Enhancing Corticosteroid Sensitivity in Neonatal and Pediatric Lung Disease

项目总结/摘要 申请人的目标是发展必要的技能，成为独立的翻译 新生儿/儿科哮喘领域的研究人员。在生命早期发展的儿童哮喘是一种主要的医疗保健 负担皮质类固醇（CS）用于治疗，但与成人相比，儿童需要更高的CS剂量， 而一些发展为CS抵抗，气道重塑更大，难以治疗。 发育中气道的CS不敏感性在很大程度上是未知的。气道平滑肌（ASM）是呼吸道疾病的关键细胞类型。 哮喘，并表现出高反应性（AHR）、增殖和对炎症的反应性重塑。有 目前很少有关于ASM的发展如何导致新生儿/儿科哮喘或CS抵抗的信息。 总的假设是Th 1炎症（TNFα，IFNγ）通过以下途径诱导发育中气道的CS不敏感性： 破坏糖皮质激素受体表达和信号传导，导致ASM增殖和重塑增强。 在潜在的治疗方面，越来越多的证据表明维生素D（VitD）可增强CS敏感性，但很少有证据表明维生素D可增强CS敏感性。 没有关于基本机制的信息，特别是在发展气道方面（本建议的第二个重点）。经由3 目的是，申请人将逐步建立对研究CS不敏感性的研究独立性， 新生儿/儿童哮喘：目的1（K99阶段）：使用Th 1诱导的CS不敏感性体外模型，确定 炎症抑制GR信号传导和活性的机制。目标2（K99/R 00 阶段）：使用Th 1诱导的CS不敏感性的体外模型，确定VitD增强CS不敏感性的机制。 GR信号在人类ASM发育中的作用特定目标3（R 00阶段）：在新型CS不敏感与CS敏感中 新生小鼠过敏性气道炎症模型，确定CS和骨化三醇之间的相互作用， 缓解AHR和重塑。指导阶段将研究Th 1细胞因子如何破坏CS信号传导， 人胎儿ASM（妊娠18-22周）。申请人将接受研究细胞，分子和 通过新生儿CS不敏感性的体外模型与糖皮质激素受体信号传导相关的表观遗传机制 包括Th 1细胞因子。补充教学，智力和专业培训将有助于准备 R 00阶段的申请人，他将研究与VitD如何增强CS敏感性有关的机制， 使用CS不敏感性的体外和体内新生小鼠模型开发ASM。总之，这些小说 研究将加强目前对生命早期炎症如何破坏糖皮质激素受体的理解 信号转导，并将确定VitD改善CS敏感性的潜力。申请人将由资深， 在ASM生理学、肺免疫学、糖皮质激素信号传导、 和哮喘。重要的是，该项目将为申请人建立独立的研究提供基础 新生儿/儿科气道疾病项目。

PROJECT SUMMARY/ABSTRACT The applicant's goals are to develop the necessary skills to become an independent translational researcher in the area of neonatal/pediatric asthma. Childhood asthma developing early in life is a major healthcare burden. Corticosteroids (CS) are used therapeutically, but compared to adults, children require higher CS doses, and some develop CS resistance with greater airway remodeling that is difficult to treat. The mechanisms underlying CS insensitivity in developing airway are largely unknown. Airway smooth muscle (ASM) is a key cell type in asthma, and exhibits hyperreactivity (AHR), proliferation, and remodeling in response to inflammation. There is currently little information on how developing ASM contributes to neonatal/pediatric asthma, or to CS resistance. The overall hypothesis is that Th1 inflammation (TNFα, IFNγ) induces CS insensitivity in developing airway by disrupting glucocorticoid receptor expression and signaling, leading to enhanced ASM proliferation and remodeling. In terms of potential therapy, there is increasing evidence for Vitamin D (VitD) enhancing CS sensitivity, but little to no information on underlying mechanisms, particularly in developing airway (a second focus of this proposal). Via 3 Aims, the applicant will progressively build towards research independence investigating CS insensitivity in neonatal/pediatric asthma: Aim 1 (K99 Phase): Using an in vitro model of Th1 induced CS insensitivity, determine mechanisms by which inflammation inhibits GR signaling and activity in developing human ASM. Aim 2 (K99/R00 Phases): Using an in vitro model of Th1 induced CS insensitivity determine mechanisms by which VitD enhances GR signaling in developing human ASM. Specific Aim 3 (R00 Phase): In novel CS-insensitive vs. –sensitive newborn mouse models of allergic airway inflammation, determine interactions between CS and calcitriol in alleviating AHR and remodeling. The mentored phase will examine how Th1 cytokines disrupt CS signaling in human fetal ASM (18-22 week gestation). The applicant will receive training in investigating cellular, molecular, and epigenetic mechanisms related to glucocorticoid receptor signaling via an in vitro model of neonatal CS insensitivity involving Th1 cytokines. Complementary didactic, intellectual, and professional training will help prepare the applicant for the R00 phase where he will examine mechanisms relating to how VitD may enhance CS sensitivity in developing ASM using in vitro and novel in vivo neonatal mouse models of CS insensitivity. Together, these novel studies will enhance current understanding of how inflammation early in life disrupts glucocorticoid receptor signaling, and will identify the potential for VitD to improve CS sensitivity. The applicant will be mentored by senior, established investigators with substantial expertise in ASM physiology, lung immunology, glucocorticoid signaling, and asthma. Importantly, this project will provide a foundation for the applicant to establish an independent research program in neonatal/pediatric airway disease.