Therapy for ocular mustard gas exposure using engineered FGF derivatives

使用工程 FGF 衍生物治疗眼部芥子气暴露

• 批准号: 9001388
• 负责人: David D Eveleth
• 金额: $ 33.43万
• 依托单位: E AND B TECHNOLOGIES, LLC
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-30 至 2017-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9001388
• 关键词: Acute; Alkylation; Beds; Binding; Bioterrorism; Cells; Clinical; Cornea; Corneal Injury; Data; Development; Diffusion; Dose; Effectiveness; Engineering; Epithelial Cell Proliferation; Event; Exposure to; Extracellular Matrix; Eye; Eye Injuries; Eyedrops; Family; Fibroblast Growth Factor; Fibroblast Growth Factor 1; Fluorescein; Foundations; Functional disorder; Gelatinase B; Goals; Government; Growth Factor; Healed; Hematoxylin and Eosin Staining Method; Heparin; In Vitro; Individual; Injury; Inorganic Sulfates; Lesion; Lung; Magnetic Resonance Imaging; Measures; Mechlorethamine; Modeling; Monitor; Mustard; Mustard Gas; Mutation; Natural regeneration; Organ Culture Techniques; Oryctolagus cuniculus; Pain; Peptide Hydrolases; Phase; Population; Positioning Attribute; Process; Property; Proteins; Proteoglycan; Proteolysis; Research; Research Project Grants; Residual state; Resistance; Severities; Site; Skin; Staining method; Stains; Stromal Cells; Sulfhydryl Compounds; Surface; System; TNF-alpha converting enzyme; Testing; Therapeutic; Time; Tissues; Unspecified or Sulfate Ion Sulfates; Vesicants; Vision; Visual; War; Work; Wound Healing; analog; disulfide bond; effective therapy; healing; improved; in vivo; injured; member; migration; neovascularization; ocular surface; programs; public health relevance; research study; response; vapor; weapons; wound

 DESCRIPTION (provided by applicant): Mustard gas (sulfur mustard, SM) and related vesicants have been used as weapons of war and are considered by the US government to be a potential terrorist threat to the civilian population. These agents cause rapid, significant and debilitating injuries to the skin, lungs and eyes. The eye and particularly the cornea is the tissu most sensitive to exposure. SM exposure drives microvesication with Deepithelialization of the corneal surface and results in significant pain and degradation of vision. No effective treatment for vesicant injury is known. Treatments that could reduce the corneal lesions and accelerate healing of the corneal surface would be highly valuable subsequent to an attack. The goal of this research is to produce effective therapeutics for SM corneal injury by using engineered versions of naturally occurring growth factors. These substances, including many members of the fibroblast growth factor (FGF) family, are part of the natural wound healing response and can accelerate the healing of a wide variety of corneal wounds. The efficacy of both endogenously produced and exogenously applied growth factors in the context of SM injury may be limited by the sensitivity of growth factors to inactivation by SM, degradation by proteases induced following injury and/or by diffusion of the growth factors from the surface of the eye and the wound bed. We have produced engineered versions of FGF-1, a key growth factor involved in the healing process, that should overcome many of the limitations of the endogenous protein and facilitate its use as an ocular therapy for mustard gas exposure. This research project will first test engineered FGF-1 (eFGF-1) molecules for efficacy in the rabbit corneal organ culture model to prove the hypothesis that the eFGF-1s can accelerate healing of vesicant injury. Two eFGF-1 molecules that 1) lack any of the free thiols of native FGF-1; 2) have additional stability enhancing mutations including an internal disulfide bond; and 3) do not require heparin co-factors will be evaluated. These derivatives should not be modified by any residual SM, be considerably less susceptible to proteolysis and should be retained in the wound site through interactions with sulfated proteoglycans of the extracellular matrix. The ability of these molecules to accelerate the regeneration and proliferation of the ocular surface and inhibit the expression of wound related markers will be shown. Finally, the ability of the eFGF-1s to accelerate healing in the rabbit eye following SM exposure in vivo will be demonstrated. If this work is successful, these eFGF-1s will be advanced into clinical development for SM injury.

 描述（由申请人提供）：芥子气（硫芥子气，SM）和相关发泡剂已被用作战争武器，被美国政府视为对平民人口的潜在恐怖威胁。皮肤、肺和眼睛受到损伤，尤其是角膜，接触 SM 会导致角膜表面出现微泡，导致严重疼痛。目前尚无针对脓疱性损伤的有效治疗方法，可以减少角膜损伤并加速角膜表面的愈合，这项研究的目标是为 SM 角膜提供有效的治疗方法。这些物质，包括成纤维细胞生长因子（FGF）家族的许多成员，是自然伤口愈合反应的一部分，可以加速多种角膜伤口的愈合。两者的在 SM 损伤的情况下，内源产生和外源应用的生长因子可能受到生长因子对 SM 失活的敏感性、损伤后诱导的蛋白酶降解和/或生长因子从眼睛和眼睛表面扩散的限制。我们已经生产了 FGF-1 的工程版本，这是一种参与愈合过程的关键生长因子，它应该克服内源性蛋白质的许多局限性，并促进其用作芥子气暴露的眼部治疗。将首先测试工程化的 FGF-1 (eFGF-1) 分子在兔角膜器官培养模型中具有功效，以证明 eFGF-1 可以加速疱疹性损伤愈合的假设，这两种 eFGF-1 分子 1) 缺乏任何游离硫醇。天然 FGF-1；2) 具有额外的稳定性增强突变，包括内部二硫键；以及 3) 不需要肝素辅助因子。这些衍生物不应被任何残留物修饰。 SM 对蛋白水解的敏感性相当低，应通过与细胞外基质的硫酸化蛋白聚糖相互作用而保留在伤口部位。这些分子加速眼表再生和增殖并抑制伤口相关标记物表达的能力将。最后，将证明 eFGF-1 在体内 SM 暴露后加速兔眼愈合的能力，如果这项工作成功，这些 eFGF-1 将进入针对 SM 损伤的临床开发。