Exploring emerging biomarkers for the management of heart failure in DBS

• Autor(a) principal: Colaço, Beatriz dos Santos
• Data de Publicação: 2024
• Tipo de documento: Dissertação
• Idioma: eng
• Título da fonte: Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
• Texto Completo: http://hdl.handle.net/10773/44459
• topic: Heart failure; Proteomics; DBS; Biomarkers; Levosimendan; Bioinformatics

Introduction: Heart failure (HF) is a highly prevalent cardiovascular condition, characterized by the heart’s inability to pump blood adequately to meet the body’s metabolic needs. The detection of protein biomarkers has shown great potential for early detection and monitoring of HF, with significant implications for improving therapeutic approaches. In cases of advanced HF, levosimendan therapy, an inotropic agent that enhances cardiac contractility, has been used with the aim of temporarily improving cardiac function and, consequently, the quality of life of patients. Objectives: This study aims to validate the Dry Blood Spots (DBS) method for the analysis of protein biomarkers in patients with HF and to investigate the potential associations between levosimendan therapy and cardiac biomarkers. Methods: The DBS method was validated in a cohort of 26 patients with HF with reduced ejection fraction (<40%). For this purpose, the proteomic profile in DBS and plasma samples was compared using SDS-Page. The profile bands were digested and analyzed by mass spectrometry (MS). Subsequently, an in sílico analysis was conducted to predict potential targets of levosimendan. To validate the results obtained and to observe the impact of levosimendan on key HF biomarkers (MMP-9, TIMP-1, Gal-3, and BNP), a targeted proteomic analysis was performed via Slot Blot on DBS samples. The statistical analysis was supported by bioinformatics tools, which facilitated a better interpretation of the results in light of their biological significance. Results and Discussion: A total of 134 proteins were identified by MS, 55 of which were found to be common and similarly quantified between DBS and plasma samples, validating the use of DBS for proteomic analysis. MMP-9 stood out as one of the main targets for levosimendan. Additionally, the effect of levosimendan on the levels of MMP-9, TIMP-1, Gal-3, and BNP was studied through Slot Blot analysis. When comparing biomarker levels between patients undergoing levosimendan therapy and those not receiving it, a decrease in Gal-3 and MMP- 9 levels and an increase in TIMP-1 levels were observed in patients treated with levosimendan. These results reinforce the importance of these biomarkers in the monitoring of HF. Bioinformatic analysis corroborated that biomarkers such as Gal-3, TIMP-1, CRP, CysC, MMP-9, and BNP are associated with HF and hold potential for use in the diagnosis and monitoring of the disease. Conclusion: The results indicate that the proteomic analysis method using DBS is a viable alternative to plasma. Additionally, it was found that levosimendan may temporarily improve cardiac function, contributing to the quality of life of patients with advanced HF. However, it is important to validate these findings in a larger cohort to ensure their robustness. The association of MMP-9, TIMP-1, and Gal-3 with HF reinforces their role in the prognosis and monitoring of the disease, paving the way for new therapeutic approaches.