Carbonic anhydrase IX and pulmonary endothelial cell acidosis during infection

感染过程中碳酸酐酶 IX 与肺内皮细胞酸中毒

• 批准号: 10215318
• 负责人: Ji Young Lee
• 金额: $ 16.01万
• 依托单位: UNIVERSITY OF SOUTH ALABAMA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10215318
• 关键词: Acidosis; Acids; Advisory Committees; Affect; Alabama; Alveolar; Amyloid; Amyloid Proteins; Award; Bicarbonates; Biological Assay; Biological Markers; Biology; Blood; Blood capillaries; Bronchoalveolar Lavage Fluid; Buffers; Carbon Dioxide; Catalytic Domain; Cellular Metabolic Process; Cellular biology; Cleaved cell; Clinical; Communication; Critical Care; Critical Illness; Culture Media; Development; Development Plans; Diagnostic; Doctor of Philosophy; Endothelial Cells; Endothelium; Enzymes; Exhibits; Faculty; Family; Feedback; Functional disorder; Funding; Gases; Goals; Human; Impairment; In Vitro; Infection; Internal Medicine; K-Series Research Career Programs; Knowledge; Leadership; Lung; Lung infections; Medicine; Membrane; Mentors; Metabolic; Metabolic acidosis; Metabolism; Outcome; Patients; Perfusion; Physicians; Physiological; Physiology; Pneumonia; Production; Protein Isoforms; Proteoglycan; Protons; Pseudomonas aeruginosa; Pseudomonas aeruginosa infection; Pulmonary Edema; Rattus; Regulation; Research; Research Personnel; Resistance; Role; Sampling; Scientist; System; Testing; Therapeutic; Time; Training; Training Programs; Translating; Translational Research; Translations; Transmembrane Domain; Universities; Vocational Guidance; Water; Work; base; blood perfusion; carbonate dehydratase; career; career development; cell motility; cytotoxic; cytotoxicity; genetic approach; hexokinase; improved; in vivo; injury and repair; insight; lung injury; meetings; migration; mortality; mouse model; preservation; professor; pyruvate dehydrogenase; repaired; skills; therapeutic target

PROJECT SUMMARY This Mentored Clinical Scientist Research Career Development Award (K08) proposal describes a five-year training program to support Ji Young Lee, an MD/PhD pulmonologist/intensivist, for her research-intensive independent physician-scientist career. The primary mentor, Dr. Troy Stevens, a world leader in the field of lung biology with a track record of excellence in training junior faculty, and the advisory committee consisting of accomplished researchers with diverse expertise, will provide scientific feedback and career guidance throughout the award period. The proposed work will be carried out at the highly interdisciplinary and well-funded Center for Lung Biology, a strength of the University of South Alabama. The candidate was recently appointed as assistant professor of Physiology and Cell Biology and Internal Medicine on the tenure-accruing investigator track with 75% of protected research time. Twenty five percent time is allocated for clinical activities in the division of pulmonary and critical care medicine, where she is facilitating translational research. The career development plan lays out didactic coursework, scientific meetings and professional development opportunities that will help the candidate obtain core knowledge and scientific communication and academic leadership skills, and develop R01 funding. The proposed research plan focuses on improving our understanding of the effects of acidosis on pneumonia, and establishing the conceptual basis for diagnostic and therapeutic translation. Acidosis is common in critically ill pneumonia patients, and is associated with high mortality. The pathophysiology of acidosis in pneumonia is poorly understood, and current therapies fail to improve major outcomes. Our studies have shown that pulmonary microvascular endothelial cells (PMVECs) utilize the carbonic anhydrase IX (CA IX) isoform to regulate pH, metabolism and migration. We also demonstrated that Pseudomonas aeruginosa infection of PMVECs induces release of cytotoxic amyloid proteins, which disrupts the alveolar capillary membrane. These cytotoxic amyloids induce soluble CA IX shedding from PMVECs which compromises their repair potential. Based on these preliminary studies, we test the hypothesis that P. aeruginosa infection induces cytotoxic amyloid production that leads to shedding of soluble CA IX in PMVECs, increasing lung injury. Specific aims test the hypotheses that: 1) CA IX is critical to the acid regulation, metabolism and migration of PMVECs and pulmonary endothelial barrier integrity; and, 2) P. aeruginosa infection elicits cytotoxic amyloid production, causing CA IX shedding in PMVECs, which increases lung injury. In vitro, we will use genetic approaches and endothelial cell functional assays to evaluate the effects of acidosis and the role of specific CA IX functional domains during physiologic and infectious conditions. In vivo and ex vivo, we will use acidosis, pneumonia and isolated lung perfusion mouse models to translate in vitro findings. Successful completion of this study will provide new insights into the mechanisms underlying acidosis in pneumonia and help identify CA IX and cytotoxic amyloids as biomarkers and therapeutic targets for pneumonia.

项目摘要 这个指导临床科学家研究职业发展奖（K 08）提案描述了一个为期五年的 培训计划，以支持Ji Young Lee，MD/PhD肺病学家/重症监护医师，为她的研究密集型 独立的医生科学家生涯。主要导师，特洛伊史蒂文斯博士，在肺领域的世界领导者 生物学，在培训初级教师方面有着卓越的记录，咨询委员会由以下人员组成： 具有不同专业知识的有成就的研究人员将提供科学反馈和职业指导 整个颁奖期间。拟议的工作将在高度跨学科和资金充足的 肺生物学中心，南亚拉巴马大学的强项。候选人最近被任命为 作为生理学、细胞生物学和内科学的助理教授， 跟踪75%的受保护研究时间。百分之二十五的时间分配给该部门的临床活动 肺和重症监护医学，在那里她是促进转化研究。职业发展 该计划列出了教学课程，科学会议和专业发展机会，这将有助于 候选人获得核心知识和科学沟通和学术领导技能，并发展 R 01融资拟议的研究计划的重点是提高我们对酸中毒的影响的理解， 肺炎，并建立诊断和治疗翻译的概念基础。酸中毒很常见 在重症肺炎患者中，与高死亡率相关。酸中毒的病理生理学 对肺炎知之甚少，目前的治疗未能改善主要结果。我们的研究显示 肺微血管内皮细胞（PMVEC）利用碳酸酐酶IX（CA IX）同种型， 调节pH、代谢和迁移。我们还证明了铜绿假单胞菌感染 PMVEC诱导细胞毒性淀粉样蛋白的释放，其破坏肺泡毛细血管膜。这些 细胞毒性淀粉样蛋白诱导可溶性CA IX从PMVEC脱落，这损害了它们的修复潜力。 基于这些初步研究，我们检验了铜绿假单胞菌感染诱导细胞毒性淀粉样蛋白的假设 导致PMVEC中可溶性CA IX脱落，增加肺损伤。具体目标检验 假设：1）CA IX是至关重要的酸调节，代谢和迁移的PMVEC和肺 内皮屏障完整性; 2）铜绿假单胞菌感染诱发细胞毒性淀粉样蛋白产生，引起CA IX PMVEC脱落，增加肺损伤。在体外，我们将使用遗传方法和内皮细胞 功能测定，以评估酸中毒的影响和特定的CA IX功能结构域的作用， 生理和感染条件。在体内和体外，我们将使用酸中毒，肺炎和离体肺 灌注小鼠模型，以转化体外研究结果。成功完成这项研究将提供新的见解 研究肺炎中酸中毒的潜在机制，并帮助识别CA IX和细胞毒性淀粉样蛋白， 肺炎的生物标志物和治疗靶点。