Effect of chronic cigarette smoking on human microvascular endothelial cell metabolism and function

项目摘要/摘要 这是一份为期18个月的研究计划书，将允许申请者在 肺内科。申请人目前是美国国立卫生研究院的肺科学和重症监护医学研究员。 科罗拉多大学/国家犹太人健康中心。伊琳娜·彼得雷切博士，一位著名的内科科学家， 在肺气肿、肺损伤和修复、肺血管生物学和鞘脂信号转导方面的专业知识，将是 申请者的主要导师和赞助人。卡琳娜·瑟班博士，一位在肺先天方面有专长的内科科学家 免疫反应、单核细胞-内皮细胞相互作用和α1抗胰蛋白酶缺乏症将是共同的导师。 这项拟议的研究将调查长期吸烟(CS)损害健康的机制 人肺微血管内皮细胞(HLMVEC)承受适当的动态平衡和应激反应的能力 诱导自噬(一个生存和修复过程)并进行血管生成(损伤修复所必需的)。 这一机制具有重要的临床意义，因为HLMVEC是肺泡的重要组成部分 气体交换、宿主防御和损伤修复所需的膜。应用的科学前提 依赖于通过S1P受体1(S1P1)传递的S1P信号的关键作用来确保LMVEC 生存、增殖和屏障功能。来自主要赞助商的稳健的已发布和初步数据 实验室研究表明，增强S1P-S1P1信号通路可保护LMVECs免受不利影响 急性CS暴露。然而，慢性CS暴露对LMVEC存活和功能的影响尚未见报道 已经被定义了。申请人假设HLMVEC在慢性CS后发生适应不良变化 暴露，其特征是自噬受损，由于S1P1信号减弱而导致血管生成减弱， 这可以通过增强S1P-S1P1信号来逆转。申请者将分离并培养初级 HLMVEC来自有慢性吸烟史或终身不吸烟者的未鉴定的人类供体肺 并研究基线S1P-S1P1信号的差异及其修复反应的质量(自噬和 管形成，血管生成的替代物)到动态平衡和应激条件。最后，她会决定 这些功能依赖于完整的S1P-S1P1信号，通过酶和功能的获得和丧失 感受器。拟议的项目将是确定慢性疾病具体和新机制的重要一步 CS诱导的LMVEC损伤和功能障碍，因此可能为治疗方法的发展提供信息 继发于COPD的破坏性疾病，如肺气肿和肺动脉高压。 申请者将学习大量的翻译研究技能，并计划使用拟议的 该项目将成为她作为独立研究员未来研究拨款的基础，包括K08奖。 申请者得到了她的导师、她的研究咨询委员会和该部门的大力支持。

PROJECT SUMMARY/ABSTRACT This is an 18-month research proposal which will allow the applicant to develop an academic research career in Pulmonary Medicine. The applicant is currently a fellow in Pulmonary Sciences and Critical Care Medicine at the University of Colorado/National Jewish Health. Dr. Irina Petrache, an established physician-scientist with expertise in emphysema, lung injury and repair, lung vascular biology, and sphingolipid signaling, will be the applicant's primary mentor and sponsor. Dr. Karina Serban, a physician scientist with expertise in lung innate immune responses, monocyte-endothelial interactions, and alpha 1 antitrypsin deficiency, will be the co-mentor. The proposed research will investigate the mechanisms by which chronic cigarette smoking (CS) impairs the ability of human lung microvascular endothelial cells (HLMVEC) to undergo proper homeostatic and stress- induced autophagy (a survival and reparative process) and perform angiogenesis (required for injury repair). This mechanism has significant clinical implications, since HLMVEC are essential components of the alveolar membrane required for gas exchange, host defense, and injury repair. The scientific premise of the application relies on the key role of sphingosine-1 phosphate (S1P) signaling via S1P receptor 1 (S1P1) to ensure LMVEC survival, proliferation, and barrier function. Robust published and preliminary data from the primary sponsor's laboratory show that augmenting the S1P-S1P1 signaling pathway protects LMVECs from the detrimental effects of acute CS exposure. However, the effect of chronic CS exposure on LMVEC survival and function has not yet been defined. The applicant hypothesizes that HLMVEC develop maladaptive changes following chronic CS exposure, characterized by impaired autophagy and weakened angiogenesis due to diminished S1P1 signaling, which can be reversed by augmenting S1P-S1P1 signaling. The applicant will isolate and culture primary HLMVEC from de-identified human donor lungs with a history of chronic smoking or from lifelong nonsmokers and study differences in baseline S1P-S1P1 signaling and the quality of their repair responses (autophagy and tube formation, a surrogate of angiogenesis) to homeostatic and stress conditions. Finally, she will determine the dependency of these functions on intact S1P-S1P1 signaling by gain and loss of function of enzymes and receptors. The proposed project will be an important step in defining a specific and novel mechanism of chronic CS-induced LMVEC injury and dysfunction, and therefore may inform the development of therapies to treat devastating diseases such as emphysema and pulmonary hypertension secondary to COPD. The applicant will learn a large array of translational research skills and plans to use the results from the proposed project will form the basis of her future research grants as an independent researcher, including the K08 award. The applicant has strong support from her mentors, her research advisory committee, and the Division.