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

多角体递送系统(PODS)是由组织成三维稳定立方晶格的多面体三聚体组成的，保护内部内容物免受环境破坏。它们可以作为开发用于生物技术应用的强大和多功能纳米颗粒的基础，能够将生物活性蛋白输送到特定的部位和组织。该项目涉及开发豆荚，将抗菌肽输送到口腔和肠道内皮细胞表面，以加速与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，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，这些疾病的治疗方法并不理想，主要依靠类固醇或生物制剂，它们有严重的副作用或非常昂贵。豆荚提供了另一种治疗选择，易于制造，成本效益高。该项目最初将使用含有抗炎IL-10或TGFb的豆荚，并确定蛋白质传递和对上皮细胞和先天免疫细胞的生物影响。释放速率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、先前与口腔和胃肠道的粘膜表面和溃疡性疾病的相关性。候选多肽将被克隆到表达载体中，纯化并对一组包含共生和致病菌株的细菌进行活性测试。表现出对致病细菌菌株的活性的抗菌剂将被向前推进，并使用昆虫表达系统插入豆荚中。抗微生物豆荚将如上所述进行评估。此外，在不同的细菌和免疫原性刺激下，将使用划痕试验和活细胞成像来评估抗菌豆荚影响上皮伤口愈合的能力。随着该项目的发展，将对成功的抗菌豆荚进行改造和改造，以改变抗菌肽的输送速度，提高伤口愈合能力。成功的抗菌豆荚将被推进到溃疡性黏膜疾病的体内模型中。该项目将启动开发一种治疗慢性溃疡性疾病和粘膜炎症的新疗法的早期阶段。

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.