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.

项目摘要 研究思路/原理：角膜穿刺伤或撕裂伤引起的角膜瘢痕（或纤维化）是 导致失明的主要病因。目前，没有一种潜在的角膜纤维化治疗方法是FDA- 但没有一种药物能逆转现有的角膜瘢痕。因此，找到一种治疗方法， 减少或控制瘢痕形成的能力，但这不是侵入性的或具有最小的副作用，仍然是一种 未满足的需求来自体内和体外模型的研究表明，转化生长因子- TGF-β 1（TGF-β 1）是切口损伤后角膜瘢痕形成的关键因素之一。然而，TGF- β 1不是纤维化疾病的理想治疗靶点，这是由于其在正常生理学过程中的重要性。 过程（例如，扩散和迁移）。 目的/假设：在我们的初步体外研究中，我们发现TGF-β抑制剂， β/p38 MAPK（p38）信号通路SB 202190几乎完全阻断人角膜瘢痕形成。 这一发现使我们研究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，肌成纤维细胞是一种涉及瘢痕形成的基质细胞表型。我们 发现p38抑制剂加速了这些肌成纤维细胞向其正常表型的转化。 我们假设p38抑制剂SB 202190可以用作预防和治疗糖尿病的局部治疗剂。 逆转角膜疤痕 具体目标：目标1：SB 202190的安全性评价。目的2：确定 SB 202190预防角膜瘢痕形成。目的3：确定是否可以通过以下方法逆转已形成的角膜瘢痕： SB 202190治疗。 研究设计：在该模型中，将在右角膜中心形成3 mm穿孔切口损伤， Sprague-Dawley大鼠。对于AIM 1，将在伤口形成后立即进行治疗并重新评价 在24小时内，以确保SB 202190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，SB 202190或对照PBS将局部应用1-15天， 如下：滴眼液（局部）每日3次或每3天结膜下注射一次。 对于AIM 3，将允许受伤的角膜愈合14天。将大鼠随机分组并给药 在AIM 1中局部给药0-8周。角膜将被检查混浊和与瘢痕形成相关的蛋白质， 0-14在AIM 2中治疗后10天和在AIM 3中治疗后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Western印迹。混浊和瘢痕蛋白的缺乏或至少显著减少将 表明该抑制剂防止了动物模型中的瘢痕形成。