CXCL12 is associated with increased activation of its signaling receptor CXCR4 on infiltrating leukocytes. We showed that the scavenging CXCL12 receptor, CXCR7 (also expressed by CNS endothelium), is a critical regulator of leukocyte entry via internalization of CXCL12 from abluminal surfaces. In preliminary studies, we have also found that sphingosine 1-phosphate (S1P) signaling via S1P2 disturbs abluminal expression of CXCL12, which then disrupts immune privilege. Thus, mice with targeted deletion of S1P2 or administered a specific antagonist, JTE-013, show reduced migration of leukocytes into the CNS parenchyma during EAE. We hypothesize that alterations in membrane polarity of CXCL12 at CNS endothelial barriers promote leukocyte capture and migration into the CNS, contributing to the establishment of disease cycles in relapsing-remitting forms of CNS autoimmunity. We have identified several molecules that regulate this process, leading to reversal of apicobasal expression of the localizing cue, CXCL12. In this proposal, we will examine the roles of CXCR7 and S1P2 in lymphocyte trafficking and inflammation in CNS autoimmune disease using novel imaging modalities including two-photon intravital microscopy and longitudinally using Diffusion Basis Spectrum Imaging (DBSI), which distinguishes and quantitates cellularity and edema in live mice (see Project 1) and examination of autopsied human MS patient and control CNS tissues. We expect DBSI to be more sensitive than Gd enhancement to inflammation, including in the setting of less profound blood-brain barrier alterations. To determine the relationships between endothelial and immune cell (Th1 versus Th17) interactions and loss of BBB integrity, we will directly compare results utilizing intravital imaging to DBSI and Gd enhancement in the same animals. To address mechanisms by which endothelial cell polarity and localizing cues are induced by interactions with inflammatory cells, we will utilize in vitro and in situ approaches with human tissues. Thus, Aim 1 will determine whether CXCR7-mediated internalization of CXCL12 requires infiltration with Th1 versus Th17 cells during EAE. Aim 2 will determine whether S1P2- mediated reversal of endothelial cell polarity leads to increased T cell capture and entry during EAE, and Aim 3 will examine the relationship between CXCR7 and S1PR2 activation and T cell migration at the BBB in MS. The experimental design of Project 2 will define how DBSI determined cellularity and edema changes responding to various degrees of BBB abnormality and cell infiltration in EAE mice.

项目2将确定如何靶向调节渗透率和极性的分子中枢神经系统（CNS）内皮屏障会影响神经炎症，如通过具有多发性硬化症（MS）的鼠模型（MS）的新型神经影像模式。我们已经显示了 MS病变中的内皮表现出改变的极性，显示了空白趋化因子 CXCL12沿腔表面异常。 Abluminal CXCL12通常用于限制白细胞进入，而CXCL12的腔表达与激活增加有关其信号传导受体CXCR4在浸润的白细胞上。我们证明了清除 CXCL12受体，CXCR7（也由CNS内皮表达），是一个关键调节剂白细胞通过从空白表面对CXCL12的内在化进入。在初步研究中，我们还发现，通过S1P2的1-磷酸盐（S1P）信号传导干扰 CXCL12的表达，然后破坏免疫特权。因此，具有目标缺失的小鼠 S1P2或管理特定的拮抗剂JTE-013显示白细胞迁移降低 EAE期间进入CNS实质。我们假设是膜极性的改变中枢神经系统内皮屏障的CXCL12促进白细胞捕获并迁移到中枢神经系统中，在CNS的复发形式中建立疾病周期自身免疫性。我们已经确定了几个调节此过程的分子，导致本地化提示的丙基巴斯表达的反转，CXCL12。在此提案中，我们将检查CXCR7和S1P2在CNS中淋巴细胞运输和炎症中的作用使用新型成像方式（包括两光插入）的自身免疫性疾病使用扩散基频谱成像（DBSI）进行显微镜和纵向纵向区分和量化活小鼠的细胞和水肿（请参阅项目1）和检查尸体尸体MS患者和对照中枢神经系统组织的检查。我们希望DBSI对炎症的增强比GD增强更敏感，包括设置不太深刻的血脑屏障改变。确定内皮和免疫细胞（TH1与TH17）相互作用以及BBB完整性的丧失，我们将直接比较利用插入成像与DBSI和GD增强的结果动物。解决内皮细胞极性和定位提示的机制通过与炎症细胞相互作用引起的，我们将在体外和原位接近人体组织。因此，AIM 1将确定CXCR7介导的CXCL12的内在化是否在EAE期间，需要用TH1与Th17细胞浸润。 AIM 2将确定S1P2-是否是否内皮细胞极性的介导的逆转导致T细胞捕获和进入期间增加 EAE和AIM 3将检查CXCR7和S1PR2激活与T细胞之间的关系 MS的BBB迁移。项目2的实验设计将定义DBSI 确定的细胞性和水肿变化响应各种程度的BBB异常 EAE小鼠中的细胞浸润。