New molecular targets identification and validation of anti-inflammatory peptides applying distinct mass spectrometry-based proteomics approaches

Abstract

The aim of the CENTD project is to use animal venoms, secretions and venom-derived peptides as tools to identify and validate new molecular targets and signaling pathways of immunoinflammatory-related diseases to be useful for the discovery of new therapies and new drug development. For this purpose, our center has implemented and characterized distinct cellular models of inflammation (chondrocytes, macrophages, synoviocytes and neuron-like cells) and has identified five venom-derived peptides that presented anti-inflammatory and cytoprotection effects. The aging process of chondrocytes is associated with the degeneration of articular cartilage and is observed in rheumatoid arthritis and osteoarthritis. The treatment of these diseases lacks a specific or more powerful treatment and it is described that pro-inflammatory cytokines are key processes in the progression of these diseases. Therefore, finding new molecular targets to improve the treatment or to promote the development of new drugs for these inflammatory diseases is of great interest. Mass spectrometry based proteomics is versatile and a powerful analytical methodology that we aim to develop a workflow specific for new target discovery and target validation. For this purpose, our center has implemented mass spectrometry-based proteomics and has applied qualitative and relative quantitative label-free proteomics for identification of proteins and signaling pathways that were modulated on the distinct cellular models after treatment with peptides of interest. Moreover, in order to have an idea of peptide-protein interaction, our center has applied affinity-based profiling of protein cell extracts and peptides of interest and identified proteins from chondrocytes protein cellular extracts that interacted with the specific peptides. Therefore, we are now proposing two specific aims for this project: 1- to apply chemoproteomics as an activity-based profiling to identify the specific targets for the peptides of interest in chondrocytes and 2- to validate knockdown chondrocytes for specific targets using quantitative relative proteomics and targeted proteomics.