Regulation and Role of Ceramidase in Inflammation

神经酰胺酶在炎症中的调节和作用

• 批准号: 8195563
• 负责人: Lina M OBEID
• 金额: --
• 依托单位: RALPH H JOHNSON VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-10-01 至 2013-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8195563
• 关键词: Adverse effects; Animal Model; Animals; Anti-Inflammatory Agents; Anti-inflammatory; Antibodies; Arthritis; Automobile Driving; Binding; Cardiovascular system; Cell Line; Cell Survival; Cell model; Cells; Ceramidase; Ceramides; Chronic; Colitis; Complementary DNA; Data; Dinoprostone; Enzymes; Epithelial Cells; Family; Family member; Fibroblasts; Flax; Funding; G-Protein-Coupled Receptors; Generations; Genes; Goals; Health; Homologous Gene; Human; Human Cloning; Immunohistochemistry; Incidence; Inflammation; Inflammatory; Inflammatory Response; Knock-out; Laboratories; Lead; MAPK8 gene; Malignant Neoplasms; Mediating; Metabolic; Metabolism; Modality; Modeling; Mus; Normal tissue morphology; Pathway interactions; Phosphorylation; Principal Investigator; Process; Production; Proteins; Regulation; Relative (related person); Research; Rheumatoid Arthritis; Role; Saccharomyces cerevisiae; Sampling; Scheme; Small Interfering RNA; Sphingolipids; Sphingosine; Sphingosine-1-Phosphate Receptor; Stimulation of Cell Proliferation; Stimulus; Testing; Tissues; Tumor Necrosis Factor-alpha; Ulcerative Colitis; Veterans; angiogenesis; cell type; cyclooxygenase 2; cytokine; galactosylgalactosylglucosylceramidase; human tissue; in vivo; in vivo Model; inhibitor/antagonist; insight; macrophage; member; mouse model; novel; novel therapeutic intervention; patient population; programs; prototype; public health relevance; response; sphingosine 1-phosphate; sphingosine kinase; tool

DESCRIPTION (provided by applicant): The long-term goal of this project is to define the role of the ceramide metabolizing enzyme family of ceramidases in inflammation and to target these enzymes for novel anti-inflammatory therapy. The PI's laboratory has an established track record of expertise in sphingolipid metabolism and function. Studies from the previous funding period have led us into a novel exciting direction on the role and regulation of ceramidases in and their metabolic products in inflammation. Ceramidases breakdown ceramide to generate sphingosine and sphingosine-1-phosphate (S1P) which in turn mediates several biologic activities, including inflammatory responses. In this competing renewal we have compelling new data, whereby we have implicated this pathway as a key regulator of cyclooxygenase-2 (COX-2) expression and prostaglandin E2 (PGE2) production. In addition we find that at least one ceramidase is involved in the TNF inflammatory response. This proposal will therefore, test the hypothesis that ceramidases regulate cytokine-mediated chronic inflammation, and that inhibiting this ceramidase activity inhibits inflammatory responses. To test this hypothesis we propose the following aims: 1) Determine which ceramidases are regulated in inflammation and determine the mechanisms of this regulation. This will be done by determining which of the ceramidases (acid, neutral and alkaline) is/are upregulated in inflammation and studying the mechanisms of this regulation in cell models of inflammation, and by determining the expression and cell-type distribution of ceramidases in inflammatory tissues from humans and from animal models of inflammation. 2) Establish the function of ceramidases in regulation of inflammation and determine the mechanisms of action of their products. This will be done by demonstrating that ceramidases and their metabolic products have a significant role in regulating inflammation by evaluating the effect of over expression of ceramidases in cells and in vivo models of inflammation, and determining the mechanisms by which ceramidases regulate inflammatory pathways (S1P, NF-KB, ERKs, COX-2). 3) Determine the relative contribution of each of the ceramidases in inflammation and dissect the mechanisms involved. This will be done by blocking ceramidase activity in cells and in vivo using small interfering RNA to the different ceramidases. In addition we will test different compounds that we synthesized for their ability to inhibit ceramidases in cells and in vivo models of inflammation. We will also test if ceramidase K/O mice are protected from inflammation in a model of IBD colitis and TNF-induced arthritis. These studies will enable us to gain important insight into the role of the ceramidase pathway in inflammatory responses and may also provide novel therapeutic approaches to inflammation. PUBLIC HEALTH RELEVANCE: NARRATIVE. Relevance to Veterans Health. The VA patient population has a high incidence of chronic inflammatory conditions, in particular, arthritis and colitis. These conditions are difficult to treat and generally are not sensitive to many available modalities of treatment. Moreover, chronic inflammatory conditions are increasingly thought to lead to cancer. Tumor necrosis factor (TNF) and COX-2 are implicated in most of these chronic inflammatory conditions and recent highly effective anti-inflammatory therapy is geared at blocking their action. However, these anti-inflammatory agents have proven to have cardiovascular and other side effects. Our preliminary studies demonstrate that sphingolipid molecules are intermediates in the action of TNF on COX-2. We will therefore utilize models of rheumatoid arthritis and inflammatory colitis to study the sphingolipid regulating enzymes ceramidases in these models of inflammation and target these enzymes for novel therapies in the treatment of chronic inflammation.

描述（由申请人提供）： 该项目的长期目标是定义神经酰胺代谢酶家族在炎症中的作用，并将这些酶靶向新的抗炎疗法。 PI的实验室在鞘脂代谢和功能方面具有既定的专业知识记录。从上一期资金期开始的研究使我们朝着神经酰胺酶中的作用和调节及其代谢产物在炎症中的作用和调节迈出了一个新的激动人心的方向。神经酰胺酶分解神经酰胺会产生鞘氨醇和鞘氨醇1-磷酸盐（S1P），进而介导了几种生物学活性，包括炎症反应。在这种竞争的更新中，我们有了引人注目的新数据，因此我们将这一途径牵涉到环氧合酶-2（COX-2）表达和前列腺素E2（PGE2）生产的关键调节剂。此外，我们发现至少有一个神经酶参与TNF炎症反应。因此，该建议将检验以下假设：神经酶调节细胞因子介导的慢性炎症，并抑制这种神经酰胺酶活性抑制炎症反应。 为了检验该假设，我们提出以下目的：1）确定哪些神经酶在炎症中受到调节，并确定该调节的机制。这将通过确定炎症中的哪种神经酰胺酶（酸，中性和碱）在炎症中上调，并研究这种调节的炎症细胞模型的机制，以及确定炎症性神经酰胺酶的表达和细胞类型来自人类的组织和炎症的动物模型。 2）确定神经酶在调节炎症中的功能，并确定其产物的作用机理。这将通过证明神经酶及其代谢产物在调节炎症中具有重要作用，通过评估神经酰胺酶在细胞和体内炎症模型中的过度表达的影响，并确定神经酰胺酶调节炎症途径的机制（S1P，S1P，S1P，， NF-KB，ERKS，COX-2）。 3）确定每个神经酶在炎症中的相对贡献，并剖析涉及的机制。这将通过使用小型干扰RNA到不同的神经酶的细胞和体内阻断神经酰胺酶活性来完成。此外，我们将测试我们合成的不同化合物，以抑制细胞和体内炎症模型中的神经酰胺酶的能力。我们还将在IBD结肠炎和TNF诱导的关节炎模型中测试神经酰胺酶K/O小鼠免受炎症的保护。 这些研究将使我们能够对神经酶途径在炎症反应中的作用进行重要了解，并可能提供新颖的炎症治疗方法。 公共卫生相关性： 叙述。与退伍军人健康有关。 VA患者人群的慢性炎症疾病的发生率很高，特别是关节炎和结肠炎。这些疾病很难治疗，通常对许多可用的治疗方式不敏感。此外，越来越多地认为慢性炎症状况会导致癌症。肿瘤坏死因子（TNF）和COX-2在大多数这些慢性炎症条件下都涉及，并且最近高效的抗炎疗法旨在阻止其作用。但是，这些抗炎药已被证明具有心血管和其他副作用。我们的初步研究表明，在TNF对COX-2的作用中，鞘脂分子是中间体。因此，我们将利用类风湿关节炎和炎症性结肠炎模型研究这些炎症模型中的鞘脂调节神经酶酶，并靶向这些酶用于治疗慢性炎症的新型疗法。