EGF AND TGFA IN DEVELOPING AIRWAY EPITHELIUM

EGF 和 TGFA 在气道上皮发育中的作用

• 批准号: 3353690
• 负责人: MILDRED T STAHLMAN
• 金额: $ 15.14万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-09-30 至 1990-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3353690
• 关键词: developmental genetics; electron microscopy; epidermal growth factor; gene expression; genetic transcription; growth factor receptors; histochemistry /cytochemistry; histogenesis; human fetus tissue; human tissue; immunochemistry; immunocytochemistry; in situ hybridization; infant human (0-1 year); lung; messenger RNA; peptide hormone biosynthesis; postmortem; respiratory distress syndrome of newborn; respiratory epithelium; transforming growth factors

Our objective is to determine if growth factors can be related to the physiology of intraparenchymal airway development and response to injury in the human lung. In the future, new biologial strategies for the treatment of newborn lung disease may involve the use of growth factors. Precise information about the synthesis of growth factors and the availability of their receptors may contribute to the design of such therapies. The emphasis of this project is to define the cellular sites of synthesis and changes in the synthesis of epidermal growth factor (EGF), transforming growth factor alpha (TGFalpha) and the EGF receptor (EGF-R) during lung development and the perturbed response to injury seen in bronchopulmonary dysplasia (BPD). Three major hypotheses will be tested: (1) select cell populations of the intraparenchymal conducting airways produce EGF, TGFalpha and EGF-R; (2) EGF, or a similar ligand for EGF-R, exerts autocrine or paracrine effects contributing to the development (and repair) of pulmonary epithelium; (3) changes in the production of EGF, or growth factor ligands for EGF-R, may occur during lung development and BPD. These hypotheses will be tested with the following specific aims. (1) Changes in the morphological differentiation of airway epithelium during lung development will be defined by light and electron microscopy. (2) Sensitive immunocytochemistry (IC) will be used to identify the specific cell types containing immunoreactive EGF, TGFalpha and EGF-R. Since neuroendocrine cells may uptake and store peptide hormones and the EGF: receptor complex is rapidly internalized and degraded after EGF binding, IC alone does not establish the sites of synthesis (or lack of synthesis) of these products. (3) The more direct method of nucleic acid in situ hybridization histochemistry (IHH) will be used to identify the cellular sites of transcription of EGF, TGFalpha and EGF-R mRNA. IHH alone does not establish that the target mRNA is translated and processed to the peptide. Hence, identification of both the mRNA for a peptide and the immunoreactive peptide in the same or morphologically identical (and phenotypically defined) cell increases the probability that this cell is synthesizing the peptide product. (4) Since EGF may be involved in the repair response, the sites of synthesis of EGF, TGFalpha and EGF-R will be identified in BPD lungs. Growth factor gene expression may be under developmental control and/or altered during the response to injury, therefore (5) relative changes in the gene expression of EGF, TGFalpha and EGF-R will be defined by quantitative hybridization studies of lung during normal development and BPD. (6) Additional studies will identify the sites of and changes in the synthesis of other promising growth-related peptides. These studies will determine the in vivo sites of EGF synthesis and provide in vivo evidence of a physiological role for EGF during lung development and the response to injury found in BPD.

我们的目标是确定生长因子是否与 肺实质内气道发育和对损伤反应的生理学 人类的肺 在未来，新的生物治疗策略 新生儿肺部疾病可能涉及使用生长因子。 精确 关于生长因子的合成和 它们的受体可能有助于这种疗法的设计。 的 该项目的重点是确定合成的细胞位点， 表皮生长因子（EGF）的合成变化，转化 生长因子α（TGF α）和EGF受体（EGF-R）在肺 支气管肺炎的发展和对损伤的干扰反应 发育不良（BPD）。 三个主要假设将被测试：（1）选择细胞 实质内传导气道的群体产生EGF， TGF α和EGF-R;（2）EGF或EGF-R的类似配体， 促进发育（和修复）的自分泌或旁分泌效应 肺上皮细胞;（3）EGF的产生或生长的变化 EGF-R的因子配体可能发生在肺发育和BPD期间。 这些 将按照以下具体目标对假设进行检验。 (1)变化 肺发育过程中气道上皮的形态分化 将通过光学和电子显微镜来确定显影。 （二） 将使用灵敏的免疫细胞化学（IC）来鉴定特异性 含有免疫反应性EGF、TGF α和EGF-R的细胞类型。 以来 神经内分泌细胞可以摄取和储存肽激素和EGF： 受体复合物在EGF结合后迅速内化和降解，IC 单独并不能建立合成（或缺乏合成）的位点， 这些产品 (3)更直接的原位核酸法 杂交组织化学（IHH）将用于鉴定细胞 EGF、TGF α和EGF-R mRNA的转录位点。 IHH本身并不 确定靶mRNA被翻译并加工成肽。 因此，鉴定肽的mRNA和免疫反应性蛋白质两者， 肽在相同或形态上相同（和表型上 定义的）细胞增加了该细胞合成 肽产品。 (4)由于EGF可能参与修复反应， 将在BPD中鉴定EGF、TGF α和EGF-R的合成位点 肺 生长因子基因表达可能受发育控制 和/或在对损伤的反应期间改变，因此（5）相对 将确定EGF、TGF α和EGF-R基因表达的变化 通过正常发育期间肺的定量杂交研究， 波士顿警局 (6)进一步的研究将确定的网站和变化， 其他有前途的生长相关肽的合成。 这些研究将 确定EGF合成的体内位点，并提供体内证据 EGF在肺发育过程中的生理作用以及对 在BPD发现的伤口。