AIRWAY MYOCYTE PROLIFERATION AND LYSOSOMAL HYDROLASE

气道肌细胞增殖和溶酶体水解酶

• 批准号: 2468206
• 负责人: DUKHEE Betty LEW
• 金额: $ 14.49万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-02-15 至 2002-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2468206
• 关键词: antisense nucleic acid; beta N acetylhexosaminidase; beta glucuronidase; bronchospasm; carbohydrate receptor; cell proliferation; enzyme activity; enzyme induction /repression; enzyme inhibitors; enzyme mechanism; focal adhesion kinase; immunocytochemistry; immunofluorescence technique; immunoprecipitation; inositol phosphates; integrins; lysosomes; mannose; mitogen activated protein kinase; prostaglandin E; protein kinase C; respiratory muscles; smooth muscle; tissue /cell culture

DESCRIPTION (Adapted from the applicant's abstract): The long term objectives of this proposal are to investigate the mechanism(s) involved in airway smooth muscle cell proliferation induced by lysosomal hydrolases and to determine the contribution of lysosomal hydrolases in the pathogenesis of asthma. The overall hypothesis underlying this proposal is that lysosomal hydrolases promote hyperplasia/hypertrophy and subsequent airway hyperreactivity. The proposal is based on recent findings of a potent mitogenic action of mannosyl-rich glycoproteins including lysosomal hydrolases (beta-hexosaminidase A & B and beta-glucuronidase) on bovine airway smooth muscle cells. This mitogenic action is mediated by 175 kD mannose recognizing receptors (ASM-MR). Specific aims are: 1) To determine the contribution of p42 and p44 mitogen activated protein kinases (p42/44MAPK) and 70-kD ribosomal protein S6 kinase (p70S6k) in mannosyl-rich glycoprotein-induced mitogenesis in airway smooth muscle cells (ASMC). 2) To determine the contribution of p72Syk, and p125 focal adhesion kinase (p125FAK) in mannosyl-rich glycoprotein-induced mitogenesis in ASMC. 3) To investigate if airway smooth muscle mannose receptor (ASM-MR) and integrin(s) cooperate in transducing signal(s) in mannosyl-rich glycoprotein-induced mitogenesis. To accomplish specific aims 1 and 2, the activities of above kinases and cell proliferation in response to Hex and a mannosyl-rich neoglycoprotein in the presence and the absence of specific inhibitors, antisense-, sense-, and scrambled-oligonucleotides will be correlated. To accomplish specific aim 3, co-immunoprecipitation, cross linking analysis, immunocytochemistry, 35S-methionine metabolic labeling/immunoprecipitation analysis will be performed using specific antibodies to integrins that use the RGD recognition sequence. Integrin expression will be inhibited by multiple pulses of antisense-oligonucleotides and its effect will be examined in mannosyl-rich glycoprotein-induced mitogenesis. The proposed studies are intended to advance the knowledge of the mechanism(s) underlying airway smooth muscle proliferation.

描述(改编自申请人的摘要)：长期 这项建议的目的是调查涉及的机制(S) 溶酶体水解酶和血管紧张素转换酶诱导的气道平滑肌细胞增殖 目的：探讨溶酶体水解酶在糖尿病发病中的作用。 哮喘。支持这一建议的总体假设是溶酶体 水解酶促进增生/肥大和随后的呼吸道 高反应性。这项建议是基于最近一项强有力的 包括溶酶体在内的富含甘露糖糖蛋白的促有丝分裂作用 牛的水解酶(β-氨基己糖苷酶A和B和β-葡萄糖醛酸酶) 呼吸道平滑肌细胞。这种促有丝分裂作用是由175kD介导的 甘露糖识别受体(ASM-MR)。具体目标是：1)确定 P42和p44丝裂原活化蛋白激酶的作用 (p42/44MAPK)和70kD核糖体蛋白S6K(P70S6K)在富含甘露醇的组织中的表达 糖蛋白诱导气道平滑肌细胞(ASMC)有丝分裂。2) 确定p72Syk和p125粘着斑激酶的作用 (P125FAK)在富含甘露糖醇的糖蛋白诱导的ASMC有丝分裂中。3)至 探讨气道平滑肌甘露糖受体(ASM-MR)和 富含甘露糖醇的整合素(S)配合传导信号(S) 糖蛋白诱导的有丝分裂。为了实现具体目标1和2， Hex和a对上述酶活性和细胞增殖的影响 富含甘露糖醇的新糖蛋白的存在和缺失 抑制剂、反义、正义和杂乱寡核苷酸将是 相互关联。为实现特定目标3，免疫共沉淀，交叉 连接分析、免疫细胞化学、35S-蛋氨酸代谢 标记/免疫沉淀分析将使用特定的 使用RGD识别序列的整合素抗体。整合素 表达将被多个脉冲抑制 反义寡核苷酸及其在富含甘露糖醇中的作用将被检测 糖蛋白诱导的有丝分裂。拟议的研究旨在 提高对呼吸道平滑肌作用机制的认识(S) 扩散。