The Role of 2-Pore Domain Potassium Channels in Acute Lung Injury.

描述（由申请人提供）：尽管在肺病学和重症监护医学方面取得了进展，但急性肺损伤（ALI）患者的死亡率仍然很高，这可能是因为对这种疾病的潜在机制知之甚少。ALI患者通常需要高氧浓度和正压通气，尽管这两种治疗都会加重持续性肺损伤。虽然高氧激活的信号级联是众所周知的，但肺泡扩张后的“机械转导”却知之甚少。最近的文献表明，2孔结构域钾（K2 P）通道可能作为机械传感器和机械转换器，并参与刺激-分泌偶联。然而，对K2 P通道在肺中的表达和潜在功能知之甚少。我们提出了一个大胆而新颖的假设，即K2 P通道在肺上皮细胞中表达，并且由高氧、机械拉伸和TNF-α暴露（类似于ALI中遇到的环境）引起的致病性K2 P通道调节导致上皮细胞的炎症介质分泌失调和上皮屏障功能丧失，这是ALI的两个标志。我们的首要目标是研究导致高氧和机械牵张诱导的肺损伤的机制，并确定K2 P通道作为寻找针对ALI的创新治疗策略的新靶点。具体而言，我们将使用体外和体内方法，包括K2 P敲除小鼠[1]，使用分子技术、免疫组织化学和膜片钳研究，研究高氧、机械拉伸和TNF-α对培养的小鼠和原代大鼠和人肺泡上皮细胞中K2 P通道表达和功能的影响，[2]为了确定K2 P通道在培养的和原代肺泡上皮细胞的炎症介质分泌中的作用，以及在K2 P敲除小鼠的支气管肺泡灌洗液中的作用，和[3]证明K2 P通道通过Ca 2+依赖性紧密连接磷酸化调节上皮屏障功能。 我们拥有独特的专业知识和技术能力来研究高氧和机械拉伸在体外和体内ALI模型中的作用。此外，Jaggar博士拥有独特的设置来测量整体和局部细胞内Ca 2+浓度。UTHSC的学术环境，优秀的导师，丰富的合作机会，以及对我的研究成功的机构，部门和部门的承诺，提供了智力基础设施和财政支持，以保证我在5年内向独立研究资金的进展，包括R 01奖。这些长期目标将通过每年至少在国际会议上发表2篇论文和2篇摘要演讲来实现，并辅以正式课程，以及沃茨博士，阿南德博士和我的职业咨询委员会的密切指导。

DESCRIPTION (provided by applicant): Despite advances in Pulmonology and Intensive Care Medicine, mortality rates of patients with Acute Lung Injury (ALI) remain high, perhaps because the underlying mechanisms of this disease are poorly understood. Patients with ALI routinely require high oxygen concentrations and positive-pressure ventilation, although both therapies accentuate ongoing lung injury. While the signaling cascades activated by hyperoxia are well known, "mechano-transduction" after alveolar distension is poorly understood. Recent literature suggests that 2-pore domain potassium (K2P) channels may act as mechano-sensors and mechano-transducers, and participate in stimulus-secretion coupling. However, little is known about the expression and potential functions of K2P channels in the lung. We propose a bold and novel hypothesis that K2P channels are expressed in lung epithelial cells, and that pathogenic K2P channel regulation caused by hyperoxia, mechanical stretch and TNF-a exposure, an environment similar to the one encountered in ALI, results in dysregulation of inflammatory mediator secretion from epithelial cells, and in loss of epithelial barrier function, two hallmarks of ALI. Our overarching objective is to investigate the mechanisms leading to hyperoxia- and mechanical stretch- induced lung injury, and to identify K2P channels as a new target in the search for innovative therapeutic strategies against ALI. Specifically, we will use both in vitro and in vivo approaches including K2P knockout mice to [1] investigate the effects of hyperoxia, mechanical stretch and TNF-a on K2P channel expression and function in cultured mouse and primary rat and human alveolar epithelial cells using molecular techniques, immunohistochemistry and patch clamp studies, [2] to determine the role of K2P channels in inflammatory mediator secretion from cultured and primary alveolar epithelial cells, and in broncho-alveolar lavage fluid from K2P knockout mice, and [3] to demonstrate that K2P channels regulate epithelial barrier function via Ca2+-dependent tight junction phosphorylation. We have the unique expertise and technical capabilities to study the effects of hyperoxia and mechanical stretch in both in vitro and in vivo models of ALI. In addition, Dr. Jaggar has an inimitable setup to measure global and localized intracellular Ca2+ concentrations. The academic environment at UTHSC, the outstanding mentorship, rich opportunities for collaborations, and the institutional, departmental, and divisional commitment to my research success provide the intellectual infrastructure and the financial support to guarantee my progress towards independent research funding, including an R01 award, within 5 years. These long-term goals will be achieved by targeting a minimum of 2 publications and 2 abstract presentations per year at international meetings, supplemented by formal coursework, and the close mentorship of Dr. Waters, Dr. Anand and my Career Advisory Committee.