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 含有血红素辅基的金属蛋白（血红素蛋白）对肺功能至关重要，包括 气道宿主防御和炎症[诱导型一氧化氮（NO）合酶（iNOS）]和支气管张力 [可溶性鸟苷酸环化酶（sGC）]。我们最近发现NO通过抑制血红素插入脱辅基蛋白， 在血红素转运中起作用的甘油醛-3-磷酸脱氢酶（GAPDH）的S-亚硝基化 和插入。GAPDH的再循环依赖于细胞氧化还原和S-脱硝酶，即GSNO 还原酶（GSNOR）或硫氧还蛋白1（Trx 1）。这些令人兴奋的发现和长期的互动， 多年来研究哮喘的项目成员很自然地调查了高血压是否是一种疾病， 哮喘气道中的NO水平阻断血红素蛋白成熟。我们的初步结果显示， 哮喘气道上皮细胞通过上调脱硝酶逃避NO介导的血红素插入抑制 通路，使活性iNOS的表达，但气道平滑肌细胞被淹没 哮喘气道中产生的NO水平，随之而来的是充满血红素的sGC的损失。新 在哮喘小鼠模型和人类精确切割肺切片模型中产生的初步数据 表明血红素非依赖性sGC激活剂是显著有效的支气管扩张剂。我们将测试 假设硫醇脱硝酶系统能够通过诱导型一氧化氮合酶持续产生气道上皮NO， 哮喘通过血红素蛋白成熟的恢复，但NO抑制平滑肌脱硝酶 系统，损害sGC在平滑肌中的成熟和功能，这导致 支气管收缩和气道高反应性。我们计划同时进行机理研究和 （i）来自以下的原发性气道平滑肌细胞中对新型药理学sGC激动剂的支气管扩张剂反应： 健康和哮喘气道，（ii）精确切割人肺切片模型和（iii）哮喘小鼠模型 使用遗传修饰的小鼠（包括NOS 1，2，&3-/-、GSNOR-/-和平滑肌特异性sGC-/-）。还有， 我们将研究从健康个体新鲜获得的气道上皮细胞中血红素蛋白的成熟， 低或高FENO的哮喘患者，以帮助验证临床相关性和潜在的内源性sGC激动剂 响应者。将对sGC激动剂进行测试，以便将这些发现很快转化为患者护理。这 这项研究将提供健康和哮喘气道中血红素蛋白成熟的第一个信息， 血红素蛋白成熟失调与支气管收缩和炎症的关系 哮喘的特征。