Transport of TNFalpha across the BBB

TNFα 跨 BBB 的转运

• 批准号: 7342451
• 负责人: Weihong Pan
• 金额: $ 22.56万
• 依托单位: LSU PENNINGTON BIOMEDICAL RESEARCH CTR
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7342451
• 关键词: Albumins; Antibodies; Autoradiography; Behavior; Biological Assay; Blood; Blood Vessels; Brain; Chest; Clinical; Co-Immunoprecipitations; Cytoskeletal Proteins; Data; Detection; Disruption; Dose; Drug Delivery Systems; Endocytosis; Endothelial Cells; Evoked Potentials; Extracellular Domain; Fc Receptor; Goals; Growth Associated Protein 43; Immunoblotting; Immunoprecipitation; Knockout Mice; Knowledge; Label; Lead; Left; Literature; Measures; Mediating; Molecular Chaperones; Mus; Mutation; Natural regeneration; Nervous System Physiology; Pan Genus; Patients; Peripheral; Permeability; Pharmacological Treatment; Phosphoproteins; Phosphorylation; Play; Polymerase Chain Reaction; Process; Proteomics; Recovery; Recovery of Function; Research; Research Project Grants; Rest; Role; Side; Signal Transduction; Sorting - Cell Movement; Spinal Cord; Spinal cord injury; System; Testing; Therapeutic; Time; Tissues; Transfection; Transport Process; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Up-Regulation; Western Blotting; axon regeneration; base; behavior test; capillary; comparative; cytokine; cytotoxic; functional outcomes; genetic regulatory protein; human MPP1 protein; human TNF protein; immunoreactivity; improved; lymphotoxin beta; mouse model; neurofilament; novel; permissiveness; programs; receptor; receptor upregulation; response; spinal cord regeneration; trafficking; transcriptional coactivator p75; transcytosis; uptake

DESCRIPTION (provided by applicant): The blood-brain and blood-spinal cord barriers (BBB/BSCB), lying between the CNS (brain and spinal cord) and its supplying capillary blood vessels, provide an immense interface for interaction and exchange of information between the CNS and the rest of the body. We have shown that the BSCB is not static or physically passive, but undergoes dynamic changes, to regulate the availability of the cytokine tumor necrosis factor a (TNFa) from blood to the CNS. In the proposed studies, we will test the hypothesis that TNFa, after being transported across the BBB/BSCB, can facilitate functional recovery after spinal cord injury (SCI). We will also characterize the mechanisms by which this specific transport can be regulated. To determine the functional implications of TNFe transport, we will first show that spinal cord uptake of radioactively labeled TNFe will increase specifically in a modified mouse model of thoracic hemisection, and that this increase will not coincide with passive disruption of the barrier (measured by increased paracellular permeability of albumin). We will then determine the effects of TNFa treatment by evaluating locomotor behavior, intraspinal conduction of evoked potentials, and histological evidence of long tract regeneration. We expect that small doses of TNFa will facilitate recovery whereas large doses will worsen the deficits resulting from SCI. To identify the roles of p55- and p75-receptors, we will study TNFa transport in the presence of receptor antibodies and in the receptor knockout mice. We predict that increased spinal cord uptake of TNFe after SCI will be dependent upon upregulation of the receptors, as determined by Western blot and quantitative PCR analysis. Since transcytosis of a cytokine across the BBB/BSCB is a complicated process involving not only the receptors but also other regulatory proteins, we will identify novel transport regulatory proteins by co-immunoprecipitation, comparative proteomics, transfection and transport assays. We predict that the transport process will be regulated by the receptors, vesicular and cytoskeletal proteins, and chaperons in different phosphorylation states. By completing the proposed study, we will better understand the dual roles of TNFa in SCI and in its transport process involving the regulatory proteins. These transporters could be novel drug targets and therefore provide promising therapeutic potential.

描述(申请人提供)：血-脑和血-脊髓屏障(BBB/BSCB)位于中枢神经系统(脑和脊髓)及其供应的毛细血管之间，为中枢神经系统和身体其他部分之间的互动和信息交换提供了一个巨大的接口。我们已经证明，BSCB不是静态的或身体上的被动的，而是经历动态的变化，以调节细胞因子肿瘤坏死因子a(TNFa)从血液到中枢神经系统的可用性。在拟议的研究中，我们将检验这样的假设，即TNFa通过BBB/BSCB转运后，可以促进脊髓损伤(SCI)后的功能恢复。我们还将描述这一特定运输受到管制的机制。为了确定TNFe转运的功能意义，我们将首先证明，在改良的小鼠胸部半横断模型中，脊髓对放射性标记的TNFe的摄取将特异性地增加，并且这种增加不会与被动破坏屏障(通过增加白蛋白的细胞旁通透性来衡量)相一致。然后，我们将通过评估运动行为、诱发电位的椎管内传导和长管再生的组织学证据来确定TNFa治疗的效果。我们预计，小剂量的TNFa将促进康复，而大剂量的TNFa将加剧脊髓损伤造成的赤字。为了确定p55和p75受体的作用，我们将在受体抗体存在的情况下和受体敲除小鼠中研究TNFa的转运。我们预测脊髓损伤后脊髓对TNFe摄取的增加将依赖于受体的上调，正如Western印迹和定量PCR分析所确定的那样。由于细胞因子跨过BBB/BSCB是一个复杂的过程，不仅涉及受体，还涉及其他调节蛋白，因此我们将通过免疫共沉淀、比较蛋白质组学、转染和运输试验来寻找新的运输调节蛋白。我们预测，转运过程将受到受体、囊泡和细胞骨架蛋白以及处于不同磷酸化状态的伴侣的调节。通过完成拟议的研究，我们将更好地了解TNFa在脊髓损伤中的双重作用及其在涉及调节蛋白的运输过程中的双重作用。这些转运蛋白可能是新的药物靶点，因此提供了很有前途的治疗潜力。