Mechanisms of gland proliferation in COPD

COPD 腺体增殖机制

• 批准号: 7114302
• 负责人: Steven Richard HAYS
• 金额: $ 14.88万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7114302
• 关键词: acinar cell; bronchoscopy; cell differentiation; cell proliferation; chronic obstructive pulmonary disease; clinical research; exocrine glands; growth factor receptors; histogenesis; histology; human subject; immunocytochemistry; laser capture microdissection; mucins; mucosa; mucus; patient oriented research; respiratory epithelium; secretion; smoking; tobacco abuse

DESCRIPTION (provided by applicant): This K23 application proposes a program of patient-oriented research training and studies that will apply novel techniques to investigate the mechanisms of gland proliferation in COPD. The longterm objective is to address persistent gaps in knowledge that prevent the development of novel and specific treatments of gland proliferation in COPD. The proposed approach will apply recently developed techniques for morphometry (design-based stereology), cell isolation (laser capture microdissection), gene expression analysis (two-step real time PCR) and a tissue culture model of gland development. Application of these techniques to bronchial biopsy specimens obtained by bronchoscopy will make it possible to study gene expression in submucosal glands isolated from well characterized human subjects. The specific aims of the proposal are to 1) To determine if submucosal gland proliferation occurs in non-fatal COPD and is associated with serous cell transdifferentiation and MUC5B overexpression. 2)To determine if submucosal gland proliferation in COPD occurs because of new gland growth from the surface epithelium. 3)To determine the role of EGFR and TGFalpha as inducers of submucosal gland proliferation in COPD. We propose two studies: 1) a cross sectional morphologic analysis of airway biopsies from smokers and healthy controls. Using design based stereology we will compare the volume of submucosal glands, the ratio of serous cells to mucous cells in glands, and the volume of MUC5B, TGFalpha and EGFR staining in the glands of smoker's compared to healthy controls. In addition we will quantify the number of terminal collecting ducts per surface area of basal lamina to determine if new gland growth occurs in smokers. 2) an experiment using bronchial biopsies in culture as a model for gland development. Using this model, in combination with morphometry, laser capture microdissection and RT-PCR, we will study the effects of TGFalpha and EGFR on the development of tubuloacinar structures in bronchial biopsy explants. The candidate will complement these studies with a curriculum of advanced training in molecular biology methods of clinical research.

描述（由申请人提供）： 该 K23 申请提出了一项以患者为导向的研究培训和研究计划，该计划将应用新技术来研究 COPD 中腺体增殖的机制。长期目标是解决长期存在的知识空白，这些空白阻碍了慢性阻塞性肺病腺体增殖的新型和特异性治疗方法的开发。所提出的方法将应用最近开发的形态测量技术（基于设计的体视学）、细胞分离（激光捕获显微切割）、基因表达分析（两步实时 PCR）和腺体发育的组织培养模型。将这些技术应用于通过支气管镜检查获得的支气管活检标本将使研究从特征明确的人类受试者中分离的粘膜下腺体中的基因表达成为可能。该提案的具体目的是 1) 确定粘膜下腺增殖是否发生在非致命性 COPD 中，并且与浆液细胞转分化和 MUC5B 过度表达相关。 2)确定COPD中粘膜下腺体增殖是否是由于表面上皮生长出新的腺体所致。 3)确定EGFR和TGFα作为COPD粘膜下腺增殖诱导剂的作用。我们提出两项研究：1）对吸烟者和健康对照者的气道活检进行横断面形态学分析。使用基于设计的体视学，我们将比较吸烟者与健康对照者的粘膜下腺体的体积、腺体中浆液细胞与粘液细胞的比率以及粘膜下腺体中MUC5B、TGFα和EGFR染色的体积。此外，我们将量化基底层每表面积的终末集合管的数量，以确定吸烟者是否出现新的腺体生长。 2）使用培养中的支气管活检作为腺体发育模型的实验。使用该模型，结合形态测量、激光捕获显微切割和 RT-PCR，我们将研究 TGFα 和 EGFR 对支气管活检外植体中管状腺泡结构发育的影响。候选人将通过临床研究分子生物学方法的高级培训课程来补充这些研究。