Corneal neuropeptides and herpetic stromal keratitis

角膜神经肽与疱疹性基质角膜炎

• 批准号: 8812856
• 负责人: Susmit Suvas
• 金额: $ 37.41万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-03-01 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8812856
• 关键词: Address; Affect; Afferent Neurons; Agonist; Alcohol or Other Drugs use; Antibodies; Apoptosis; Biological Assay; Blindness; C57BL/6 Mouse; Capsaicin; Cells; Chronic; Clinical; Cornea; Corneal Injury; Dendritic Cells; Detection; Development; Dichloromethylene Diphosphonate; Epidermal Growth Factor; Goals; Health; Herpesvirus 1; Herpetic Keratitis; Immune; In Situ Nick-End Labeling; Infection; Inflammation; Inflammatory; Interleukin-12; Keratitis; Knowledge; Learning; Lesion; Liposomes; Measures; Mediating; Migration Assay; Mus; Natural Killer Cells; Nerve; Nerve Fibers; Nerve Growth Factors; Neuropeptides; Pathogenesis; Peptides; Pharmacologic Substance; Property; Reporting; Research; Role; Severities; Staining method; Stains; Substance P; TACR1 gene; Testing; Time; Topical application; Viral; Viral Load result; Virus; Work; Wound Healing; base; cell type; cytokine; cytotoxicity; in vivo; innate immune function; macrophage; migration; monocyte; mouse model; novel; novel strategies; pre-clinical; receptor; research study; response; tissue repair

DESCRIPTION (provided by applicant): Corneal infection with herpes simplex virus-1 (HSV-1) causes herpetic stromal keratitis (HSK), a leading cause of infection-induced corneal blindness worldwide. Despite intensive research and substantial progress in understanding the pathogenesis of HSK, the management of this condition continues to be challenging. One significant hurdle is our poor understanding of the role of neuropeptides, secreted by abundant corneal nerve fibers, in the development of HSK lesions. Lack of such knowledge is an important problem because neuropeptides are well known to regulate both inflammation and tissue repair. The objective of this application is to determine what regulates substance P (SP) neuropeptide levels in the cornea after ocular HSV-1 infection, and how SP interactions with its receptor NK1R control the viral load and affects the development of HSK lesions. We will approach these questions in a mouse model by using SP-/- and NK1R-/- mice. We recently reported that blocking SP-NK1R interactions in the absence of replicating virus (clinical period) significantly reduced the development of HSK lesions. However, lack of SP-NK1R interactions during active viral replication in the cornea (pre-clinical period), enhances the severity of HSK lesions, as noted in SP- /- and NK1R-/- mice. Our preliminary results showed delayed viral clearance and reduced levels of epidermal growth factor (EGF), a corneal wound healing factor, in NK1R-/- mice. On the basis of our preliminary results, we hypothesize that SP, produced in HSV-1 infected corneas by macrophages and corneal nerves, promotes corneal tissue repair and regulates the influx, survival, and function of innate immune cells involved in viral clearance therefore, enhancing SP-mediated effects during active viral replication should reduce the development of severe HSK lesions. Three aims are proposed to address our hypothesis. In aim 1, experiments will be carried out to determine the role of corneal macrophages and sensory neurons in regulating the levels of SP peptide in HSV-1 infected corneas during the pre-clinical and clinical periods of HSK, respectively. In aim 2, experiments will be carried out t determine whether SP-NK1R interactions regulate viral load in the cornea by promoting the migration, survival, and functions of dendritic cells (DCs), inflammatory monocytes (IM) and natural killer (NK) cells in HSV-1 infected cornea. In aim 3, we will determine whether enhancing SP-mediated effects during the active viral replication period through topical application of EGF alone, or in combination with subconjunctival administration of SP, modulates the development of severe HSK lesions. The information generated by this study could aid in the development of novel pharmaceutical strategies to reduce HSV-1 induced chronic inflammation by promoting viral clearance and corneal tissue repair.

描述（由申请人提供）：单纯疱疹病毒 1 (HSV-1) 的角膜感染会导致疱疹性基质角膜炎 (HSK)，这是全世界感染引起的角膜失明的主要原因。尽管在了解 HSK 发病机制方面进行了深入研究并取得了重大进展，但这种疾病的治疗仍然具有挑战性。一个重要的障碍是我们对丰富的角膜神经纤维分泌的神经肽在 HSK 病变发展中的作用了解甚少。缺乏此类知识是一个重要问题，因为众所周知，神经肽可以调节炎症和组织修复。本应用的目的是确定眼 HSV-1 感染后角膜中 P 物质 (SP) 神经肽水平的调节因素，以及 SP 与其受体 NK1R 的相互作用如何控制病毒载量并影响 HSK 病变的发展。我们将使用 SP-/- 和 NK1R-/- 小鼠在小鼠模型中解决这些问题。我们最近报道，在没有复制病毒的情况下（临床期）阻断 SP-NK1R 相互作用可显着减少 HSK 病变的发展。然而，正如 SP-/- 和 NK1R-/- 小鼠中所指出的，在角膜中活跃的病毒复制过程中（临床前阶段）缺乏 SP-NK1R 相互作用，会增加 HSK 损伤的严重程度。我们的初步结果显示，NK1R-/- 小鼠的病毒清除延迟，表皮生长因子 (EGF)（一种角膜伤口愈合因子）水平降低。根据我们的初步结果，我们假设 SP 由巨噬细胞和角膜神经在 HSV-1 感染的角膜中产生，促进角膜组织修复并调节参与病毒清除的先天免疫细胞的流入、存活和功能，从而增强病毒活跃复制过程中 SP 介导的作用应该会减少严重 HSK 病变的发展。提出了三个目标来解决我们的假设。在目标1中，将进行实验以确定HSK临床前和临床期间角膜巨噬细胞和感觉神经元分别在调节HSV-1感染的角膜中SP肽水平的作用。在目标 2 中，将进行实验以确定 SP-NK1R 相互作用是否通过促进树突状细胞 (DC)、炎症单核细胞 (IM) 和自然杀伤 (NK) 细胞的迁移、存活和功能来调节角膜中的病毒载量在 HSV-1 感染的角膜中。在目标 3 中，我们将确定是否通过单独局部应用 EGF 或与结膜下施用 SP 相结合来增强活跃病毒复制期间 SP 介导的作用，从而调节严重 HSK 病变的发展。这项研究产生的信息可以帮助开发新的药物策略，通过促进病毒清除和角膜组织修复来减少 HSV-1 诱导的慢性炎症。