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Projeto de investigação
Exploring one-hit-multiple-target strategies for Lung Cancer
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Publicações
Molecular Beacon for Detection miRNA-21 as a Biomarker of Lung Cancer
Publication . Alexandre, Daniela; Teixeira, Bernardo; Rico, André; Valente, Salete; Craveiro, Ana; Baptista, Pedro V.; Cruz, Carla; DCV - Departamento de Ciências da Vida; UCIBIO - Applied Molecular Biosciences Unit; MDPI - Multidisciplinary Digital Publishing Institute
Lung cancer (LC) is the leading cause of cancer-related death worldwide. Although the diagnosis and treatment of non-small cell lung cancer (NSCLC), which accounts for approximately 80% of LC cases, have greatly improved in the past decade, there is still an urgent need to find more sensitive and specific screening methods. Recently, new molecular biomarkers are emerging as potential non-invasive diagnostic agents to screen NSCLC, including multiple microRNAs (miRNAs) that show an unusual expression profile. Moreover, peripheral blood mononuclear cells’ (PBMCs) miRNA profile could be linked with NSCLC and used for diagnosis. We developed a molecular beacon (MB)-based miRNA detection strategy for NSCLC. Following PBMCs isolation and screening of the expression profile of a panel of miRNA by RT-qPCR, we designed a MB targeting of up-regulated miR-21-5p. This MB 21-5p was characterized by FRET-melting, CD, NMR and native PAGE, allowing the optimization of an in-situ approach involving miR-21-5p detection in PBMCs via MB. Data show the developed MB approach potential for miR-21-5p detection in PBMCs from clinical samples towards NSCLC.
Cellular modulation of a G-quadruplex structure found in the lung cancer-related microRNA-3196
Publication . Alexandre, Daniela; Polido, Joana; Miranda, André; Hudson, Robert H. E.; Monchaud, David; Baptista, Pedro V.; Cruz, Carla; UCIBIO - Applied Molecular Biosciences Unit; DCV - Departamento de Ciências da Vida; Elsevier
RNA G-quadruplexes (G4s) are promising drug targets due to their high cellular abundance. G-rich RNA regions inherently form G4 structures, while GC-rich sequences adopt stem-loop conformations, and their dynamic equilibrium critically influences RNA function. MicroRNAs (miRs), key regulators of protein expression, undergo processing by Dicer, which specifically recognizes stem-loop structures in precursor miRs (pre-miRs). Notably, some pre-miRs containing G4-forming sequences influence Dicer cleavage, suggesting that G4s can directly regulate miR production. Moreover, pre-miRs with G4 structures present promising targets for small molecules. This research focuses on identifying and modulating G4 structure in pre-miR-3196 to restore normal lung cancer (LC) levels, offering a potential therapeutic strategy. Firstly, bioinformatic analyses indicated the presence of G4 motifs in pre-miR-3196. We then demonstrated in vitro that this RNA sequence folds into stable G4s by a combination of biophysical and biochemical assays. Then, we demonstrated the formation of these G4s in human cancer cells by confocal imaging before showing that these G4s can be modulated using the RNA G4 destabilizer PhpC, which impacts the miR-3196 biogenesis. These findings highlighted the possibility of using G4s to control the expression of mature miR-3196 and revealed the potential of using the destabilizer PhpC to adjust its G4 structure.
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Entidade financiadora
Fundação para a Ciência e a Tecnologia
Programa de financiamento
POR_CENTRO
Número da atribuição
2021.07695.BD