Enhancing Corticosteroid Sensitivity in Neonatal and Pediatric Lung Disease

项目摘要/摘要 申请者的目标是发展成为独立翻译所需的技能 新生儿/儿童哮喘领域的研究人员。儿童哮喘在生命早期发展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， 一些患者出现CS抵抗，并伴有难以治疗的更大的气道重塑。潜在的机制 CS在发展中的呼吸道不敏感在很大程度上是未知的。气道平滑肌(ASM)是一种重要的细胞类型。 哮喘，并对炎症反应表现出高反应性(AHR)、增殖和重塑。的确有 目前，关于发育中的ASM如何促进新生儿/儿童哮喘或CS抵抗的信息很少。 总体假设是Th1型炎症(肿瘤坏死因子α、干扰素γ)通过以下途径诱导CS在发育中的气道不敏感 破坏糖皮质激素受体的表达和信号，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)可以增强CS的敏感性，但对 没有关于潜在机制的信息，特别是在发展呼吸道方面(本提案的第二个重点)。通过3 目的，申请人将逐步建立研究独立调查政务司司长不敏感在 新生儿/儿童哮喘：目标1(K99期)：使用Th1诱导的CS不敏感的体外模型，确定 炎症抑制人ASM发育中GR信号和活性的机制。目标2(K99/R00 阶段)：使用Th1诱导的CS不敏感的体外模型确定VitD增强的机制 GR信号在人类ASM发育中的作用特定目标3(R00阶段)：在新的CS不敏感与敏感中 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Th1细胞因子如何扰乱CS信号。 人胎儿自闭症(妊娠18-22周)。申请者将接受研究细胞、分子和 通过新生儿CS不敏感体外模型与糖皮质激素受体信号相关的表观遗传学机制 涉及Th1细胞因子。辅助性的教学、智力和专业培训将有助于为 R00阶段的申请人将在该阶段检查与VitD如何提高CS敏感性有关的机制 用体外和体内新的CS不敏感新生小鼠模型开发ASM。总而言之，这些小说 研究将加强目前对生命早期炎症如何扰乱糖皮质激素受体的理解 并将确定VitD改善CS敏感性的潜力。申请者将由前辈、 在ASM生理学、肺免疫学、糖皮质激素信号转导方面拥有丰富专业知识的资深研究人员， 还有哮喘。重要的是，这个项目将为申请者建立独立的研究提供基础 新生儿/儿童呼吸道疾病方案。

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.