Redor Pathways Regulating Nemeprotein Maturation in Asthma

Redor 通路调节哮喘中的 Neme 蛋白成熟

基本信息

批准号：
9015473
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
项目类别：
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/9015473
关键词：
Address Adrenergic beta-Agonists Agonist Air 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 Figs - dietary Glyceraldehyde-3-Phosphate Dehydrogenases Health Heme Hemeproteins Host Defense Human Individual Inflammation Liquid substance Lung Measures Mediating Metalloproteins Modeling Mus Muscle function Muscle relaxants NOS1 gene NOS2A gene NOS3 gene Nitrates Nitric Oxide Nitrites Oxidation-Reduction Oxidoreductase Pathway interactions Patient Care Patients Pharmaceutical Preparations Play Production Prosthesis Protein S Pulmonary Hypertension Recycling Research Respiratory physiology Rest Role Slice Smooth Muscle Smooth Muscle Myocytes Soluble Guanylate Cyclase Sulfhydryl Compounds System TXN gene Testing Translating Up-Regulation Vascular Smooth Muscle Work airway epithelium airway hyperresponsiveness asthmatic asthmatic airway base clinically relevant cytokine heme a improved in vivo macrophage member mouse model novel novel therapeutic intervention prevent programs receptor research study respiratory smooth muscle response restoration

项目摘要

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 含有血红素假体（囊蛋白）的金属蛋白对于肺功能至关重要，包括气道宿主防御和炎症[诱导一氧化氮（NO）合酶（iNOS）]和支气管运动 [可溶性鸟苷酸环化酶（SGC）]。最近，我们发现没有抑制血红素的插入 3-磷酸甘油醛-3-磷酸脱氢酶（GAPDH）的S-硝化作用，在血红素转运中起作用和插入。回收GAPDH取决于细胞氧化还原和S-葡萄糖基酶，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与之互动多年来，哮喘研究中的计划成员使研究高度是否是自然而然的一步哮喘气道中的NO水平阻断了囊蛋白的成熟。我们的初步结果表明哮喘气道上皮通过上调脱诺罗斯利酶来逃脱无介导的血红素插入的抑制作用途径，使活动iNOS的表达能够表达，但是气道平滑肌细胞被在哮喘气道中产生的水平没有，因此损失了血红素复发的SGC。新的在哮喘的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与血红素无关的SGC激活剂是非常有效的支气管扩张剂。因此，我们将测试假设硫醇根硝基酶系统可实现连续气道上皮无iNOS的产生通过恢复血红蛋白成熟的哮喘，但没有压倒平滑肌根硝基化酶系统，损害平滑肌中SGC的成熟和功能，这导致支气管收缩和气道高反应性。我们计划并发的机械研究和研究支气管扩张剂对（i）主要气道平滑肌细胞中新型药理学SGC激动剂的反应健康和哮喘的气道，（ii）精确切割人肺切片模型和（iii）哮喘的小鼠模型使用转基因的小鼠（包括NOS1,2，＆3 - / - ，GSNOR - / - 和平滑肌特异性SGC - / - ）。还，我们将研究来自健康个体的新鲜获得的气道上皮细胞中的囊蛋白成熟，具有低Feno或高FENO的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，以便将发现转化为患者护理。这研究将提供有关健康和哮喘气道中血蛋白成熟的第一个信息，并测试血管蛋白成熟和支气管收缩和炎症之间的连接哮喘的特征。