Aptâmeros para o bloqueio de toxinas tipo shiga de Escherichia coli

The proposed project aims to develop an alternative therapeutic tool for dealing with Shiga toxin related disease, caused by STEC. This bacterium poses serious risks to public health worldwide due to their high prevalence and transmission through contaminated food. Severe infection cases have limited therapeutic options and very often rely on merely supportive procedures. The focus on developing aptamers as a therapeutic alternative thus aims to take advantage of the known advantages of these molecules, in recognizing and binding with high affinity to target molecules, to combat one of the main causes of food-borne diseases in humans. But this proposal also aims to go further. Traditional DNA/RNA aptamers have some limitations, mainly because natural nucleic acids are susceptibility to enzymatic degradation, which has slowed down their application in vivo. So, this proposal aims to advance aptamers development by: -Improving traditional aptamer with nucleic acid mimics (NAMs), modified nucleotides with unique thermal, chemical, and biological properties. This approach enhances the capabilities of aptamers, increasing stability while boosting their affinity and specificity for the target, offering a more promising solution for blocking Stx under in vivo conditions. -Using advanced biophysical techniques, based on computational modelling, to understand the structural and functional dynamics of aptamer-toxin interactions. This detailed analysis will refine aptamer optimization, improving stability and providing a foundation for tailored in silico methodologies in assisting aptamer development. The proposal aims to release a user-friendly interface that can be used by other researchers to predict/model NAM-aptamers structure/docking. Of notice, we have published the only functional pipeline for NAM-aptamers prediction; and we expect to take it to the next development/integration stage. -Focusing on the translation of aptamer-based therapeutics to in vivo situations through pre-clinical efficacy studies in cell lines and mouse models. These studies will demonstrate the ability of NAM aptamers to accelerate the translation process, showing the way forward for other future projects aimed at developing aptamer-based therapies for human use. In summary, the proposed project sets ambitious objectives aimed at transforming the therapeutic landscape for Shiga toxin-related infections. By integrating innovative molecules to enhance aptamer performance, elucidating aptamer-toxin interactions, testing their efficacy in vitro/vivo, and facilitating clinical translation; the project seeks to surpass the current state of the art in aptamer research. These efforts hold the potential to provide solutions that can latter on evolve to clinical trial and have a significant impact on patient outcomes.