Periodic Reporting for period 1 - CypTox (Training next level scientists and researchers to develop highly selective and safe insecticides)
Reporting period: 2021-01-01 to 2022-12-31
Some of the most acute challenges that the world faces now and in the foreseeable future are caused by insects and mites that seriously threaten human health and food security. Their control is primarily achieved by the use of insecticides: for example, malaria prevalence has halved since 2000, saving 660 million lives, with 80% of the reduction being attributable to the use of insecticides. However, both the limited availability of low risk insecticides and insecticide resistance represent a major threat, and there is an urgent need to develop new insecticides. CypTox will apply biotechnology excellence to exploit the cytochrome P450 (CYP) metabolic/detoxification pathway of target and non-target organisms, to develop novel insecticides, efficient against selected insect & mite major pests and vectors, but are highly selective and safe for mammals, pollinators and the environment. Outcomes will also include biotechnology-based platforms (high-throughput cell/enzyme- screening assays and in silico pipelines), in line and beyond Pharma state of the art, that will advance research capabilities, for future developments of low risk insecticides. CypTox will provide excellent research training within a creative and flexible environment that actively promotes the integration of academic rigor and commercial pragmatism through mobility between sectors and focused training events. The size and balance of a highly motivated consortium (6 academics/6 industrial in EU and 3 TC partners; 300 well distributed secondments), the involvement of experienced PIs in Horizon 2020 and ERC projects and the modern communication, dissemination and exploitation approaches of CypTox will ensure efficient implementation and impact, at several levels. Enhanced career perspectives for scientists and researchers will be achieved by building on a sustainable multidisciplinary and inter-sectoral network in a wide range of fields, with the integration of world leading researchers and stakeholders.
The overarching objective of CypTox is to exploit the primary P450-based metabolic pathway of target and non-target organisms, for the development of highly selective, safe insecticides and new resistance breaking formulations, to improve the efficiency and sustainability of pests and vector control.
The overarching objective of CypTox is to exploit the primary P450-based metabolic pathway of target and non-target organisms, for the development of highly selective, safe insecticides and new resistance breaking formulations, to improve the efficiency and sustainability of pests and vector control.
The following work has been carried out by the consortium during the CypTox project’s first 24 months towards the achievement of the specific project objectives:
Objective 1. To coordinate and manage all aspects of the work.
• Definitive work plan accepted and project started. Consortium/Partnership Agreement singed.
• Launch of CypTox website (www.cyptox.eu).
• Organization of the CypTox Kick-off meeting and the Mid-Term Meeting.
• 14 Deliverables submitted; 3 milestones achieved.
• 2 articles in scientific peer reviewed journals published.
• Several communication and dissemination activities (workshops, open day events-public outreach activities, synergies with other projects etc) took place.
Objective 2. To curate in depth cytochrome P450s sequences and benchmark gene expression associated with xenobiotic metabolism and selective toxicity.
• Phylogenetic analysis completed
• Contribution of CYPs in xenobiotic metabolism, insecticide resistance and specificity were studied.
• Important correlations between phenotypic resistance and CYP expression profiles were revealed.
• A database of CYP sequences and expression levels was generated.
Objective 3. To develop robust, reproducible and scalable P450-based screening tools and assays (R&I) for in vivo and in vitro screens of novel insecticides and synergists.
• Optimisation of conditions for CYP expression in bacteria and subsequent progress towards scale up was performed.
• Functional expression of arthropodal P450s in bacteria was successfully completed.
• High throughput assays were developed and applied for CYPs.
• Stably transformed insect cell lines: implementation plan was formed.
Objective 4. To design, optimise and in vitro test synergists and pro-insecticides that strongly interact with selective and resistance conferring CYPs, as well as transfer specialised knowledge.
• Key interactions between important pest CYPs and relevant chemical compounds / leads, were studied
• Molecular dynamics simulations were proven to allow a better understanding of motions associated to ligand binding, which are crucial for design.
• Catalytic activity of recombinantly expressed enzymes/microsomes was measured.
Objective 5. To validate the efficacy of novel compounds, and some additional available green chemistry compounds, against target pest and as well as monitor effects on non-target organisms.
• Preparatory work performed to establish Controlled environment (CE) bioassays.
• Setup of laboratory bioassays with synergists has started.
• Protocols were developed for the cell testing procedure using different assays.
Objective 6 To promote training towards the Personal Career Development Plan for each researcher and scientist, and integrate knowledge for modern biotechnology tools, and the value and rational use of safe insecticides, for the efficient and sustainable pest and disease control.
• Workshop: “Personal Career Development Plan” was organized.
• Practical Course: “Enzyme Biotechnology applications for Pesticide/Drug Discovery” was organized.
• Additional training activities in the local host institutes during secondments took place.
• Synergies with other relevant projects were established.
• The future implementation plans for training, networking and knowledge integration was formed.
• CypTox Innovation Management and Commercialisation Committee (IMCC) established.
Objective 1. To coordinate and manage all aspects of the work.
• Definitive work plan accepted and project started. Consortium/Partnership Agreement singed.
• Launch of CypTox website (www.cyptox.eu).
• Organization of the CypTox Kick-off meeting and the Mid-Term Meeting.
• 14 Deliverables submitted; 3 milestones achieved.
• 2 articles in scientific peer reviewed journals published.
• Several communication and dissemination activities (workshops, open day events-public outreach activities, synergies with other projects etc) took place.
Objective 2. To curate in depth cytochrome P450s sequences and benchmark gene expression associated with xenobiotic metabolism and selective toxicity.
• Phylogenetic analysis completed
• Contribution of CYPs in xenobiotic metabolism, insecticide resistance and specificity were studied.
• Important correlations between phenotypic resistance and CYP expression profiles were revealed.
• A database of CYP sequences and expression levels was generated.
Objective 3. To develop robust, reproducible and scalable P450-based screening tools and assays (R&I) for in vivo and in vitro screens of novel insecticides and synergists.
• Optimisation of conditions for CYP expression in bacteria and subsequent progress towards scale up was performed.
• Functional expression of arthropodal P450s in bacteria was successfully completed.
• High throughput assays were developed and applied for CYPs.
• Stably transformed insect cell lines: implementation plan was formed.
Objective 4. To design, optimise and in vitro test synergists and pro-insecticides that strongly interact with selective and resistance conferring CYPs, as well as transfer specialised knowledge.
• Key interactions between important pest CYPs and relevant chemical compounds / leads, were studied
• Molecular dynamics simulations were proven to allow a better understanding of motions associated to ligand binding, which are crucial for design.
• Catalytic activity of recombinantly expressed enzymes/microsomes was measured.
Objective 5. To validate the efficacy of novel compounds, and some additional available green chemistry compounds, against target pest and as well as monitor effects on non-target organisms.
• Preparatory work performed to establish Controlled environment (CE) bioassays.
• Setup of laboratory bioassays with synergists has started.
• Protocols were developed for the cell testing procedure using different assays.
Objective 6 To promote training towards the Personal Career Development Plan for each researcher and scientist, and integrate knowledge for modern biotechnology tools, and the value and rational use of safe insecticides, for the efficient and sustainable pest and disease control.
• Workshop: “Personal Career Development Plan” was organized.
• Practical Course: “Enzyme Biotechnology applications for Pesticide/Drug Discovery” was organized.
• Additional training activities in the local host institutes during secondments took place.
• Synergies with other relevant projects were established.
• The future implementation plans for training, networking and knowledge integration was formed.
• CypTox Innovation Management and Commercialisation Committee (IMCC) established.
The comprehensive molecular and functional study of the CYPome of smart selection of target and non-target organisms, for the development by design of highly selective, safe insecticides and new resistance breaking formulations, will take this line of research to leap beyond the current state of the art. CypTox will close gaps between academic and industrial participants, improving tools and delivering products, in order to achieve high impact. It provides outstanding innovative aspects at the research level, which are will lead, among others to novel robust selectivity evaluation pipelined and gene targets, pro-drug/anti-resistance insecticides leads, an expanded portfolio of low risk/green chemicals, as well as relevant targeted training activities.
The technologies that we develop are designed to be modular in the sense that they are not restricted to just the project-specific pests and vectors, but can be readily adapted to targets with similar profiles, and thus have a high potential for replication and wider impact. This also facilitates broad market uptake and creates strong financial incentives.
The outcome of reduced pesticide use can have significant and clear societal impacts. The main societal benefits of this project include benefits to human health and the production of safe insecticides, reduction of environmental pollution, and the increased sustainability of agriculture and public health. Significant environmental benefits of reduced synthetic pesticide use are also include less use of potentially polluting materials, less risk of off-target effects, increased biodiversity, and reduced risk of contamination of water streams/sources to provide just a few examples.
The technologies that we develop are designed to be modular in the sense that they are not restricted to just the project-specific pests and vectors, but can be readily adapted to targets with similar profiles, and thus have a high potential for replication and wider impact. This also facilitates broad market uptake and creates strong financial incentives.
The outcome of reduced pesticide use can have significant and clear societal impacts. The main societal benefits of this project include benefits to human health and the production of safe insecticides, reduction of environmental pollution, and the increased sustainability of agriculture and public health. Significant environmental benefits of reduced synthetic pesticide use are also include less use of potentially polluting materials, less risk of off-target effects, increased biodiversity, and reduced risk of contamination of water streams/sources to provide just a few examples.