Periodic Reporting for period 1 - INNOTARGETS (Innvative approaches to identification of metabolic Targets for antimicrobials)
Periodo di rendicontazione: 2021-03-01 al 2023-02-28
Antimicrobial resistance (AMR) is a challenge to human health and health-systems worldwide. The project INNOTARGETS aims to train 12 young researchers in highly innovative approaches to identify metabolic drug targets for antimicrobials centered around a collaborative research project with the aims 1) to characterize the in vivo metabolism of Streptococcus suis and Staphylococcus aureus during infection, 2) to improve metabolic modelling approaches for use in antibiotic target discovery and modelling in prediction of toxicity of antibiotics, and 3) to identify metabolic targets that can block expression of selected resistances in Streptococcus suis, Staphylococcus aureus and Escherichia coli or prevent spread of conjugative resistance plasmids.
INNOTARGETS has recruited and trains 12 doctoral students. In the reporting period, genes essential for growth of Streptococcus suise in active porcine serum have been identified using a transposon library and such genes are under further characterization. A Staphuylococcus aureus library is under construtcions and will be used to identify genes which are required for growth during systemic infection and skin infections in mice. In parallel, a study of which proteins are expressed by this bacterium during skin infection has been carried out, and the global proteome is currenly being analysed.
INNOTARGETS has further identified targets, where blocking re-sensitize MRSA Staphylococcus aureus to oxacilling and tetracyline and Eschericiah coli to macrolides and aminoglycoside antibiotics, and work is in progress to identify targets which prevents expression of fosfomycin resistance in Escherichia coli and lincosamid and macrolide resistance in Strepotococcus suis. In all these bacteria, the approach has been to construct transpors libraries to identify genes, where knock-out causes loss of the abilty to grow in the presence of antimicrobials. Further, bioactive molecules have been identified, which block spread of a conjugative plasmids, which encodes reistance to the highly critical antimicrobial for human health, cephalosporin.
There is a high level of redundancy in the metabolism, i.e. more than one enzyme can lead to production of the same metabolite. Such pairs of enzymes may be pututative targets for both antimicrobial and helper drugs, if they are blocked simultaneously. In order to identify such pairs, INNOTARTEGS has constructed mathematical models of the metabolism in Eschericia coli and Staphylococcus aureus. As part of this effort, results from transposon and proteomic studies are build into the modelling in order to generate much more precise modelling. To be able to identify lead molecules which can block pairs of enzymes, one doctoral student is setting up a novel system for screening of libraries of bioactive molecules.
In one of the doctoral projects, a laboratory methods to predict toxicity of drug candidates, and their downstream metabolic by-products has been set up. For validation of the system, 28 commercial drugs with known toxicity profile, belonging to several medical classes, have been assessed in seven cell lines representative of major mamalian body systems and derived from two different animals species and humans.
INNOTARGETS has established a joint training syllabus designed to widen the career prospects of the doctoral students, and to optimize their professional competences. During the training period, technical courses (transposom techniques and proteomes) as well as complementary skills course (data management, scientific writing, IPR rights, entrepreneurship and personal impact) have been offered to the network’s students. INNOTARGETS has communicated preliminary results to the scientific community through participation in international and national meetings and have contributed to public science events centered on the topic of the global antimicrobial crises.
INNOTARGETS has further identified targets, where blocking re-sensitize MRSA Staphylococcus aureus to oxacilling and tetracyline and Eschericiah coli to macrolides and aminoglycoside antibiotics, and work is in progress to identify targets which prevents expression of fosfomycin resistance in Escherichia coli and lincosamid and macrolide resistance in Strepotococcus suis. In all these bacteria, the approach has been to construct transpors libraries to identify genes, where knock-out causes loss of the abilty to grow in the presence of antimicrobials. Further, bioactive molecules have been identified, which block spread of a conjugative plasmids, which encodes reistance to the highly critical antimicrobial for human health, cephalosporin.
There is a high level of redundancy in the metabolism, i.e. more than one enzyme can lead to production of the same metabolite. Such pairs of enzymes may be pututative targets for both antimicrobial and helper drugs, if they are blocked simultaneously. In order to identify such pairs, INNOTARTEGS has constructed mathematical models of the metabolism in Eschericia coli and Staphylococcus aureus. As part of this effort, results from transposon and proteomic studies are build into the modelling in order to generate much more precise modelling. To be able to identify lead molecules which can block pairs of enzymes, one doctoral student is setting up a novel system for screening of libraries of bioactive molecules.
In one of the doctoral projects, a laboratory methods to predict toxicity of drug candidates, and their downstream metabolic by-products has been set up. For validation of the system, 28 commercial drugs with known toxicity profile, belonging to several medical classes, have been assessed in seven cell lines representative of major mamalian body systems and derived from two different animals species and humans.
INNOTARGETS has established a joint training syllabus designed to widen the career prospects of the doctoral students, and to optimize their professional competences. During the training period, technical courses (transposom techniques and proteomes) as well as complementary skills course (data management, scientific writing, IPR rights, entrepreneurship and personal impact) have been offered to the network’s students. INNOTARGETS has communicated preliminary results to the scientific community through participation in international and national meetings and have contributed to public science events centered on the topic of the global antimicrobial crises.
The INNOTARGETS project is expected to generate concrete impact for the beneficiaries in terms of generation of knowledge beyond the state of the art within bacterial metabolism during infection, metabolic responses to treatment with antimicrobials in antibiotic resistant bacteria, and on use of modelling for identification of suitable targets for antimicrobials and for toxicity prediction. Furthermore, the research is expected to lead to identification of putative targets for novel antimicrobials and helper drugs to resensitize resistant bacteria and block spread of resistance, lead molecules which blocks such targets, mathematical models of the metabolism in selected bacteria, software for toxicity prediction and improved screening platforms for antimicrobials, all of which will be valuable contribution to the global fight against antimicrobial resistance and the lack of novel antimicrobials.
The doctoral students of the network will gain strong skills that will significantly increase their market value and career prospects in academia and/or industry.
The doctoral students of the network will gain strong skills that will significantly increase their market value and career prospects in academia and/or industry.