Molecular Targets of Corneal Antiangiogenesis

角膜抗血管生成的分子靶点

基本信息

批准号：
8114009
负责人：
ROYCE MOHAN
金额：
$ 36.36万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-08-01 至 2012-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8114009
关键词：
Affinity Alkalies Angiogenesis Inhibitors Antigen Presentation Attenuated Binding Binding Proteins Biochemical Biochemical Pathway Biological Factors Blindness Blood Vessels Burn injury Cell Culture Techniques Cell Cycle Cell Cycle Arrest Cells Chemicals Collagen Competence Cornea Corneal Neovascularization Cytostatics Data Differentiation Antigens Dose Drug Design Endothelial Cells Fibroblasts Genetic Programming Healed Homeostasis Inflammation Inflammatory Injury Investigation Lactones Lead Link Mediating Mediator of activation protein Modeling Molecular Molecular Target Mus NF-kappa B Natural regeneration Pathway interactions Pattern Pharmaceutical Preparations Phenotype Process Production Protein Binding Protein Inhibition Proteins Quality Control Reaction Time Reagent Regulation Research Personnel Role Signal Transduction Signaling Protein Stromal Cells Testing Tissues Translational Research Transplantation Ubiquitin Ubiquitination Vascular Endothelial Cell Vimentin Wound Healing analog angiogenesis antiangiogenesis therapy cooking healing in vivo inhibitor/antagonist injured mouse model multicatalytic endopeptidase complex neovascularization new growth novel novel therapeutic intervention prevent programs repaired research study small molecule transcription factor ubiquitin isopeptidase

项目摘要

DESCRIPTION (provided by applicant): The leading cause of corneal blindness world wide is from angiogenesis, which is the pathogenic growth of new blood vessels from an existing vasculature. Normal wound healing in the cornea does not involve angiogenesis. Therefore, identifying the molecular mechanisms that are responsible for preventing self- destructive angiogenic processes have important bearing on corneal homeostasis, regeneration, and transplantation. A novel biochemical feature preventing corneal stromal cells from participating in pathogenic fibrotic repair processes has recently been linked to the evolutionary conserved ubiquitin proteasome pathway (UPP). The UPP regulates protein quality control, inflammatory transcription factor NF-kappa B activation, cell cycle, differentiation and antigen presentation. We have investigated the hypothesis that a DPP-mediated genetic program controls the angiogenic phenotype of the cornea. We show that the UPP is upregulated in vascular endothelial cells when activated by angiogenic stimulators and during injury healing in vascularized corneas in vivo. This pattern of UPP activation is attenuated by the potent small molecule angiogenesis inhibitor, withaferin A. To demonstrate that a protein binding target(s) of withaferin A mediates this drug's anti-angiogenic mechanism, we generated a biotinylated analog of withaferin A. We have successfully exploited this affinity reagent and isolated the withaferin binding target. In this R01 proposal, we plan to exploit angiogenic and nonangiogenic models of wound healing to define the UPP-driven healing mechanism(s) of the cornea and validate newly identified molecular targets for anti-angiogenesis. For these investigations, withaferin A will serve both as a pharmacological agent and a cell permeable probe of its binding-protein target's function. Specifically, we will (1) characterize the drug's inhibitory mode of action on targeting the angiogenic activation of the UPP, (2) investigate key components of the UPP as mediators of the drugs activity, and (3) investigate withaferin-protein target-deficient mouse models to validate the requirement for drug-protein interaction in the corneal anti-angiogenic mechanism of WFA. We believe these investigations focused on the UPP and the target of withaferin A will enable us to reach our long-term objectives of discovering new therapeutic approaches to control angiogenesis and promote its homeostasis.

描述（由申请人提供）：全球角膜失明的主要原因是血管生成，这是现有脉管系统的新血管的致病性生长。角膜中的正常伤口愈合不涉及血管生成。因此，确定负责预防自我破坏性血管生成过程的分子机制对角膜稳态，再生和移植具有重要影响。一种新型的生化特征，可防止角膜基质细胞参与致病性纤维化修复过程，最近已与进化保守的泛素蛋白酶体途径（UPP）联系起来。 UPP调节蛋白质质量控制，炎症转录因子NF-kappa B激活，细胞周期，分化和抗原表现。我们研究了以下假设：DPP介导的遗传程序控制角膜的血管生成表型。我们表明，当血管生成刺激剂激活血管内皮细胞和体内血管骨骼损伤时，UPP在血管内皮细胞中上调。这种UPP激活的模式被有效的小分子血管生成抑制剂（用脂蛋白A）衰减。证明了用脂蛋白A的蛋白结合靶标介导了该药物的抗血管生成机制，我们产生了与AFERIN A的生物素化合物A。在此R01提案中，我们计划利用伤口愈合的血管生成和非血管生成模型，以定义角膜的UPP驱动的愈合机制，并验证新鉴定的抗血管生成分子靶标。在这些研究中，伴随着大蛋白A将作为药理学剂和可渗透性探针的结合蛋白靶标的功能。具体而言，我们将（1）将药物在靶向UPP的血管生成激活方面的抑制作用方式，（2）研究UPP的关键组成部分是药物活性的介体，以及（3）使用AFERIN蛋白靶标性缺陷型小鼠进行研究，以验证在核蛋白相互作用中对核蛋白相互作用的核蛋白在核蛋白质抗血管抗生素机制中的需求。我们认为，这些调查的重点是UPP及其witaferin A的目标，这将使我们能够达到发现新的治疗方法来控制血管生成并促进其稳态的目标。