Project Portugal 2030

Summary

MabExpress aims to help fight one of the most challenging pathogens in human health: Mycobacterium abscessus. To this end, we will develop an innovative, ambitious and complete pre-clinical workflow for discovering and developing new drugs against Mab. Increasing worldwide, Mab infections are hard to treat due to intrinsic antibiotic resistance, complex pathogenesis, and lack of adequate models for drug screening. Thus, improving drug screening and its translation is crucial to better patient outcomes and fight antibiotic resistance. Nevertheless, this poses significant challenges. How we tackle them directly shapes the objectives of this project. Thus, we aim to: a) IMPROVE bacterial quantification: Our Mab double-reporter strains are a novel tool that enables more accurate, efficient, non-invasive, and non-terminal quantification of bacterial load (Annex). This significantly differs from traditional, more laborious, time-consuming methods, representing a leap forward in microbial research and drug screening efficiency. b) REFINE in vitro drug screening: Initial screenings include not only planktonic assays but also simple biofilms and host cell infection– giving us an idea of the drug’s efficacy in different bacterial states. c) SHIFT to high-throughput screenings: High-throughput screenings are key to drug discovery by simultaneously testing thousands of compounds with less preparation time than a manual 96-well assay. It also saves cells and compounds by miniaturising to 384-well microplates (or lower). d) PROGRESS to automatisation: High-throughput assays are a valuable tool, but appropriate technology is required to implement them. Our institution continuously invests in cutting-edge technology, including the state-of-the-art robotic cell::explorer platform, enabling increased throughput and efficiency. e) INTEGRATE different protocols into one workflow: MabExpress proposes a pre-clinical workflow that integrates a series of interconnected stages, from in vitro to in vivo settings. This sequential multi-step approach allows us to confidently select the most promising drug candidates for further development, improving the translatability of pre-clinical findings. f) DEVELOP complex non-mammal models: Mab pathogenesis is characterised by complex 3D structures such as biofilms and granulomas. Thus, we will introduce a crucial step between simple in vitro assays and mice testing with complex infection models: biofilms formed on cystic fibrosis mucus and G. mellonella larvae. g) REDUCE mouse testing: Using our double-reporter strains will reduce the number of animals used in our screening and improve their welfare. The most promising compounds that go through the whole pipeline will be tested in a single mouse model of chronic Mab infection. h) FOSTER new leads against Mab: This pipeline will be used to screen libraries we already have access to through our Consultants. We will also continue to establish collaborations within academia and industry to increase the chances of developing drugs suitable for further development in the drug discovery process. MabExpress will improve, refine, shift, progress, integrate, develop, reduce (animals), and foster the field of drug screening against Mab, advancing our knowledge of infectious diseases. Its ambitious objectives tackle the necessary challenges to advance our ability to combat Mab infections.