Airway epithelial regulation of fibroblast hyaluronan and versican production in asthmatic airway inflammation

项目总结/摘要 该项目是NIH指导临床科学家研究职业发展奖（K 08）的应用程序博士。 斯蒂芬·里夫斯是宾夕法尼亚大学儿科系的代理助理教授， 华盛顿（UW）。Reeves博士已经完成了儿科肺部医学的临床培训， 在儿科哮喘的治疗中具有临床和研究兴趣。里夫斯博士的研究兴趣 是了解气道上皮细胞（AEC）在调节产生额外的- 细胞基质（ECM）的气道间充质细胞和改变，在这一过程中，可能倾向于 气道炎症他的长期职业目标是建立自己作为一个独立资助的原则 研究人员使用基础科学和转化方法在人类细胞中研究这些机制。 为了实现这一目标，Reeves博士请求NIH K 08支持额外的培训， 在以下特定领域的指导：（1）气道细胞生物学的额外培训，包括细胞-细胞和 细胞-基质相互作用;（2）ECM表征和定量方法学的额外培训;（3） 与基因敲除和基因编辑相关的实验室技术的额外培训;（4）额外培训 蛋白质组学方法和数据分析;（5）参加科学会议，职业发展 研讨会，以及与本项目相关的额外课堂培训;以及（6） 发展独立的研究重点，目标是向科学独立过渡。 在本申请中，Reeves博士提出了进一步研究AEC信号传导所起作用的研究。 在驱动HA和VCAN沉积在ECM中的HLF。这些研究将侧重于以下领域： 目的1）确定原发性哮喘AEC是否驱动HLF产生富含HA和VCAN的ECM， 与来自健康儿童的AEC相比;具体目的2）确定HA和VCAN富集的ECM是否 由哮喘AEC调节的HLF产生的白细胞粘附、迁移和活化增加， 与由与健康AEC共培养的HLF产生的ECM相比;具体目的3）确定是否临床上 AEC供体肺功能和气道炎症指标与HA和VCAN相关性研究 在ECM中的积累，以及由ECM产生的增强的白细胞粘附、迁移和活化， 共培养的HLF。这些研究有希望进一步了解AEC如何调节ECM HLF沉积，最终保护免受或易患炎症变化，这是在 目前知之甚少。此外，预计这些研究将提供出版物， 初步数据需要制定一个独立的研究方向，并在年底申请R 01资金 职业发展支持。

PROJECT SUMMARY / ABSTRACT This project is a NIH Mentored Clinical Scientist Research Career Development Award (K08) application for Dr. Stephen Reeves, an Acting Assistant Professor in the Department of Pediatrics at the University of Washington (UW). Dr. Reeves has completed his clinical training in Pediatric Pulmonary Medicine and has both a clinical and research interest in the treatment of pediatric asthma. Dr. Reeves’ specific research interest is understanding the role played by the airway epithelial cells (AECs) in regulating the production of extra- cellular matrix (ECM) by airway mesenchymal cells and alterations in this process that may predispose to airway inflammation. His long-term career goal is to establish himself as an independently-funded principle investigator studying these mechanisms in human cells using basic science and translational approaches. In order to achieve this goal, Dr. Reeves is requesting the NIH K08 support for additional training and mentorship in the following specific areas: (1) additional training in airway cell biology including cell-cell and cell-matrix interactions; (2) additional training in the methodology of ECM characterization and quantitation; (3) additional training in laboratory techniques related to gene knockdown and gene editing; (4) additional training in proteomic approaches and data analysis; (5) attendance of scientific conferences, career development seminars, and additional classroom-based training relevant to this project; and (6) mentorship in the development of an independent research focus with a goal of transitioning to scientific independence. In the present application, Dr. Reeves proposes studies to further investigate the role that AEC signaling plays in driving HA and VCAN deposition in the ECM by HLF. These studies will focus on following areas: Specific Aim 1) determining if primary asthmatic AECs drive HLFs to produce an ECM enriched in HA and VCAN as compared to AECs from healthy children; Specific Aim 2) determining if a HA- and VCAN-enriched ECM produced by HLFs conditioned by asthmatic AECs increases leukocyte adhesion, migration, and activation, as compared to ECM produced by HLFs co-cultured with healthy AECs; Specific Aim 3) determining if clinical indices of lung function and airway inflammation among AEC donors correlate with HA and VCAN accumulation in ECM, and with enhanced leukocyte adhesion, migration, and activation by ECM produced by co-cultured HLFs. These studies hold promise in furthering our understanding of how AECs may regulate ECM deposition by HLFs that ultimately protect against or predispose to inflammatory changes, which is at the present poorly understood. Furthermore, it is anticipated these studies would provide the publications and preliminary data needed to develop an independent research direction and apply for R01 funding at the end of the career development support.