Development of metrics of quantitative proteomics capable of capturing the functional and compositional complexity of high-density lipoprotein

• Grant number: 23/00995-4
• Support Opportunities: Regular Research Grants
• Start date: October 01, 2023
• End date: September 30, 2025
• Field of knowledge: Biological Sciences - Biochemistry
• Principal Investigator: Graziella Eliza Ronsein
• Grantee: Graziella Eliza Ronsein
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: Thomas Quad de Aguiar Vallim

Abstract

Since pioneering clinical studies were performed in the 1970s, it is known that the levels of cholesterol (C) in high-density lipoprotein (HDL) correlate inversely with the chances of developing cardiovascular diseases. However, studies with drugs that increase the content of cholesterol in HDL (HDL-C, routinely measured in clinics) have had negative results, calling into check the importance of HDL. Today it is known that the term HDL defines a heterogeneous set of particles with complex lipid and protein composition, and that measurement of its cholesterol content is a simplification incapable of capturing its complexity, diversity and functionality. New metrics are urgently needed to understand the function of HDL particles. Regarding the set of proteins that make up HDL - its proteome - more than 500 proteins have been described as present in this particle, but there is no consistency and reproducibility among studies in the area. Our group pioneered the development of robust and reliable quantitative methodologies for the quantification of HDL proteome. From the application of the methods developed in the laboratory, we demonstrated a previously unknown role of HDL as a modulator of inflammatory and immune responses. Currently, there is no standardization regarding the methods of isolation of HDL particles from plasma, and the majority (>95%) of the HDL proteomic methods used in the literature are semi-quantitative and do not employ quality controls and/ or internal standards. In this view, the goal of this project is to develop an optimized workflow for isolation of HDL particles from plasma followed by quantification of their proteins, correlating the data obtained with clinical findings and with data obtained in functional assays, using cell culture of endothelial and immune cells. The results obtained may serve as a guideline to improve strategies for HDL isolation and proteome quantification, relieving the lack of consistency that hinders advances to understand the roles (known or not) of this lipoprotein. (AU)

Scientific publications

(6)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

GOMES, FRANCIELLE AGUIAR; SOUZA JR, DOUGLAS RICARDO; MASSAFERA, MARIANA PEREIRA; RONSEIN, GRAZIELLA ELIZA. Robust assessment of sample preparation protocols for proteomics of cells and tissues. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1872, n. 5, p. 13-pg., 2024-07-02. (23/00995-4, 13/07937-8, 16/00696-3)

GASPAR, RENATO S.; FRANCA, ALEFE ROGER SILVA; OLIVEIRA, PERCILLIA VICTORIA SANTOS; DINIZ-FILHO, JOEL FELIX SILVA; TEIXEIRA, LIVIA; VALADAO, IURI CORDEIRO; DEBBAS, VICTOR; SANTOS, CLENILTON COSTA DOS; MASSAFERA, MARIANA PEREIRA; BUSTOS, SILVINA ODETE; et al. Endothelial protein disulfide isomerase A1 enhances membrane stiffness and platelet-endothelium interaction in hyperglycemia via SLC3A2 and LAMC1. Journal of Thrombosis and Haemostasis, v. 22, n. 11, p. 17-pg., 2024-10-24. (20/15944-8, 13/07937-8, 22/05750-7, 23/00995-4, 20/03838-9, 16/00696-3)

ALEXANDRIA, GABRIELLY; VALERIO, HELLEN P.; MASSAFERA, MARIANA P.; REIS, LORENNA R.; COELHO, FERNANDO R.; DI MASCIO, PAOLO; RONSEIN, GRAZIELLA E.. The miniaturized isolation of neutrophil granules (MING) method allowed a deep proteome mapping of human neutrophil granules. Journal of Leukocyte Biology, v. N/A, p. 12-pg., 2024-12-05. (23/00995-4, 16/00696-3, 12/12663-1, 13/07937-8)

DEMPSEY, BIANCA; DA SILVA, BEATRIZ PEREIRA; CRUZ, LITIELE CEZAR; VILEIGAS, DANIELLE; SILVA, AMANDA R. M.; DA SILVA, RAILMARA PEREIRA; MEOTTI, FLAVIA CARLA. Unraveling the effects of uric acid on endothelial cells: A global proteomic study. REDOX BIOLOGY, v. 82, p. 21-pg., 2025-05-01. (13/07937-8, 18/14898-2, 12/12663-1, 23/00995-4)

REIS, LORENNA ROCHA; NASCIMENTO, RAFAELA OLIVEIRA; MASSAFERA, MARIANA PEREIRA; DI MASCIO, PAOLO; RONSEIN, GRAZIELLA ELIZA. Investigating neutrophil responses to stimuli: Comparative analysis of reactive species-dependent and independent mechanisms. REDOX BIOLOGY, v. 81, p. 11-pg., 2025-03-03. (12/12663-1, 13/07937-8, 16/00696-3, 23/00995-4)

DE FATIMA MELLO SANTANA, MONIQUE; SAWADA, MARIA ISABELA BLOISE ALVES CALDAS; SOUZA JUNIOR, DOUGLAS RICARDO; GIACAGLIA, MARCIA BENACCHIO; REIS, MOZANIA; XAVIER, JACIRA; CORREA-GIANNELLA, MARIA LUCIA; SORIANO, FRANCISCO GARCIA; GEBRIM, LUIZ HENRIQUE; RONSEIN, GRAZIELLA ELIZA; et al. Proteomic Profiling of HDL in Newly Diagnosed Breast Cancer Based on Tumor Molecular Classification and Clinical Stage of Disease. CELLS, v. 13, n. 16, p. 19-pg., 2024-08-01. (19/18431-4, 19/25702-4, 21/02401-9, 16/15603-0, 23/00995-4)