Redor Pathways Regulating Nemeprotein Maturation in Asthma

Redor 通路调节哮喘中的 Neme 蛋白成熟

• 批准号: 9232190
• 负责人: DENNIS J STUEHR
• 金额: $ 31.39万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-15 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9232190
• 关键词: Address; Adrenergic beta-Agonists; Agonist; Air; Apoproteins; Asthma; Biological Models; Bronchoconstriction; Bronchodilation; Bronchodilator Agents; Cell Culture Techniques; Cells; Characteristics; Coculture Techniques; Data; Defect; Drug usage; Environment; Enzymes; Epithelial; Epithelial Cells; FDA approved; Glyceraldehyde-3-Phosphate Dehydrogenases; Health; Heme; Hemeproteins; Host Defense; Human; Individual; Inflammation; Liquid substance; Lung; Measures; Mediating; Metalloproteins; Modeling; Mus; Muscle Cells; Muscle function; Muscle relaxants; NOS1 gene; NOS2A gene; NOS3 gene; Nitrates; Nitric Oxide; Nitric Oxide Synthase; Nitrites; Oxidation-Reduction; Oxidoreductase; Pathway interactions; Patient Care; Patients; Pharmaceutical Preparations; Pharmacology; Play; Production; Prosthesis; Pulmonary Hypertension; Recycling; Refractory; Research; Respiratory physiology; Rest; Role; SKIL gene; Slice; Smooth Muscle; Soluble Guanylate Cyclase; Sulfhydryl Compounds; System; TXN gene; Testing; Translating; Up-Regulation; Vascular Smooth Muscle; airway epithelium; airway hyperresponsiveness; airway inflammation; asthmatic; asthmatic airway; base; clinically relevant; cytokine; desensitization; experimental study; heme a; improved; in vivo; macrophage; member; mouse model; novel; novel therapeutic intervention; prevent; programs; receptor; respiratory smooth muscle; response; restoration

ABSTRACT - PROJECT 3 Metalloproteins that contain a heme prosthetic group (hemeproteins) are essential to lung functions, including airway host defense and inflammation [inducible nitric oxide (NO) synthase (iNOS)], and bronchomotor tone [soluble guanylate cyclase (sGC)]. We recently discovered that NO inhibits heme insertion into apo-proteins by the S-nitrosyation of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which functions in heme transport and insertion. Recycling GAPDH is dependent on cellular redox and S-denitrosylase enzymes, i.e. GSNO reductase (GSNOR) or thioredoxin 1 (Trx1). These exciting findings and longstanding interactions with Program members in the study of asthma over the years made it a natural step to investigate whether the high- levels of NO in the asthmatic airway blocks hemeprotein maturation. Our preliminary results show that asthmatic airway epithelium escapes NO-mediated inhibition of heme insertion by upregulation of denitrosylase pathways, enabling the expression of active iNOS, but that airway smooth muscle cells are overwhelmed by NO at the levels produced in the asthmatic airway, with consequent loss of heme-replete sGC. New preliminary data generated in murine models of asthma and in the human precision cut lung slice model suggest that heme-independent sGC-activators are strikingly effective bronchodilators. Thus, we will test the hypothesis that thiol denitrosylase systems enable continuous airway epithelial NO production by iNOS in asthma by restoration of hemeprotein maturation, but that the NO overwhelms smooth muscle denitrosylase systems, compromising maturation & function of the sGC in smooth muscle, which results in bronchoconstriction and airway hyper-reactivity. We plan concurrent mechanistic studies and studies of the bronchodilator response to novel pharmacologic sGC agonists in (i) primary airway smooth muscle cells from healthy and asthmatic airways, (ii) precision cut human lung slice model and (iii) mouse models of asthma using genetically modified mice (including NOS1,2,&3 -/-, GSNOR-/- and smooth muscle specific sGC-/-). Also, we will study hemeprotein maturation in airway epithelial cells freshly obtained from healthy individuals and asthmatics with low or high FENO to help verify clinical relevance and potentially endotype sGC-agonist responders. sGC agonists will be tested in order to translate the discoveries to patient care soon. This research will provide the first information on hemeprotein maturation in healthy and asthmatic airway, and test connections between dysregulation of hemeprotein maturation and the bronchoconstriction and inflammation characteristic of asthma.

摘要--项目3 含有血红素修复基团的金属蛋白(血红素蛋白)对肺功能是必不可少的，包括 呼吸道宿主防御和炎症[诱导型一氧化氮合酶(INOS)]和支气管运动张力 [可溶性鸟苷环化酶(SGC)]。我们最近发现，NO通过以下途径抑制血红素插入载脂蛋白 参与血红素转运的甘油醛-3-磷酸脱氢酶的S亚硝化 和插入物。GAPDH的循环依赖于细胞内的氧化还原和S脱氮酶，即GSNO 还原酶(GSNOR)或硫氧还蛋白1(Trx1)。这些令人兴奋的发现和长期的互动 多年来从事哮喘研究的项目成员自然而然地调查了高水平的 哮喘呼吸道中的NO水平阻碍了血红蛋白的成熟。我们的初步结果显示， 哮喘气道上皮通过上调脱氮酶逃脱NO介导的血红素插入抑制 途径，使活跃的iNOS表达，但气道平滑肌细胞被 NO在哮喘呼吸道中产生的水平，从而导致富含血红素的sGC的损失。新的 在小鼠哮喘模型和人类精密切片肺切片模型中产生的初步数据 表明不依赖于血红素的sGC激活剂是非常有效的支气管扩张剂。因此，我们将测试 硫醇脱氮酶系统可持续诱导大鼠气道上皮细胞产生NO的假说 哮喘通过恢复血红蛋白成熟，但NO压倒了平滑肌脱氮酶 系统，损害SGC在平滑肌中的成熟和功能，从而导致 支气管收缩和呼吸道高反应性。我们计划同时进行机械学研究和对 支气管扩张剂对新型药物sGC激动剂的作用(I)原代培养的气道平滑肌细胞 健康和哮喘的呼吸道，(Ii)精确切割的人肺切片模型和(Iii)哮喘的小鼠模型 使用转基因小鼠(包括NOS1、2和3-/-、GSNOR-/-和平滑肌特异性sGC-/-)。另外， 我们将研究新鲜从健康个体获得的呼吸道上皮细胞中的血红蛋白成熟情况。 低FeNO或高FeNO哮喘患者有助于验证临床相关性和潜在的内型sGC激动剂 响应者。SGC激动剂将进行测试，以便很快将发现转化为患者护理。这 研究将提供关于健康和哮喘呼吸道中的血红蛋白成熟的第一个信息，并进行测试 血红蛋白成熟失调与支气管收缩和炎症的关系 以哮喘为特征的。