AIRWAY MYOCYTE PROLIFERATION AND LYSOSOMAL HYDROLASE

气道肌细胞增殖和溶酶体水解酶

• 批准号: 6351498
• 负责人: DUKHEE Betty LEW
• 金额: $ 15.82万
• 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 2003-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6351498
• 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 该提案的目标是调查涉及的机制 溶酶体水解酶诱导气道平滑肌细胞增殖 确定溶酶体水解酶在发病机制中的贡献 哮喘。 该提议的总体假设是溶酶体 水解酶促进增生/肥大以及随后的气道 过度反应。 该提案基于一项有效的最新发现 富含甘露糖基的糖蛋白（包括溶酶体）的促有丝分裂作用 牛水解酶（β-己糖胺酶 A 和 B 以及 β-葡萄糖醛酸酶） 气道平滑肌细胞。 这种有丝分裂作用是由 175 kD 介导的 甘露糖识别受体（ASM-MR）。 具体目标是： 1) 确定 p42 和 p44 丝裂原激活蛋白激酶的贡献 (p42/44MAPK) 和富含甘露糖基的 70-kD 核糖体蛋白 S6 激酶 (p70S6k) 糖蛋白诱导气道平滑肌细胞（ASMC）有丝分裂。 2） 确定 p72Syk 和 p125 粘着斑激酶的贡献 (p125FAK) 在富含甘露糖基的糖蛋白诱导的 ASMC 有丝分裂中。 3) 至 研究气道平滑肌甘露糖受体（ASM-MR）和 整合素在富含甘露糖基的信号转导中合作 糖蛋白诱导的有丝分裂。 为了实现具体目标 1 和 2， 上述激酶的活性和响应 Hex 和 a 的细胞增殖 在存在和不存在特定的情况下富含甘露糖基的新糖蛋白 抑制剂、反义寡核苷酸、正义寡核苷酸和乱序寡核苷酸将 相关的。 实现具体目标3、免疫共沉淀、交叉 连锁分析、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 表达将被多次脉冲抑制 反义寡核苷酸及其作用将在富含甘露糖基的情况下进行检查 糖蛋白诱导的有丝分裂。 拟议的研究旨在 增进对气道平滑肌底层机制的了解 增殖。

项目成果

beta-Hexosaminidases: potent mitogens of airway smooth muscle.

β-氨基己糖苷酶：气道平滑肌的有效有丝分裂原。

• DOI: 10.2500/108854198778612654
• 发表时间: 1998
• 期刊: Allergy and asthma proceedings
• 影响因子: 2.8
• 作者: Lew,DB

Enzymic methylation of arginyl residues in -gly-arg-gly- peptides.

• DOI: 10.1042/0264-6021:3480573
• 发表时间: 2000-06
• 期刊: The Biochemical journal
• 作者: Y. Hyun;D. Lew;S. Park;Chan-Wha Kim;W. Paik;Sangduk Kim

Effect of dexamethasone on bovine airway smooth muscle cell proliferation.

地塞米松对牛气道平滑肌细胞增殖的影响。

• DOI: 10.3109/02770909909054558
• 发表时间: 1999
• 期刊: The Journal of asthma : official journal of the Association for the Care of Asthma
• 作者: Dixon,ER;Weinberg,JA;Lew,DB
• 通讯作者: Lew,DB