Defining the role of tuft cells in allergic airway disease

项目摘要/摘要 这份提案描述了一项实现候选人成为独立医生目标的5年计划- 研究呼吸道上皮和2型免疫系统之间相互作用的科学家。使用一个 在熟练的科学咨询和指导下，制定全面的课程和动手培训计划 在加州大学旧金山分校卓越的智力环境中，科塔斯博士将利用成熟的、 从导师的实验室获得专门的工具和技术，并新近精通初级呼吸道上皮 文化，使用人体组织的翻译研究，以及最先进的转录技术。到年底的时候 在这个项目中，她将拥有一个独特的工具集，使她有别于她的导师，并将拥有足够的 数据发起一个独立的研究小组，同时为科学界贡献新的知识。 这项建议的研究目的是阐明簇状细胞-特化的上皮细胞如何 了解的功能--在慢性2型呼吸道期间对邻近上皮细胞和免疫细胞行为的影响 发炎。由于慢性过敏性炎症是数百万人发病和死亡的根本原因 患有鼻息肉和2型高度哮喘等疾病的患者，对细胞和 设计新的治疗方法需要促成这些病理的分子。在初步数据中，科塔斯博士 发现慢性过敏性上呼吸道环境中的簇状细胞具有独特的“过敏性”表型 其特征是前列腺素E2(PGE2)的产生增加。这与转录的 PGE2在邻近上皮细胞上的信号，而在体外，PGE2刺激上皮液分泌。 在2型患者的支气管中也观察到了过敏丛状细胞的转录特征和PGE2的激活 高度哮喘，表明整个过敏性炎症的呼吸道都有类似的病理。这些 这一发现促使对过敏性呼吸道疾病的丛生细胞和丛生细胞衍生的PGE2进行进一步的检查。 在这项提案中，科塔斯博士将以她的初步数据为基础，探索丛生细胞和前列腺素E_2对 呼吸道动态平衡。Aim 1将利用小鼠体外细胞和体内全动物模型来检测丛生细胞- 黏液纤毛运动和呼吸道表面液体成分的依赖性改变。Aim 2将研究 丛状细胞和前列腺素E_2对过敏性呼吸道疾病小鼠模型2型免疫系统的影响。和 Aim 3将把焦点返回到人类受试者身上，并确定患者体内簇状细胞的表型和激活状态 2型高哮喘患者的下呼吸道。总之，拟议中的实验将改善我们的基础 了解簇状细胞如何促进过敏性呼吸道疾病，并可能鉴定新的分子和细胞 未来治疗的目标。

Project Summary/Abstract This proposal describes a 5-year plan to achieve the candidate’s goal of becoming an independent physician- scientist studying interactions between the airway epithelium and the type 2 immune system. With a comprehensive plan of coursework and hands-on training, guided by a skilled scientific advisory and mentoring team, and situated within the exceptional intellectual environment at UCSF, Dr. Kotas will utilize established, specialized tools and techniques from her mentor’s lab and become newly proficient in primary airway epithelial culture, translational studies using human tissues, and state-of-the-art transcriptional techniques. By the end of this project, she will possess a unique toolset that will differentiate her from her mentor and will have sufficient data to launch an independent research group while contributing new knowledge to the scientific community. The investigative purpose of this proposal is to elucidate how tuft cells—specialized epithelial cells with poorly understood function—influence the behavior of nearby epithelial and immune cells during chronic type 2 airway inflammation. As chronic allergic inflammation is the underlying cause of morbidity and mortality in millions of patients with diseases such as nasal polyps and type 2 high asthma, an improved understanding of the cells and molecules that contribute to these pathologies is needed to design new therapies. In preliminary data, Dr. Kotas finds that tuft cells in the chronically allergic upper airway environment adopt a distinct “allergic” phenotype characterized by increased production of prostaglandin E2 (PGE2). This is concurrent with a transcriptional signature of PGE2 on the neighboring epithelium, while in vitro, PGE2 stimulates epithelial fluid secretion. Transcriptional signatures of allergic tuft cells and PGE2 activation are also observed in the bronchus in type 2 high asthmatics, suggesting similar pathology throughout the allergically-inflamed respiratory tract. These findings urge further examination of tuft cells and tuft cell-derived PGE2 in allergic airway disease. In this proposal, Dr. Kotas will build upon her preliminary data to probe the effects of tuft cells and PGE2 on airway homeostasis. Aim 1 will use mouse cells in vitro and whole animal modeling in vivo to examine tuft cell- dependent alterations mucociliary movement and airway surface liquid composition. Aim 2 will examine the effects of tuft cells and PGE2 on the type 2 immune system in mouse models of allergic airway disease. And Aim 3 will return focus to human subjects and determine the phenotype and activation state of tuft cells in the lower airway in type 2 high asthma. Together, the proposed experiments will improve our fundamental understanding of how tuft cells contribute to allergic airway disease, and may identify new molecular and cellular targets for future therapy.