Targeted therapeutic modulation of inflammatory cytokines through manipulation of noncoding RNA regulation of innate immunity in atopic dermatitis

特应性皮炎(AD)是全球最常见的慢性炎症性皮肤病， 是由终末角质形成细胞分化缺陷和强大的2型免疫共同驱动的 回应。是什么控制了AD疾病的活动？为什么它对治疗的反应与 病人对病人？这些都是知之甚少的问题，并提出了大量未得到满足的需求 既有效又安全的疗法。在这个项目中，我们试图提供一个分子句柄 解释这些基本问题中的一些，通过解决潜在的未被探索的问题 以表观遗传机制为代表的免疫调节来源。我们的范式转变 假说的中心是关于免疫反应的表观遗传调节。 AD的发病机制，以及在一类新发现的长非编码RNA(LncRNAs)上， 免疫基因启动的lncRNAs(IPLs)，它利用特定的预先形成的染色质拓扑结构 细胞因子核室，以促进其表观遗传的启动和激活。我们建议 使用直到现在才使这一点成为现实的技术进步来测试和扩展我们的假设 有可能。我在这里概述了一个追求这个机会的计划，有三个具体目标。在目标1中(R61 阶段)，我们将建立协调表观遗传状态的上游分子事件 炎症基因。我们假设IPL是直接与 三维染色质结构可启动趋化因子启动子并增强亲核表达。 先天免疫细胞激活中的炎症反应。我们将描述这些机制的特征 IFL通过其调节IL-4和IL-13，Keystone Interleuins对诱导和 第二型反应在AD中的永久化。在目标2中，我们将鉴定新的蛋白质成分 直接与IPL相互作用以调节染色质景观的物质。我们还将表演 跨IL-4/IL-13基因座捕获染色体构象以确定是否改变 IPL-IL4/13的水平改变了整个基因座的染色体环路。最后，在目标3(R33阶段) 我们将在已建立的类似AD的小鼠模型中测试特异性靶向IPL-4/13的抑制剂 重述人类公元前十年。还将评估IPL抑制剂对感官的影响。 已知的影响AD行为的反应，以调节2型神经元的直接启动 细胞因子在AD的发病机制中起着关键作用。综合起来，我们的数据将 直接确定IPLS和三维染色质结构如何在合作中发挥作用 以减少基因固有噪音的方式，并允许强大的先天免疫基因激活。一个 通过直接操纵IPL活性更精确地微调趋化因子转录 代表了一种非常有价值的治疗策略，以实现AD的定制免疫调节。

Atopic Dermatitis (AD), the most common chronic inflammatory skin disorder worldwide, is driven by both terminal keratinocyte differentiation defects and strong type 2 immune responses. What controls AD disease activity? Why is its response to therapy different from patient to patient? These are poorly understood problems and presents a large unmet need for both effective and safe therapeutics. In this project we seek to provide a molecular handle to explain some of these fundamental questions, by addressing a potentially under-explored source of immune regulation represented by epigenetic mechanisms. Our paradigm-shifting hypothesis centers on the epigenetic regulation of the immune response implicated in the pathogenesis of AD, and on a newly identified class of long noncoding RNAs (lncRNAs), the immune gene priming lncRNAs (IPLs), that exploit pre-formed chromatin topology at specific cytokine nuclear compartments to facilitate their epigenetic priming and activation. We propose to test and expand our hypothesis using technological advances that only now make this possible. I outline here a plan to pursue this opportunity with 3 specific aims. In Aim 1 (the R61 phase), we will establish the upstream molecular events that coordinate the epigenetic state of inflammatory genes. We hypothesize that IPLs are critical mediators that directly interact with the 3-dimensional chromatin architecture to prime chemokine promoters and enhance the pro- inflammatory responses in innate immune cell activation. We will characterize the mechanisms through which IPLs regulate IL-4 and IL-13, keystone interleukins critical to the induction and perpetuation of the Type 2 response in AD. In Aim 2, we will identity novel protein components that directly interact with the IPLs to modulate the chromatin landscape. We will also perform chromosome conformation capture across the IL-4/IL-13 locus to establish whether changing levels of IPL-IL4/13 alters chromosomal looping across the locus. Lastly, in Aim 3 (R33 phase) we will test inhibitors that specifically target IPL-4/13 in established murine AD-like models that recapitulate human AD. The IPL inhibitors will also be evaluated for their effects on the sensory responses known to influence AD behavior, to modulate direct neuronal priming by Type 2 cytokines shown to be a key step in the pathogenesis of AD. Taken together, our data will directly establish how IPLs and 3-dimensional chromatin architecture act in a cooperative manner to reduce gene-intrinsic noise, and allow robust activation of innate immune genes. A more precise fine-tuning of chemokine transcription through direct manipulations of IPL activities represents a highly valuable therapeutic strategy to achieve tailored immunomodulation in AD.