Targeting anti-viral and anti-inflammatory responses during ocular HSV-1 infection to prevent vision impairment.

项目摘要 角膜的透明度是正常视力所必需的。然而，单纯疱疹病毒的反复发作 单纯疱疹病毒1型(HSV-1)感染角膜会导致间质角膜炎(SK)，这是一种非常痛苦和视力受损的疾病 慢性炎症性疾病SK是一种严重的疾病，也是导致儿童传染性失明的主要原因 我们。病毒在角膜上皮细胞中的复制和相关的炎症在SK中起着中心作用 进步。目前的SK疗法，如皮质类固醇和抗病毒药物，是非特异性的，部分有效的， 并引起严重的眼部和全身副作用。因此，发展小说有一种未得到满足的需求 免疫疗法，以解决与当前SK疗法相关的缺陷。选择性地诱导一种 强大的抗病毒状态与最低限度的炎症免疫反应的激活体现了一种强有力的手段 治疗复发性慢性SK病理患者。在本申请中，我们提出了一种这样的方法 靶向免疫调节细胞因子IL-27诱导内源性抗病毒和抗炎 角膜HSV-1感染后抑制SK进展的反应。巨噬细胞(MϕS)发挥核心作用 在HSV-1清除中，通过吞噬感染的上皮细胞和凋亡的中性粒细胞。我们的预赛 数据表明，单纯疱疹病毒1型刺激MϕS产生IL-27。此外，我们还表明IL-27在限制 单纯疱疹病毒1型在角膜中的复制，干扰素-β的产生，适应性免疫反应的最佳诱导，以及 抑制SK病理。此外，我们还发现，单纯疱疹病毒1型通过促进M-ϕ代谢重编程 显著增加免疫反应基因1(IRG1)的表达。IRG1是一种能转化柠檬酸的酶 在三羧酸(TCA)循环中转化为衣康酸。衣康酸是一种线粒体代谢物，由 激活MϕS调节炎症和抗病毒反应。我们的数据显示IL-27呈负性 调节单纯疱疹病毒1型感染后M-ϕS细胞中irg1的表达。因此，我们的中心假设是IL-27起双重作用 通过调节HSV-1诱导的升高在SK进展中的抗病毒和抗炎作用 MϕS体内irg1/衣康酸代谢促进I型iFN介导的保护性反应在这项提案中，我们 将探讨IL-27介导的先天和获得性免疫调节的分子机制 SK进展过程中的免疫反应(Aim1)，并阐明HSV-1诱导的IL-27和 M-ϕS的irg1/衣康酸代谢及其在SK进展中的作用(目标2)。圆满完成拟建 研究将确定单纯疱疹病毒1型诱导Mϕ代谢重编程的新分子机制 Mϕ特异性IL-27/IRG-1/衣康酸在诱导抗病毒抗炎性反应中的作用 在SK期间。这些研究将为开发新的、安全的和有效的靶向治疗提供平台 IL-27和Mϕ代谢抑制单纯疱疹病毒1型复发和SK进展。

Project Summary The corneal transparency is essential for normal vision. However, the recurrent episodes of herpes simplex virus-1 (HSV-1) infection of the cornea causes stromal keratitis (SK), a very painful and vision impairing chronic inflammatory disease. SK is a severe condition and the leading cause of infectious blindness in the US. The virus replication in the corneal epithelium and associated inflammation play a central role in SK progression. Current SK therapies such as corticosteroids and anti-virals are non-specific, partially effective, and cause severe ocular and systemic side effects. Therefore, there is an unmet need to develop novel immunotherapies to address deficiencies associated with current SK therapies. The selective induction of a potent anti-viral state with minimal activation of inflammatory immune responses embodies a powerful means to treat patients with recurrent chronic SK pathology. In this application, we propose one such approach targeting IL-27, an immunoregulatory cytokine, to induce endogenous anti-viral and anti-inflammatory responses after corneal HSV-1 infection to suppress SK progression. Macrophages (Mϕs) play a central role in HSV-1 clearance through phagocytosis of infected epithelial cells and apoptotic neutrophils. Our preliminary data indicate that HSV-1 stimulates IL-27 production by Mϕs. Further, we show that IL-27 is critical for limiting HSV-1 replication in the cornea, IFN-β production, optimum induction of adaptive immune responses, and suppression of SK pathology. Moreover, we show that HSV-1 promotes Mϕ metabolic reprogramming with significantly increased immune-responsive gene 1 (Irg1) expression. Irg1 is an enzyme that converts citrate to itaconate during the tricarboxylic acid (TCA) cycle. Itaconate is a mitochondrial metabolite produced by activated Mϕs to regulate inflammatory and anti-viral responses. Our data show that IL-27 negatively regulates Irg1 expression in Mϕs after HSV-1 infection. Thus, our central hypothesis is that IL-27 plays a dual anti-viral and anti-inflammatory role in SK progression through modulation of HSV-1-induced increased Irg1/itaconate metabolism in Mϕs to promote type I IFNs-mediated protective responses. In this proposal, we will investigate the molecular mechanisms for IL-27-mediated immunoregulation of innate and adaptive immune responses during SK progression (Aim1) and elucidate the interplay of HSV-1-induced IL-27 and Irg1/itaconate metabolism in Mϕs and role in SK progression (Aim 2). The successful completion of proposed studies will identify novel molecular mechanisms for HSV-1-induced Mϕ metabolic reprogramming and the contribution of Mϕ-specific IL-27/Irg-1/itaconate in the induction of anti-viral versus inflammatory responses during SK. These studies will open the platform for developing novel, safe, and effective therapies targeting IL-27 and Mϕ metabolism to suppress recurrent HSV-1 infection and SK progression.