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

特应性皮炎（AD）是世界范围内最常见的慢性炎症性皮肤病， 由终末角质形成细胞分化缺陷和强2型免疫缺陷驱动。 应答什么控制AD疾病活动？为什么它对治疗的反应不同于 病人对病人？这些都是知之甚少的问题，并提出了一个巨大的未满足的需求， 既有效又安全的治疗方法。在这个项目中，我们试图提供一个分子手柄， 通过解决一个潜在的未充分探索的问题， 以表观遗传机制为代表的免疫调节来源。我们的范式转变 这一假说的中心是免疫应答的表观遗传调节， AD的发病机制，以及一类新发现的长链非编码RNA（lncRNA）， 免疫基因引发lncRNA（IPL），其利用特定位置的预形成的染色质拓扑结构， 细胞因子核区室，以促进其表观遗传引发和激活。我们提出 来测试和扩展我们的假设，利用技术进步， 可能我在这里概述了一个计划，以追求这个机会，有3个具体目标。目标1（R61） 阶段），我们将建立上游分子事件，协调表观遗传状态， 炎症基因我们假设IPL是直接与 三维染色质结构，以总理趋化因子启动子和增强前， 先天免疫细胞激活中的炎症反应。我们将描述这些机制 通过这些途径，IPL调节IL-4和IL-13，这两种关键的白细胞介素对诱导和 AD中2型反应的延续。在目标2中，我们将确定新的蛋白质组分 直接与IPL相互作用以调节染色质景观。我们还将表演 在IL-4/IL-13基因座上捕获染色体构象，以确定是否改变 IPL-IL 4/13水平改变了染色体在基因座上的成环。最后，在目标3（R33阶段）中， 我们将在已建立的小鼠AD样模型中测试特异性靶向IPL-4/13的抑制剂， 概括人类AD。还将评价IPL抑制剂对感觉的影响。 已知影响AD行为的反应，以调节2型AD的直接神经元引发 细胞因子被证明是AD发病机制中的关键步骤。综合起来，我们的数据将 直接建立IPL和三维染色质结构如何在一个合作的过程中起作用， 以减少基因内在噪音的方式，并允许先天免疫基因的稳健激活。一 通过直接操作IPL活性更精确地微调趋化因子转录 代表了在AD中实现定制免疫调节的非常有价值的治疗策略。

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.