The development of antimicrobial peptides delivery using POlyhedrin Delivery System (PODS) to treat mucosal ulceration and inflammation

多面蛋白传递系统（PODS）是由多面蛋白三聚体组成的三维稳定的立方晶格，可以保护内部内容物免受环境破坏。它们可以作为开发健壮和多功能纳米颗粒的基础，用于生物技术应用，能够将生物活性蛋白质输送到特定的部位和组织。该项目涉及pod的开发，它可以将抗微生物肽输送到口腔和肠道内皮表面，以加速溃疡和炎症相关的愈合过程。有许多慢性炎症性口腔和胃肠道疾病表现为持续性溃疡，如口腔扁平苔藓、复发性口腔炎（RAS）、复发性多形性红斑、白塞氏病、克罗恩病、溃疡性结肠炎。目前，这些疾病的治疗并不理想，主要依赖于类固醇或生物制剂，这些药物有严重的副作用或非常昂贵。PODS提供了另一种治疗选择，易于制造且具有成本效益。该项目最初将使用含有抗炎IL-10或TGFb的pod，并确定蛋白质传递和对上皮细胞和先天免疫细胞的生物学影响。释放速度和对细胞功能的影响将在一系列不同的条件下确定。抗菌肽的选择将基于文献综述、可获得的多组学数据、与粘膜表面和口腔和胃肠道溃疡性疾病的既往关联。候选肽将被克隆到表达质粒中，纯化并对一组细菌进行活性测试，包括共生和致病染色。研究人员将利用昆虫表达系统将具有抗病原菌活性的抗菌药物插入pod中。抗菌pod将按上述方法进行评估。此外，抗菌pod影响上皮伤口愈合的能力将在不同细菌和免疫原性刺激下使用划痕试验和活细胞成像进行评估。随着项目的成功发展，抗菌pod将被调整和修改，以改变抗菌肽的递送率，提高伤口愈合能力。成功的抗菌pod将推进到溃疡性粘膜疾病的体内模型。该计划将启动一种治疗慢性溃疡性疾病和粘膜炎症的新疗法的早期开发。

POlyhedrin Delivery System (PODS) are composed of polyhedrin trimers organized into a three-dimensional stable cubic lattice that shield the internal content from environmental damage. They can serve as the basis for the development of robust and versatile nanoparticles for biotechnological applications capable of delivering bioactive proteins to specific sites and tissues. The project involved the development of PODS that deliver anti-microbial peptides to oral and gut endothelial surfaces in order to accelerate the healing process associated with ulceration and inflammation. There are a number of chronic inflammatory oral and gastrointestinal conditions that present with persistent ulcers, such as, oral lichen planus, recurrent aphthous stomatitis (RAS), recurrent erythema multiforme, Behcet's, Crohn's disease, ulcerative colitis. Currently the treatments for these diseases are suboptimal and predominantly rely on steroids or biologics, which have severe side effects or are very expensive. PODS offer an alternative treatment option, are easy to manufacture and cost effective. The project will initially use PODS that contain anti-inflammatory IL-10 or TGFb and determine protein deliver and biological impact on epithelial and innate immune cells. The rate of release and the influence on cellular functions will be determined under a range of different conditions. Anti-microbial peptides will be selected on the basis of a literature review, available mulitomic data, previous association with the mucosal surfaces and ulcerative diseases of the oral and gastrointestinal tract. Candidate peptides will be cloned into expression plasmids, purified and activity tested against a panel of bacteria, containing both symbiotic and pathogenic stains. Antimicrobials that demonstrate activity against pathogenic bacterial strains will be taken forward and inserted in PODS using an insect expression system. Anti-microbial PODS will be assessed as above. In addition, the ability of the antimicrobial PODS to influence epithelial wound healing will be assessed using scratch assays and live cell imaging in the presence of different bacterial and immunogenic stimuli. As the project develops successful antimicrobial PODS will be adapted and modified to alter the delivery rate of the antimicrobial peptide and improve the wound healing capability. Successful antimicrobial PODS will be advanced in to in vivo models of ulcerative mucosal disease. The Project will initiate the early stages in the development of a new therapy in the treatment of chronic ulcerative diseases and mucosal inflammation.