Using high T-bet-expressing cells and Serum Chemokines as Indicators of Disease Severity in Sarcoidosis

项目摘要/摘要 结节病是一种病因不明的全身性肉芽肿性疾病，累及多个器官，尤其是 肺部。它具有广泛的临床结果，包括进行性肺部疾病和多器官疾病。 参与其中。由于这些结果缺乏可靠或有效的标记物，结节病表现出许多 对临床医生的挑战，特别是考虑到用作治疗的免疫抑制疗法的毒性。 我的这个K23奖项的目标是确定特定的免疫细胞和蛋白质如何与系统器官相关 结节病患者的负担和肺功能。在实现这一目标的同时，我将追求严谨的 职业发展计划，扩大我在临床研究方法和T细胞生物学方面的技能。血液标记物 我将研究它们在生物学上与干扰素-γ有关，这是一种在结节病炎症中必不可少的细胞因子。这些标记物 包括：1)能产生大量干扰素-γ的特定辅助性T细胞亚群；2)血清中 干扰素-γ诱导的趋化因子将免疫细胞输送到炎症组织。我已经生成了初步数据 这些标记使用了由我的主要导师Dr。 劳拉·科思。感兴趣的T细胞群体由转录因子T-bet的产生来定义，该转录因子 调节干扰素-γ的产生。使用多参数流式细胞术，我鉴定了一组辅助T细胞 有明显高水平的T-bet蛋白(T-betHi细胞)。这些T-BetHi细胞几乎只存在于 肺功能测试(PFT)的受试者下降。对这些细胞的初步体外分析表明，它们有 产生干扰素-γ的能力非常高。目标1将确定在这些T-BetHi细胞中如何产生干扰素-γ 与它们的潜在致病性有关。第二组标志物包括三种干扰素-γ诱导的趋化因子 (CXCL9、CXCL10和CXCL11)，这是我最近发表的两篇文章的主题。我给你看了那份血清 这些趋化因子的水平与纵向肺功能变化和器官有明显的相关性 参与其中。这些关联可能是由于其潜在的生物学特性，因此具有特定的相关性 在疾病中。因此，在目标2和3中，我将确定T-BetHi细胞和这些趋化因子的关系 在两个结节病队列中的结果。我一直在帮助科思博士将受试者招募到这些队列中，我运行 研究访问，以收集我将在本提案中使用的临床数据。这些队列包括有新的 确诊的疾病，反复随访长达两年以上。我将确定这些标记是如何关联的 纵向为受影响的器官数目(目标2)和肺功能(目标3)。除了提供 科思博士直接参与了这些队列，她将分享她的实验室空间，并为测量这些数据提供支持 血液标记物。我还将受益于我的同事们的临床研究、生物统计学和专业知识， 伍德拉夫博士和艾伦博士。作为我的培训计划的一部分，我将补充我在攻读硕士期间学到的技能 通过额外的流行病学和统计学课程学习临床研究。在这个奖项期间，我将产生 申请R01基金成为独立资助的临床和翻译研究人员的数据。

PROJECT SUMMARY/ABSTRACT Sarcoidosis is a systemic granulomatous disease of unknown etiology that affects multiple organs, especially the lungs. It has a wide range of clinical outcomes, including progressive lung disease and multi-organ involvement. Due to the lack of reliable or validated markers for these outcomes, sarcoidosis presents many challenges to clinicians, especially in light of the toxicities from immunosuppressive therapies used as treatment. My goal for this K23 award is to determine how specific immune cells and proteins relate to systemic organ burden and pulmonary function in sarcoidosis patients. While accomplishing this goal, I will pursue a rigorous career development plan to expand my skills in clinical research methods and T cell biology. The blood markers I will study are biologically linked to IFN-γ, an essential cytokine in sarcoidosis inflammation. These markers include: 1) a specific subset of T helper cells that can produce high amounts of IFN-γ and 2) serum levels of chemokines induced by IFN-γ that traffic immune cells to inflamed tissues. I have generated preliminary data for these markers using a well-phenotyped longitudinal sarcoidosis cohort established by my primary mentor, Dr. Laura Koth. The T cell population of interest is defined by production of the transcription factor T-bet, which regulates IFN-γ production. Using multi-parameter flow cytometry, I identified a population of T helper cells that have distinctly high levels of T-bet protein (T-betHi cells). These T-betHi cells were present almost exclusively in subjects with pulmonary function test (PFT) declines. Initial in vitro assays of these cells suggest that they have exceptionally high capacity to produce IFN-γ. Aim 1 will determine how IFN-γ production in these T-betHi cells relates to their potential pathogenicity. The second group of markers includes three IFN-γ-induced chemokines (CXCL9, CXCL10, and CXCL11), which were the subject of two of my recent publications. I showed that serum levels of these chemokines had distinct associations with the longitudinal pulmonary function changes and organ involvement. These associations may be due to their underlying biology and therefore carry specific relevance in the disease. Thus, in Aims 2 and 3, I will determine the relationship of both T-betHi cells and these chemokines to outcomes in two sarcoidosis cohorts. I have been helping Dr. Koth enroll subjects into these cohorts and I run study visits to collect the clinical data I will use in this proposal. These cohorts include subjects with newly diagnosed disease who have repeated follow-up over up to two years. I will determine how these markers relate longitudinally to the number of organs affected (Aim 2) and pulmonary function (Aim 3). In addition to offering direct involvement with these cohorts, Dr. Koth, will share her lab space and provide support to measure these blood markers. I will also benefit from the clinical research and biostatistical and expertise of my co-mentors, Drs. Woodruff and Allen. As part of my training plan, I will supplement skills I have acquired during my Master's in Clinical Research through additional epidemiology and statistics coursework. During this award I will generate data to apply for R01 funding to become an independently funded clinical and translational researcher.