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

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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.