Abstract

Lung diseases are a leading cause of mortality worldwide. Dysregulation of immunomodulatory molecules plays a key role in many pulmonary diseases, including lung cancer, idiopathic pulmonary fibrosis (IPF) and infections. Systemic treatment with immunomodulatory molecules however, can have several drawbacks and include toxic side effects in other organs, the need for continuous delivery and a high cost of production. Similarly, treating immunomodulatory repercussions such as telomere shortening or abnormal miRNA expression in target cells is not easy due to the lack of a technology that efficiently and specifically delivers RNA . Furthermore, viral transformation can result in toxicity and is associated with high costs. To circumvent these problems, we aim to engineer the genome-reduced lung bacterium Mycoplasma pneumoniae as a vector to locally express immunomodulatory proteins in the lung. MPN does not have a cell wall, it directly releases secreted biomolecules into the medium, it does not recombine, it has a unique genetic code that prevents the transfer of genes to other bacteria and we have a non-pathogenic engineered version of it. After getting one of the most comprehensive –omics analysis and data integration done on a bacteria we have engineered a MPN chassis for human therapy. We have shown that our engineered chassis is non-pathogenic in mouse mammary glands, intradermal and in the lung and that it can be used to dissolve biofilms made by S. aureus in intradermal catheter as well as to reduce mouse mortality in lung infections caused by Pseudomonas aerugynosa.