Corneal wound healing and nerve regeneration

 描述（由申请人提供）：角膜神经对于角膜的健康以及保护眼睛免受外部因素的影响非常重要。这些神经主要是伤害性的，响应于机械刺激、温度变化和/或化学刺激而编码不适和疼痛。疾病、感染和眼科手术都会损害角膜神经，造成疼痛、干眼症、复发性糜烂、混浊甚至失明等长期后果。然而，在临床实践中没有治疗神经功能障碍的有效疗法，将使用我们的光折射角膜切除术（photorefractive keratectomy，PRK）后角膜伤口愈合的猫模型以及体内和体外方法的组合来研究控制成人角膜神经损伤后再生的基本机制。我们的初步数据显示，不同神经层（基质，基底神经丛下，上皮）的神经再支配明显异常，局部抗纤维化药物对神经再生有重要调节作用。这些数据表明肌成纤维细胞对再生神经和/或其相关神经胶质（非髓鞘化雪旺细胞-NMSC）的抑制性影响，导致我们提出以下关于该更新应用的中心假设：响应于大角膜伤口而发生的肌成纤维细胞转化直接抑制神经再生。因此，在早期伤口愈合反应期间阻断肌成纤维细胞分化对于恢复上皮和基质的正常角膜神经支配至关重要。我们将通过以下方式检验这一假设：目的1 -评估肌成纤维细胞分化对角膜神经再生和角膜后NMSC的影响;目的2 -评估阻断肌成纤维细胞分化对角膜神经再生和角膜后NMSC的影响;目的3 -角膜伤口的背景。所提出的实验评估异常角膜神经再支配对角膜光学的长期影响;以及目的4 -表征体外角膜成纤维细胞、肌成纤维细胞和感觉神经元之间的相互作用，并测试肌成纤维细胞通过Sema 3A抑制神经突生长的假设。所提出的神经再生、神经-肌成纤维细胞相互作用及其在伤口愈合过程中的分子机制的系统表征对于开发新的治疗策略以用眼睛治疗角膜伤口以促进最佳神经再生并确保眼表的长期健康至关重要。

 DESCRIPTION (provided by applicant): Corneal nerves are important for the health of the cornea, as well as for protecting the eye from outside elements. These nerves are predominantly nociceptive, coding discomfort and pain in response to mechanical stimulation, temperature change and/or chemical stimulation. Disease, infection and ocular surgery can 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 therapies in clinical practice fo treating nerve dysfunction in the will use our cat model of corneal wound healing after photorefractive keratectomy (PRK) and a combination of in vivo and in vitro approaches to study the basic mechanisms controlling adult, corneal nerve regeneration post-injury. Our preliminary data show clear abnormalities of re-innervation in different nerve layers (stroma, sub-basal plexus, epithelium) and major modulation of nerve regeneration by topical anti-fibrotics. These data suggest an inhibitory influence of myofibroblasts on regenerating nerves and/or their associated glia (non-myelinating Schwann cells - NMSCs), leading us to propose the following central hypothesis for this renewal application: myofibroblast transformation that occurs in response to large corneal wounds directly inhibits nerve regeneration. Thus, blocking myofibroblast differentiation during the early wound healing response is critical for restoring normal corneal innervation to the epithelium and stroma. We will test this hypothesis by: Aim 1 - assessing the impact of myofibroblast differentiation on corneal nerve regeneration and NMSCs after PRK; Aim 2 - assessing the effect of blocking myofibroblast differentiation on corneal nerve regeneration and NMSCs after PRK; Aim 3 - context of corneal wounds. The proposed experiments assessing the long-term impact of abnormal corneal re-innervation on corneal optics; and Aim 4 - characterizing the interactions between corneal fibroblasts, myofibroblasts and sensory neurons in vitro and testing the hypothesis that myofibroblasts inhibit neurite outgrowth via Sema3A. The proposed, systematic characterization of nerve regeneration, nerve-myofibroblast interactions and their molecular mechanisms during wound healing are critical for the development of new therapeutic strategies to treat corneal wounds with an eye to promoting optimal nerve regeneration and ensuring long-term health of the ocular surface.