Reversing the ocular impact of NM and SM through novel therapies

项目摘要/摘要 眼前表面与泪膜一起提供了防止渗透的第一道防线。 有毒物质，这会对视力造成不利影响。一旦这个两个组成部分的系统被打破， 前表面能够产生强烈的反应，这涉及到相对较快的恢复 上皮化，伴随着活跃的炎症发作。前段反应的一个变异体 患者在接触化学战剂硫磺芥末(SM)后会出现不安。这个 急性症状通常在2-6周后完全消失，不会有进一步的炎症。然而，在一个子集中 在患者中，角膜上皮侵蚀，角膜缘缺血，偶尔还有周边角膜变薄和 新生血管可能进展缓慢。关于治疗方式，无论是类固醇还是非类固醇 消炎药是治疗急性期和长时间的公认的治疗方法。然而，从长远来看， 在SM眼损伤中使用局部类固醇激素并不理想。因此，预防眼科疾病的其他治疗方法 SM受害者的恶化是一个主要的未得到满足的需求。我们最近展示了局部应用 人工合成的功能性高密度脂蛋白纳米粒(HDLNPs)固有地促进再上皮化 在角膜损伤后，可作为化学烧伤后的消炎剂。 此外，高密度脂蛋白纳米粒是将miRNAs运送到角膜的有效载体。因此，高密度脂蛋白NPs 将与角膜中一种高度抑制血管的miRNA miR-184复合，以治疗NM的延迟期- 致角膜损伤。我们也有证据表明维生素D3(VitD3)和聚(乳酸-羟基乙酸)免疫 系统修饰纳米粒(PLGA-IMPS)或局部应用高密度脂蛋白纳米粒，给小鼠 暴露于局部氮芥末(NM)，一种SM的类似物，显著减少促炎和 血管生成信号。总的来说，在HDL、VitD3、miR-184和PLGA-IMP上的这些观察结果具有广泛的 翻译潜力作为SM致眼损伤的新治疗方式。为了测试这一想法，我们将 针对NM对小鼠的急性和延迟期损伤，开发和评估这些局部眼科疗法。 我们将评估这些治疗方法的疗效和机制，结合临床， 形态、细胞生物学和免疫学方法。此外，我们还将评估一类新的高密度脂蛋白 NPS，用有机的、透明的脂质结合的核心支架取代无机的Au核心，这将 不会阻碍光的通过。我们还将评估系统地给药维生素D3或PLGA的效用- IMPS治疗NM所致眼损伤的延迟期这些结果将建立最有效的 NM损伤急性期和延迟期的局部和全身治疗方案。最后，我们将应用这个 了解我们选择的治疗方法对兔眼SM损伤的疗效 描述为NM损伤。最终，我们的研究将为SM引起的损伤提供创新的治疗方法 具有抗炎、抗血管生成、促进分解的能力，副作用最小。

PROJECT SUMMARY/ABSTRACT The ocular anterior surface in conjunction with the tear film provide the first line of defense against penetration of noxious agents, which can adversely affect vision. Once this two-component system is breached, the anterior surface is capable of mounting a vigorous response, which involves a relatively rapid re- epithelialization along with a brisk inflammatory episode. A variant on the response of the anterior segment to perturbation occurs to patients following exposure to the chemical warfare agent, sulfur mustard (SM). The acute symptoms usually resolve completely without further inflammation after 2-6 weeks. However, in a subset of patients, corneal epithelial erosions, limbal ischemia, and, occasionally, peripheral corneal thinning and neovascularization may slowly progress. With respect to treatment modalities, either steroidal or non-steroidal anti-inflammatory drugs are the accepted treatment for the acute and prolonged phases. However, long-term use of topical steroids in SM ocular injuries is not ideal. Thus, additional therapies to prevent the ocular deterioration of SM victims is a major unmet need. We have recently demonstrated that topical application of synthetic, functional high-density lipoprotein nanoparticles (HDL NPs) inherently enhance re-epithelialization following corneal wounding and can act as anti-inflammatory agents following a chemical burn to the cornea. Furthermore, HDL NPs are effective vehicles for the delivery of miRNAs to the cornea. Therefore, HDL NPs will be complexed with miR-184, a highly angiostatic miRNA in the cornea, to treat the delayed phase of NM- induced corneal injury. We also have evidence that Vitamin D3 (Vit D3) and poly(lactic-co-glycolicacid) immune modifying nanoparticles (PLGA-IMPs) systemically or HDL NPs topically, administered to mice that were exposed to topical nitrogen mustard (NM), an analogue of SM, significantly reduce pro-inflammatory and angiogenic signaling. Collectively, these observations on HDLs, Vit D3, miR-184 and PLGA-IMPs have vast translational potential as novel treatment modalities for SM-induced injury to the eye. To test this idea, we will develop and assess these topical ocular therapeutics for the acute and delayed phases of NM injury to mice. We will evaluate the efficacy and mechanisms underlying these treatments with a combination of clinical, morphological, cell biological and immunological approaches. Additionally, we will evaluate a new class of HDL NPs that replace the inorganic Au core with an organic, transparent lipid-conjugated core scaffold, which will not inhibit the passage of light. We will also appraise the utility of systemic administration of Vit D3 or PLGA- IMPs to treat the delayed phase of NM-induced ocular injury. These results will establish the most effective topical and systemic regimens to treat the acute and delayed phases of NM injury. Finally, we will apply this knowledge to evaluate the efficacy of our chosen treatments for SM injury in rabbit eyes using the approaches described for NM injury. Ultimately, our studies will yield innovative treatments for SM-induced injury that will have anti-inflammatory, anti-angiogenic, pro-resolving, capabilities with minimal side effects.