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Design of ModProtease through High-Throughput Screening Technologies for Therapeutic Applications

Project description

Targeting hard-to-treat KRAS-driven cancers

Mutations of the KRAS gene result in some of the most aggressive cancers by hijacking cell growth signals, causing tumours to grow uncontrollably. These mutations are difficult to target because they act inside cells and often develop resistance to treatments. Supported by the Marie Skłodowska-Curie Actions programme, the ModProtease project aims to engineer protease-antibody combinations that precisely cut cancer-specific proteins to improve treatment effectiveness, while reducing side effects. Using a screening platform with glowing microbeads, researchers will identify the most promising protease variants. A clever molecular ‘tag-and-catch’ system links these proteases to antibodies, creating powerful targeted therapies. ModProtease offers new hope against KRAS-driven cancers.

Objective

The integration of ModProtease with antibody-based systems represents a promising strategy for targeted cancer therapy, particularly in addressing the challenges posed by KRAS mutations. These mutations are notoriously difficult to target directly with antibodies due to their involvement in intracellular signalling pathways and the development of resistance mechanisms. Strategies to overcome these limitations could include the development of protease-antibody conjugates and these conjugates might be directed to cleave specific substrates within tumour microenvironment, thereby enhancing the therapeutic effect of the antibody while minimizing off-target effects. However, engineered proteases remain underrepresented amongst protein-based biopharmaceuticals. This situation is due to fundamental challenges that exist in redirecting the substrate specificity of proteases. We propose a high-throughput screening platform that employs monoclonal hydrogel microbeads containing target substrates with FRET-based detection systems allowing for the precise identification of protease activity. This enables the rapid evaluation of various ModProtease variants and their effectiveness when conjugated to antibodies. The interplay of the SpyCatcher-SpyTag system enables the efficient and specific conjugation of ModProteases to antibodies, facilitating the creation of potent antibody-protease. Following high-throughput screening using fluorescence-activated bead sorting (FABS), the platform facilitates the recovery and analysis of protease genotypes from the microbeads. The group of Prof. Hollfelder (University of Cambridge) has substantial expertise in high-throughput methods and regarding the biology of proteases and protein binders. These conditions make the Host laboratory ideal for this project and will provide critical insights and technical support. This project will also collaborate with AstraZeneca, and targeted proteolysis against cancer cells will be assessed.

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HORIZON-TMA-MSCA-PF-EF - HORIZON TMA MSCA Postdoctoral Fellowships - European Fellowships

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Call for proposal

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(opens in new window) HORIZON-MSCA-2024-PF-01

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Coordinator

THE CHANCELLOR MASTERS AND SCHOLARS OF THE UNIVERSITY OF CAMBRIDGE
Net EU contribution

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€ 260 347,92
Address
TRINITY LANE THE OLD SCHOOLS
CB2 1TN CAMBRIDGE
United Kingdom

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Region
East of England East Anglia Cambridgeshire CC
Activity type
Higher or Secondary Education Establishments
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Total cost

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