Heparin on Hypoxic Pulmonary Hypertension and Remodeling

肝素对缺氧性肺动脉高压和重塑的影响

• 批准号: 7897871
• 负责人: CHARLES A HALES
• 金额: $ 44.86万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-07-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7897871
• 关键词: Acetylation; Alkanesulfonates; Anatomy; Animals; Anticoagulants; Antihypertensive Agents; Biochemical; Carbohydrates; Cavia; Cell Proliferation; Chemical Structure; Cleaved cell; Clinical; Core Protein; Cyclins; Cytosol; Data; Development; Disaccharides; Distal; Divorce; Down-Regulation; Endothelin; Epoprostenol; Extracellular Matrix; Family suidae; Glucosamine; Glycosaminoglycans; Grant; Growth; Heparin; Hexuronic Acids; Human; Hyperplasia; Hypertension; Hypertrophy; Hypoxia; In Vitro; Investigation; Lead; Length; Low-Molecular-Weight Heparin; Lung; Mediating; Modeling; Morbidity - disease rate; Mus; Nuclear Envelope; Pathway interactions; Phosphorylation; Phosphotransferases; Pre-Clinical Model; Principal Investigator; Proliferating; Property; Pulmonary Hypertension; Pulmonary Vascular Resistance; Pulmonary artery structure; Rattus; Rho Factor; Role; Side; Signal Pathway; Signal Transduction; Smooth Muscle Myocytes; Stimulus; Structure; Therapy Clinical Trials; Time; Translations; Up-Regulation; Vascular remodeling; Work; antiproliferative agents; cell growth; effective therapy; in vivo; mortality; programs; response; rho; success; sugar

DESCRIPTION (provided by applicant): Vascular remodeling with hypertrophy, hyperplasia and distal extension of pulmonary artery smooth muscle cells (PASMC) is a major component of the pulmonary vascular resistance in pulmonary hypertension. Several agents have become available to treat remodeling but none have been very satisfactory. Heparin is a normal constituent of the lung and is antiproliferative for PASMC. This grant continues our studies to understand how the cluster of varying glycosaminoglycans known as heparin work as an antiproliferative agent and to develop them, or their derivatives, into effective therapies for pulmonary hypertension. Unfractionated heparin contains a central protein core with many carbohydrate side chains which vary in content from batch to batch. We have found some of these batches to be much more antiproliferative for PASMC than others and to be better at inhibiting the development of hypoxia induced hypertension and remodeling in mice, rats, guinea pigs and now pigs. Low molecular weight (LMW) heparins are the carbohydrate side chains of heparin cleaved off the protein core and are not very antiproliferative. We have created more potent antiproliferative LMW heparins by adding bulky hexanoylated (H) or butanoylated (B) groups. We have also shown that heparin inhibits PASM proliferation by stimulating the cyclin kinase p27. We have preliminary evidence that heparin may do this by suppressing ERK and Rho/ROCK activity. With this background we plan the following specific aims: 1. Determine the structures in heparin's carbohydrate side chains which mediate the antiproliferative action on PASMC by: a. identifying the importance of the 2-O sulfonate groups on the hexuronic acid residues as well as the proportions of basic sugar residues, i.e. N- acetylation and N-sulfonation of glucosamine in the repeating disaccharide units necessary to give maximum antiproliferative activity, b. altering the chemical structure of our H and B LMW heparins to match features identified as enhancing the antiproliferative strength in step 1a, to make these LMW heparin derivatives even more potent, and c. increasing the length of the H and B LMW heparins to octa and decanoylated chains to try to achieve even greater antiproliferative potency. 2. Determine in a preclinical model if potent antiproliferative heparins or heparin derivatives can inhibit or reverse established pulmonary hypertension in hypoxic pigs. 3. Determine the mechanism by which heparin enhances p27 activity (ERK, Rho/ROCK; etc), build on our pilot data that strongly antiproliferative heparins inhibit RhoA/ROCK while weakly antiproliferative heparins do not, and verify our findings in heparin treated hypoxic mice.

描述（由申请人提供）：用肥大，增生和肺动脉平滑肌细胞的远端延伸（PASMC）是肺血管耐药性的主要成分。几种代理已经可以治疗重塑，但没有一个非常令人满意。肝素是肺的正常成分，对PASMC是抗增殖的。这项赠款继续我们的研究，以了解被称为肝素的不同糖胺聚糖簇如何作为抗增殖剂，并将其开发为有效的肺动脉高压疗法。未分离的肝素包含一个中心蛋白质核，其中有许多碳水化合物的侧链，从批处理到批量各不相同。我们发现，其中一些批次比其他批次比其他批次要比其他批次更具抗增殖性，并且可以更好地抑制低氧诱导的高血压和重塑的发展，而小鼠，大鼠，豚鼠和现在的猪。低分子量（LMW）肝素是肝素的碳水化合物侧链，脱离了蛋白质核心，不是很抗增殖。我们通过添加笨重的六烷酰基（H）或丁酰化（B）组来创建更有效的抗增生LMW肝素。我们还表明，肝素通过刺激细胞周期蛋白激酶p27抑制了帕斯的增殖。我们有初步证据表明肝素可以通过抑制ERK和RHO/ROCK活动来做到这一点。在此背景下，我们计划以下特定目的：1。确定肝素碳水化合物侧链中介导抗增殖作用的PASMC的结构。确定2-O磺酸盐基在己酸酸残基上的重要性以及基本糖残基的比例，即在重复二糖单元中葡萄糖的N-乙酰化和N-磺化，以获得最大的抗增殖活性所需的重复二糖单位。改变了我们的H和B LMW肝素的化学结构，以匹配该特征，该特征在第1A步中增强了抗增生强度，使这些LMW肝素衍生物更加有效，并且C。将H和B LMW肝素的长度增加到八八颗，并脱酰粘链，以实现更大的抗增殖效力。 2。在临床前模型中确定有效的抗增生性肝素或肝素衍生物是否可以抑制或反向低氧猪中建立的肺动脉高压。 3。确定肝素增强p27活性（ERK，RHO/ROCK; etc）的机制，建立在我们的飞行员数据上，这些数据强烈抗增生肝素会抑制RhoA/Rock，而弱抗增殖肝素则没有，并验证了我们在肝素治疗的低氧小鼠中的发现。