SPLUNC1 and Neutrophilic Inflammation in Cystic Fibrosis

我们K 01提案的最初前提是气道宿主防御蛋白短腭肺鼻 上皮细胞克隆-1（PLC 1，SPLUNC 1）调节炎症，因此可能导致急性肺损伤。 损伤（ALI）和CF和呼吸道感染中的组织损伤。我们定义了一种新的免疫调节作用， SPLUNC 1在增强LPS诱导的IFNγ/IFNλ相关炎症中的作用，这使我们重新关注过去几年的研究。 在甲型流感病毒（IAV）肺炎模型上获得K 01奖，其中IFN应答至关重要。这导致 我们提交了R 01提案“A型流感感染期间SPLUNC 1控制的ALI机制”。 IAV死亡主要是由于呼吸衰竭，由肺部炎症和ALI引起。因此 更好地理解IAV引起的肺部炎症和ALI的机制是一个关键的未满足的问题， 需要的IAV激活内体Toll样受体（TLR 3/7/8/9）以诱导气道中的干扰素（IFN）， 免疫细胞，驱动炎症并导致ALI。这一过程的机制并不完全 目前还没有具体的治疗方法来预防ALI或加速其消退。 我们最近发现，气道宿主防御蛋白PLG可能会增加肺部炎症， 通过增强IFNλ的表达来治疗ALI。IFNλ通过IFN刺激基因增加免疫细胞募集， 增加肺部炎症和ALI。磷脂酶可通过充当核酸的支架来调节IFN， 免疫信号蛋白，类似于具有TLR结合基序的其他宿主防御肽。Planted有dual 免疫调节特性和抗病毒作用，减少早期上皮炎症，但可能 在临床过程中增强肺部炎症和ALI。 在最近的工作中，我们发现Plunc-/-小鼠的IFNλ诱导功能受损，而IFNλ诱导功能可以保护它们免受LPS的侵害。 诱发肺部炎症和ALI。我们的初步数据显示，Plunc-/-小鼠受到类似的保护， 在IAV感染过程中，PLR可能通过与TLR 3的相互作用调节这种作用。我们将测试 假设磷脂酰肌醇通过与TLR 3相互作用调节肺部炎症， PLUNC控制的炎症减少了IAV感染期间的ALI。我们将定义PLUNC控制的ALI 可以调节以降低IAV的发病率和死亡率的机制。我们会完成的 通过这些目标：目标1。明确PLC 1增加IFN应答和肺功能的机制 IAV感染期间的炎症;目的2.确定抑制血小板减少性紫癜以防止 IAV-ALI并加速其恢复;以及目标3。定义PLUNC调节的免疫应答和单细胞 与ALI相关的转录组谱及其在人类中的消退。了解PLUNC-TLR 3的作用 在ALI中的相互作用将提供限制IAV发病率的治疗靶点。这里提出的机制提供了 这是限制IAV对数百万人影响的发展机会。我们将使用这个 扩展以支持此提案的初步数据生成，因COVID-19大流行而中断。

The original premise of our K01 proposal was that airway host-defense protein Short Palate Lung Nasal epithelium Clone-1 (PLUNC, SPLUNC1) regulates inflammation and therefore could contribute to acute lung injury (ALI) and tissue damage in CF and respiratory infections. We defined a novel immunomodulatory effect of SPLUNC1 in enhancing LPS-induced IFNγ/IFNλ-associated inflammation that led us to refocus the last years of the K01 award on an Influenza A virus (IAV) model of pneumonia, where IFN responses are critical. This led to the submission of our R01 proposal "SPLUNC1-controlled mechanisms of ALI during influenza A infection". IAV mortality is largely due to respiratory failure, precipitated by lung inflammation and ALI. Therefore, a better understanding of the mechanisms that drive lung inflammation and ALI caused by IAV is a critical unmet need. IAV activates endosomal Toll-Like Receptors (TLR3/7/8/9) to induce Interferons (IFN) in airway and immune cells, driving inflammation and causing ALI. The mechanisms underlying this process are not fully understood and there are no specific therapies to prevent ALI or accelerate its resolution. We recently discovered that airway host defense protein PLUNC may increase lung inflammation and ALI by enhancing IFNλ expression. IFNλ increases immune cell recruitment through IFN-stimulated genes, that increase lung inflammation and ALI. PLUNC may modulate IFN by serving as a scaffold for nucleic acids and immune signaling proteins, similar to other host defense peptides with TLR-binding motifs. PLUNC has dual immunomodulatory properties and antiviral effects that decrease early epithelial inflammation but may enhance lung inflammation and ALI later in the clinical course. In recent work we showed that Plunc-/- mice have impaired IFNλ induction that protects them from LPS- induced lung inflammation and ALI. Our preliminary data now show that Plunc-/- mice are similarly protected during IAV infection and that PLUNC may modulate this effect through interactions with TLR3. We will test the hypothesis that PLUNC regulates lung inflammation through interactions with TLR3, and that blocking PLUNC-controlled inflammation decreases ALI during IAV infection. We will define PLUNC-controlled ALI mechanisms that can be modulated to decrease the morbidity and mortality of IAV. We will accomplish this through these aims: Aim 1. Define the mechanisms by which PLUNC increases IFN responses and lung inflammation during IAV infection; Aim 2. Define the therapeutic potential of inhibiting PLUNC to protect against IAV-ALI and accelerate its recovery; and Aim 3. Define PLUNC-regulated immune response and single-cell transcriptome profiles associated with ALI and its resolution in humans. Understanding the role of PLUNC-TLR3 interactions in ALI will provide therapeutic targets to limit IAV morbidity. The mechanisms proposed here offer an opportunity for developments that would limit the impact of IAV on millions of individuals. We will use this extension to support preliminary data generation for this proposal, disrupted by the COVID-19 pandemic.