Corneal wound healing and nerve regeneration

角膜伤口愈合和神经再生

• 批准号: 10386769
• 负责人: Krystel R Huxlin
• 金额: $ 38.44万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10386769
• 关键词: Accidental Injury; Adult; Afferent Neurons; Area; Autopsy; Beds; Bioenergetics; Biological Assay; Blindness; Cell Differentiation process; Chemical Stimulation; Chronic; Clinical; Code; Contact Inhibition; Cornea; Corneal Diseases; Corneal Injury; Cues; Data; Disease; Elements; Ensure; Epithelial; Eye; Felis catus; Fibroblasts; Fibrosis; Functional disorder; Genetic; Genome; Goals; Grant; Health; Histologic; Human; Immunohistochemistry; In Vitro; Infection; Injury; Ligands; Literature; Measures; Mechanical Stimulation; Mediating; Metabolic; Mitochondria; Modeling; Molecular; Myofibroblast; Natural regeneration; Nerve; Nerve Regeneration; Neurites; Neurons; Nociception; Operative Surgical Procedures; Pain; Patients; Peripheral; Pharmacology; Phase; Phenocopy; Photorefractive Keratectomy; Plague; Play; Process; Pyruvate; Reaction; Recurrence; Research; Role; Signal Pathway; Signal Transduction; Specificity; Speed; Stains; TGFB1 gene; Temperature; Testing; Therapeutic; Thiazolidinediones; Time; Time Study; Vision; Work; antifibrotic treatment; behavior in vitro; clinical practice; corneal epithelial wound healing; corneal scar; density; design; effective therapy; experimental study; eye dryness; flexibility; in vivo; in vivo regeneration; inhibitor; nerve supply; neurite growth; novel; novel therapeutic intervention; novel therapeutics; ocular surface; prevent; pyruvate carrier; reinnervation; release factor; response; treatment strategy; troglitazone; wound; wound environment; wound treatment

Abstract Corneal nerves are important for cornea health, and for protecting the eye from outside elements. They are mostly nociceptive, coding discomfort and pain in response to mechanical stimulation, temperature and/or chemical stimulation. Disease, infection and ocular surgery all damage corneal nerves, with long-term consequences in terms of pain, dry eye, recurrent erosions, opacity and even blindness. Yet, there are no effective clinical therapies for nerve dysfunction once fibrosis has occurred. Our prior work allowed us to better understand factors that control myofibroblast differentiation in the large wounds generated in human and cat corneas. This work also led to the observation of a nefarious interaction between wound myofibroblasts and regenerating corneal nerves. Pre-treating the wound area with anti-fibrotic agents can mitigate this problem, but in most cases of accidental injury or infection, patients present after myofibroblast differentiation/fibrosis have occurred. The issue then is no longer preventing fibrosis, but rather overcoming the contact inhibition between nerves and persistent myofibroblasts. We recently found that the PPARg ligand Troglitazone stimulates neurite outgrowth and causes myofibroblasts de-differentiation in vitro and in vivo. Importantly, these effects can be mimicked in vitro by a compound that spares PPARg and targets the mitochondrial pyruvate carrier (MPC). Together with recent literature, our pilot data form a strong premise for critically testing the hypothesis that inhibiting the MPC may facilitate metabolic remodeling in both regenerating nerves and activated myofibroblasts to overcome the blocked reinnervation inherent in later stages of post-injury corneal fibrosis. To test this hypothesis, we will: Aim 1, identify metabolic changes through which inhibiting mitochondrial pyruvate transport promotes neurite outgrowth of peripheral sensory neurons in a simulated wound environment; Aim 2, assess if similar or different mechanisms (as in Aim 1) underlie de-differentiation of corneal myofibroblasts; and Aim 3, contrast the relative efficacy of different mitochondrial modulators for overcoming myofibroblasts’ inhibition on corneal nerve regeneration in vivo. Fundamentally, the proposed research will ascertain if MPC activity is key to overcoming fibrosis and/or its associated inhibition of neurite outgrowth. We will confirm target specificity through genome manipulation in both corneal fibroblasts and peripheral sensory neurons, determine how targeting impacts mitochondrial function, and identify signals downstream of metabolic rewiring that are necessary to mediate these effects. Since the MPC has never been studied in the context of peripheral wounding, our results could unveil new, broadly-relevant aspects of its function. Ultimately, the proposed research aims to unravel novel molecular mechanisms that may play a substantial, hitherto unexplored role in large, established, fibrotic wounds. Defining these mechanisms is necessary if we are to develop new therapies for the treatment of mature corneal wounds with an eye to promoting optimal nerve regeneration and ensuring long-term corneal health and clarity.

抽象的 角膜神经对于角膜健康和保护眼睛免受外部元素的影响很重要。他们是 主要是伤害性的，对机械刺激，温度和/或 化学刺激。疾病，感染和眼部手术均损害角膜神经，长期 在疼痛，干眼，复发性侵蚀，不透明甚至失明方面的后果。但是，没有 发生纤维化后，有效的神经功能障碍的临床疗法。我们先前的工作使我们变得更好 了解控制人和猫产生的大伤口中肌纤维细胞分化的因素 角膜。这项工作还导致观察到伤口肌纤维细胞和 再生角膜神经。用抗纤维化剂预先治疗伤口区域可以减轻此问题， 但是在大多数意外受伤或感染的情况下，肌纤维细胞分化/纤维化后出现的患者 发生了。然后，问题不再防止纤维化，而是克服接触抑制 在神经和持续的肌纤维细胞之间。我们最近发现PPARG配体troglitazone 刺激神经蛋白神经病的生长，并在体外和体内引起肌纤维细胞脱不同。重要的是， 这些效果可以在体外模仿，该化合物可避免PPARG并靶向线粒体 丙酮酸载体（MPC）。与最近的文献一起，我们的飞行员数据构成了批判性测试的有力前提 抑制MPC的假设可能促进神经再生和 激活的肌纤维细胞以克服伤害后角膜后期固有的固有的封闭的重新支配 纤维化。为了检验这一假设，我们将：目标1，确定抑制的代谢变化 线粒体丙酮酸转运可促进模拟中周围感觉神经元的神经落生物的生长 伤口环境； AIM 2，评估脱不同的机制是相似还是不同的机制（与目标1） 角膜肌纤维细胞；和目标3，对比不同线粒体调节剂的相对效率 克服肌纤维细胞对体内角膜神经再生的抑制作用。从根本上讲，提议 研究将确定MPC活性是否是克服纤维化和/或其相关抑制神经蛋白的关键 出生。我们将通过角膜成纤维细胞和 周围感觉神经元，确定靶向如何影响线粒体功能并识别信号 代谢重新布线的下游是介导这些作用所必需的。由于MPC从未有过 在周围获胜的背景下研究了我们的结果，可以揭示其新的，广泛的，与之相关的方面 功能。最终，拟议的研究旨在阐明可能发挥的新型分子机制 在大型，已建立的纤维化伤口中实质性的意外作用。定义这些机制是 如果我们要开发新疗法来治疗成熟的角膜伤口，则有必要 促进最佳的神经再生并确保长期角膜健康和清晰度。