| Abstract | Insecticides remain indispensable for managing insect populations that transmit life-threatening diseases such as malaria and dengue fever. However, the rising evolution of insecticide resistance in mosquitoes poses a significant threat to global health initiatives. Most conventional insecticides target the insect nervous system, using neurotoxic agents that disrupt sodium channels (e.g., DDT and pyrethroids) or interfere with cholinergic signalling (e.g., organophosphates, carbamates, and neonicotinoids). Over time, mosquito populations have developed resistance to these chemicals, reducing their effectiveness. Overcoming this challenge calls for proactive resistance management strategies and the development of insecticides with novel modes of action. However, progress has been slowed by the labor-intensive and time-consuming nature of traditional screening methods, such as larval and adult bioassays, which limit high-throughput discovery. Efforts to repurpose existing chemicals, like herbicides, for insecticidal purposes have also faced difficulties, particularly in identifying molecular targets effective against mosquitoes. These challenges are compounded by a limited understanding of resistance and cross-resistance mechanisms, further complicating the discovery of effective new insecticides. In synergy with LSTM's Insecticide Resistance Prediction Platform (IRPP; https://www.lstmed.ac.uk/irpp ) and building on previous research utilising chemical proteomics to pre-emptively identify resistance markers in mosquitoes, this project seeks to deepen the understanding of molecular mechanisms behind resistance and cross-resistance, while providing a tool to speed up the discovery of new insecticides. The research will focus on establishing mosquito cell lines capable of differentiating into neuron-like cells to study insecticide resistance at the molecular level and identify their insecticidal targets. Additionally, it will involve developing a high-throughput platform that uses electrophysiological recordings to assess the effects of insecticides on mosquito neuron function. The project will also screen for new insecticides that target alternative pathways, focusing on non-neurotoxic mechanisms of action. The in vitro findings will be validated using in vivo models to ensure accurate predictions of insecticide efficacy and accelerate the identification of effective insecticide candidates. |
| Where does this project lie in the translational pathway? | T1 - Basic Research |
| Expected Outputs | 1.Establish a new mosquito cell line platform capable of differentiating into neuron-like cells, providing a vital resource for studying insecticide resistance and facilitating the development of high-throughput screening tools for insecticide discovery. 2.Develop cell culture-based assays for high-throughput functional validation of genetic variants, accelerating the identification of therapeutic targets and resistance mechanisms in disease-carrying vectors. 3.Identify patterns of cross-resistance and discover new insecticidal targets to combat emerging resistance mechanisms in vector populations. 4.Develop non-invasive techniques to monitor neural activity in mosquitoes, enhancing the study of neurological responses in this model. 5.Publish high-impact research in leading journals, advancing the fields of vector biology, neurobiology, and insecticide resistance. 6.Develop expertise in quantitative bioinformatics analysis, focusing on the integration and interpretation of large-scale datasets across multiple biological disciplines. |
| Training Opportunities | 1. Gain expertise in protein expression analysis using various proteomic techniques and mass spectrometry. 2. Develop skills in transcriptomics and bioinformatics to understand gene expression and resistance mechanisms. 3. Learn high throughput screening techniques, including electrophysiology assays for both in vitro and in vivo models. 4. Build laboratory skills in molecular biology and cell culture techniques. 5. Gain proficiency in generating, analyzing, and integrating large-scale ‘omics datasets. 6. Acquire hands-on experience in vector control strategies and insecticide resistance research. 7. Enhance presentation skills by showcasing research at national and international conferences. And receive mentorship in writing and publishing high-impact scientific papers. |
| Skills Required | Background in molecular biology, neurobiology, or pharmacology. Hands-on experience with electrophysiology, RNA sequencing (RNA-seq), and protein analysis (deserable but not essential) Keen interest in bioinformatics and data analysis, with a focus on interpreting complex biological data. |
|
Key Publications associated with this project |
Moyes, C. L., Lees, R. S., Yunta, C., Walker, K. J., Hemmings, K., Oladepo, F., Hancock, P. A., Weetman, D., Paine, M., & Ismail, H. M. (2021). Assessing cross-resistance within the pyrethroids in terms of their interactions with key cytochrome P450 enzymes and resistance in vector populations. Parasites & vectors, 14(1), 115. https://doi.org/10.1186/s13071-021-04609-5 |
| Ismail, H. M.; O’Neill, P. M.; Hong, D. W.; Finn, R. D.; Henderson, C. J.; Wright, A. T.; Cravatt, B. F.; Hemingway, J.; Paine, M. J., Pyrethroid activity-based probes for profiling cytochrome P450 activities associated with insecticide interactions. Proceedings of the National Academy of Sciences 2013, 110, 19766-19771. | |
| Riveron, J. M.; Yunta, C.; Ibrahim, S. S.; Djouaka, R.; Irving, H.; Menze, B. D.; Ismail, H. M.; Hemingway, J.; Ranson, H.; Albert, A., A single mutation in the GSTe2 gene allows tracking of metabolically based insecticide resistance in a major malaria vector. Genome Biol 2014, 15, R27. | |
| David, J. -P.; Ismail, H. M.; Chandor-Proust, A.; Paine, M. J. I., Role of cytochrome P450s in insecticide resistance: impact on the control of mosquito-borne diseases and use of insecticides on Earth. Philosophical Transactions of the Royal Society of London B: Biological Sciences 2013, 368, 20120429. | |
| Gaburro, J., Duchemin, JB., Paradkar, P.N. et al. Electrophysiological evidence of RML12 mosquito cell line towards neuronal differentiation by 20-hydroxyecdysdone. Sci Rep 8, 10109 (2018). https://doi.org/10.1038/s41598-018-28357-2 |