VRC Inhibiting p38 to Prevent and Restore Corneal Scarring

VRC 抑制 p38 以预防和恢复角膜疤痕

• 批准号: 10833743
• 负责人: Joseph B. Ciolino
• 金额: $ 25.08万
• 依托单位: SCHEPENS EYE RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10833743
• 关键词: Acceleration; Adverse effects; Aftercare; Animal Model; Back; Blindness; Cicatrix; Cornea; Corneal Injury; Disease; Ensure; Etiology; Evaluation; Eyedrops; FDA approved; Fibroblasts; Fibrosis; Hematoxylin and Eosin Staining Method; Hour; Human; Impaired wound healing; In Vitro; Indirect Immunofluorescence; Injections; Injury; Laceration; Mitogens; Modeling; Myofibroblast; Perforation; Phenotype; Physiological Processes; Proliferating; Proteins; Puncture procedure; Rattus; Research; Research Design; Research Project Summaries; Safety; Signal Pathway; Sprague-Dawley Rats; Stains; Stromal Cells; Surgical incisions; TGFB1 gene; Techniques; Therapeutic; Therapeutic Effect; Toxic effect; Transforming Growth Factor beta; Western Blotting; Wound models; corneal epithelial wound healing; corneal scar; cytotoxicity; healing; in vitro Model; in vivo; inhibitor; kinase inhibitor; migration; p38 Mitogen Activated Protein Kinase; prevent; repaired; therapeutic target; wound

Project Summary Research Idea/Rationale: Corneal scarring (or fibrosis) from corneal puncture injury or laceration is one of the leading etiologies for blindness. Currently, none of the potential treatments for corneal fibrosis have been FDA- approved and none have been shown to reverse an existing corneal scar. Therefore, finding a therapeutic with the ability to reduce or control scarring, but that is not invasive or has minimal adverse effects, remains an unmet need. Studies from both in vivo and in vitro models have demonstrated that transforming growth factor- beta1 (TGF-beta1) is one of the key factors that drive corneal scarring after incision injury. However, TGF- beta1 is not an ideal therapeutic target for fibrotic diseases due to its importance in normal physiological processes (e.g., proliferation and migration). Objective/Hypothesis: During our preliminary in vitro studies, we found that an inhibitor for the TGF- beta/p38MAPK (p38)-signaling pathway, SB202190, almost completely blocked human corneal scar formation. This discovery led us to examine if p38 inhibitor could prevent the transformation of human corneal fibroblasts (normal active stromal cells) into myofibroblasts, a stromal cell phenotype that is involved in scarring. We found that p38 inhibitor accelerated the conversion of these myofibroblasts back to their normal phenotype. We hypothesize that the p38 inhibitor, SB202190, can be used as a local therapeutic for preventing and reversing corneal scarring. Specific AIMS: Aim 1:Safety evaluation of SB202190. AIM 2: Determine the therapeutic effect of the SB202190 to prevent corneal scar formation. AIM 3:Determine if an established corneal scar can be reversed by treatment with SB202190. Study Design: In this model, a 3mm perforating incision injury will be made in the center of the right cornea of Sprague-Dawley rats. For AIM 1, treatment will be applied immediately after wound creation and reevaluated at 24 hours to ensure SB202190 does not cause wound healing delays. Cytotoxicity will also be evaluated in human corneal fibroblasts. For AIM 2, SB202190 or control PBS will be applied locally for 1-15 days as follows: eye drops (topical) will be applied 3 times daily or subconjunctival injection once every three days. For AIM 3, the injured corneas will be allowed to heal for 14 days. Rats will be randomly grouped and treated locally as in AIM 1 for 0-8 weeks. Corneas will be examined for haze and proteins associated with scarring at 0-14 days post-treatment in AIM 2 and 0-8 weeks post-treatment in AIM 3 using the following techniques: 1) Slit lamp and OCT examination; 2) TEM and standard Hematoxylin and Eosin stain 3) Indirect- immunofluorescence and Western blot. Absence, or at least significant decrease, of haze and scar proteins will indicate that the inhibitor prevented scarring in the animal model.

项目概要 研究理念/理由：角膜刺伤或裂伤导致的角膜疤痕（或纤维化）是导致角膜损伤的原因之一。 失明的主要病因。目前，尚未有任何潜在的角膜纤维化治疗方法获得 FDA 批准。 已获得批准，但没有任何一种药物被证明可以逆转现有的角膜疤痕。因此，寻找一种治疗方法 减少或控制疤痕的能力，但不是侵入性的或具有最小的副作用，仍然是一个 未满足的需求。体内和体外模型的研究表明，转化生长因子- beta1 (TGF-beta1) 是切口损伤后导致角膜疤痕的关键因素之一。然而，TGF- 由于 beta1 在正常生理中的重要性，它并不是纤维化疾病的理想治疗靶点 过程（例如增殖和迁移）。 目的/假设：在我们的初步体外研究中，我们发现 TGF-β 抑制剂 beta/p38MAPK (p38) 信号通路 SB202190 几乎完全阻断人角膜疤痕形成。 这一发现促使我们研究 p38 抑制剂是否可以阻止人角膜成纤维细胞的转化 （正常活性基质细胞）转化为肌成纤维细胞，这是一种参与疤痕形成的基质细胞表型。我们 发现 p38 抑制剂加速这些肌成纤维细胞恢复正常表型。 我们假设 p38 抑制剂 SB202190 可用作局部治疗剂来预防和治疗 逆转角膜疤痕。 具体目标： 目标1：SB202190的安全性评价。目标 2：确定治疗效果 SB202190 防止角膜疤痕形成。目标 3：确定是否可以通过以下方法逆转已形成的角膜疤痕： 用SB202190治疗。 研究设计：在该模型中，将在患者的右角膜中央制作一个 3mm 的穿孔切口损伤。 斯普拉格-道利大鼠。对于 AIM 1，将在伤口产生后立即进行治疗并重新评估 24 小时确保 SB202190 不会导致伤口愈合延迟。细胞毒性也将在 人角膜成纤维细胞。对于 AIM 2，SB202190 或对照 PBS 将在局部应用 1-15 天，如下所示： 具体如下：滴眼液（外用）每日3次或每三天一次结膜下注射。 对于 AIM 3，受伤的角膜将需要 14 天的时间来愈合。大鼠将被随机分组​​并接受治疗 局部如 AIM 1 所示，持续 0-8 周。将在以下时间检查角膜是否有混浊和与疤痕相关的蛋白质 AIM 2 治疗后 0-14 天和 AIM 3 治疗后 0-8 周，使用以下技术：1) 裂隙灯和OCT检查； 2) TEM 和标准苏木精和伊红染色 3) 间接- 免疫荧光和蛋白质印迹。雾霾和疤痕蛋白的缺失或至少显着减少将 表明该抑制剂可以防止动物模型中出现疤痕。