The role of PPAR{gamma} ligands in corneal wound healing and optics

PPAR{γ}配体在角膜伤口愈合和光学中的作用

基本信息

批准号：
8500289
负责人：
Krystel R Huxlin
金额：
$ 36.69万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-08-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8500289
关键词：
Ablation Acetates Acetylation Adult Adverse effects Animals Antibodies Autopsy Bilateral Biological Assay Blindness Cell Death Cell Differentiation process Cell Proliferation Chemicals Cicatrix Clinical Treatment Clinical effectiveness Collagen Cornea Corneal Injury Corneal dystrophy Cytokine Signaling Diabetes Mellitus Dose EP300 gene Effectiveness Elective Surgical Procedures Environment Extracellular Matrix Eye Injuries Felis catus Fibroblasts Fibronectins Fibrosis Genetic Goals Healed Human Image Immunohistochemistry In Situ In Vitro Indium Infection Inflammation Keratoplasty Lasers Ligands Measures Mediating Messenger RNA Mitomycins Modeling Molecular Mus Myofibroblast Nuclear Receptors Operative Surgical Procedures Ophthalmologic Surgical Procedures Optical Coherence Tomography Optics Organ PPAR gamma Pathway interactions Phosphorylation Production Reaction Regulation Relative (related person)Reporting Role Signal Transduction Speed Steroids Structure Testing Time Topical application Trauma Visual Vitamin A Deficiency Western Blotting Work World Health Organization Wound Healing attenuation connective tissue growth factor corneal scar cytokine design effective therapy healing in vivo inhibitor/antagonist insight insulin sensitizing drugs meetings migration novel prednisolone prevent public health relevance research study transcription factor

项目摘要

DESCRIPTION (provided by applicant): Corneal scarring is a major cause of decreased visual quality and vision loss worldwide. Scarring follows disruption to normal corneal structure and function, whether from infection, laser refractive surgery, corneal transplantation, ocular trauma (chemical or physical) or corneal dystrophies. There is no suitable means of controlling corneal scaring despite more than 25 years trying to characterize cytokine signaling during wound healing. Our long-term objective is to understand mechanisms of corneal wound healing and our goal is to design effective therapies to treat or prevent corneal scarring. Our overall hypothesis is that peroxisome proliferator activated receptor gamma (PPAR3) ligands prevent corneal fibrosis and can be targeted as a therapy for this condition, with significantly greater efficacy than current clinical treatments or than blocking the activity of single cytokines. Aim 1: Test the hypothesis that PPAR3 ligands inhibit key pro-fibrotic activities of cultured corneal keratocytes. We use immunohistochemistry, proliferation assays, wounding assays, western blots, slot blots and Q-PCR to assess the relative effectiveness of PPAR3 ligands at modulating proliferation, migration, expression of CTGF, 1SMA, Thy-1, collagen I, collagen III, fibronectin and their mRNAs in cultured keratocytes stimulated by optimal doses of TGF2. Aim 2: Test the hypothesis that PPAR3 ligands inhibit key pro-fibrotic activities in cultured corneal keratocytes through both PPAR3-dependent and -independent pathways. We will use both pharmacological and genetic approaches to test our prediction that PPAR3 ligands act both by activating PPAR3 and by inhibiting TGF2-regulated pathways. Knowing the relative strengths of these mechanisms in corneal keratocytes is critical if we are to develop PPAR3 ligands as optimally-targeted therapies for corneal fibrosis. Aim 3: Test the hypothesis that PPAR3 ligands are more efficient inhibitors of fibrosis in PRK-induced corneal wound healing than anti-TGF2 antibodies, steroids or Mitomycin C. We will perform binocular PRK in cats followed by the topical administration of select PPAR3 ligands, anti-TGF2 antibodies, steroids or Mitomycin C. We will use immunohistochemistry to contrast key cellular aspects of the wound healing reaction. We predict that PPAR3 ligands will be associated with lower cell death, faster wound healing, less haze and lower induction of higher-order optical aberrations than use of anti-TGF2 antibodies, steroids or Mitomycin C post-operatively.

描述（由申请人提供）：角膜疤痕是全球视觉质量和视力丧失降低的主要原因。无论是从感染，激光屈光手术，角膜移植，眼部外伤（化学或物理）还是角膜营养不良的情况下，疤痕都会破坏正常的角膜结构和功能。尽管超过25年试图表征伤口愈合过程中的细胞因子信号传导，但没有合适的方法来控制角膜恐怖。我们的长期目标是了解角膜伤口愈合的机制，我们的目标是设计有效的疗法来治疗或预防角膜疤痕。我们的总体假设是，过氧化物酶体增殖物激活的受体伽马（PPAR3）配体可以防止角膜纤维化，并且可以作为这种疾病的治疗方法，其疗效明显高于当前临床治疗或阻止单细胞因子的活性。目标1：检验PPAR3配体抑制培养的角膜角膜细胞的关键促纤维化活性的假设。我们使用免疫组织化学，增殖测定，伤口测定，蛋白质印迹，插槽和Q-PCR来评估PPAR3配体在调节增殖，迁移，CTGF，1SMA，1SMA，THY-1，THY-1，THY-1，胶原I，胶原I，胶原III，Fibronectin及其MRNASCRATIEN CRICTIS cORTISER的刺激量的相对有效性。 AIM 2：检验以下假设：PPAR3配体通过PPAR3依赖性和非依赖性途径抑制培养的角膜角膜细胞中的关键促纤维化活性。我们将同时使用药理学和遗传方法来测试我们的预测，即PPAR3配体通过激活PPAR3和抑制TGF2调节的途径来起作用。如果要开发出作为角膜纤维化的最佳靶向疗法，知道角膜角膜细胞中这些机制的相对优势至关重要。目标3：检验以下假设：PPAR3配体比抗TGF2抗体，类固醇或丝裂霉素更有效地抑制了PRK诱导的角膜伤口愈合。免疫组织化学与伤口愈合反应的关键细胞方面对比。我们预测，与使用抗TGF2抗体，类固醇或术后脱甲霉素C相比，PPAR3配体将与较低的细胞死亡，更快的伤口愈合，更少的雾霾和更低的高阶光学诱导有关。