Live biotherapeutics for the treatment of nontuberculous mycobacteria lung infection associated with cystic fibrosis

Cystic fibrosisCFis a genetic diseasecausing thick mucus to buildup in the airways that is associated with chronic inflammation and persistent lung infectionultimately leading to respiratory failureNontuberculosis mycobacterialNTMinfectionssuch as Mycobacterium abscessusMabare among the most severe chronic infections associated with CFProlonged treatment with potent antibiotics is often met with low compliance due to the severe side effectscomplicated by frequent recurrence and drug resistant strainsSciBac has developed a novel live biotherapeutic using a proprietary microbial improvement platform called Directed Recombination by In Vitro EvolutionDRIVEthat transfers therapeutic traits to commensal microbial strainsThis proposal focuses on the use of DRIVE technology to combine mucolytic and antibacterial traits to optimize a lactobacillus hybrid to demonstrate the safe and effective use in treating chronic Mab lung infection in a relevant mouse modelThe hypothesis is that the DRIVE platform will generate hybrid strains that produce bacteriocins and metabolize mucin resulting in a stand alone live biotherapeutic to treat chronic NTM lung infectionTo this end we aim to firstoptimize the efficacy of a Lactobacillus delbrueckii hybrid using DRIVE to transform NTM antibacterial activity from Bacillus subtilisThe ability of probiotic strains to survive in a competition rich environment is essential for the sustained delivery of therapeutic metabolitesThe optimize Ldelbrueckii hybrid will be characterized and new activities validated via time kill analysis and qualitative mucin degradation assays in vitroThensafety and stability of optimized Ldelbrueckii hybrids will be validated in a cytotoxicity assay with human lung epithelial cells and cytokine production using primary human bronchial tracheal epithelial cellsnormal and CFevaluated via ELISAThe genetic transfer will be fully characterized through comparative whole genome sequencing of donorhostand hybrid strains and repeated passagePreclinical safety in heathy mice and efficacy studies in a statistically relevant agar bead mouse model of chronic Mab infection will be used to demonstrate proof of conceptFollowing treatmentboth infected and uninfected mice lungs and spleens will be assessed for viable colony forming unitsNTM and or hybridand cytokine analysistotal cell countcell differential analysis of the bronchoalveolar lavage fluid will also be completedThe results will determine initial safety and efficacy in animal models and provide support for future preclinical testingThe goal of this Phase I effort is to demonstrate proof of concept that live biotherapeutics are safe and effective in treating Mab lung infection Cystic fibrosis causes severe chronic lung infections resulting in prolonged treatment with potent with severe side effectscomplicated by frequent recurrence and drug resistant strainsThis project will demonstrate proof of concept of a first in class inhaled live biotherapeutic treatment for chronic lung infections associated with cystic fibrosisspecifically nontuberculous mycobacteriaThis innovative strategy will bring new options that influence better antibiotic stewardshipstop the spread of the diseaseand save lives