Actividad Investigadora
Development of an antibody for immunoaffinity profiling of NEDDylated proteins in human pathologies
Datos básicos
- Código del Financiador:
- INNVAL19/19
- Año Inicial:
- 2020
- Año final:
- 2022
Objetivos del proyecto
The fact that the function of proteins is affected not solely by the message contained in the genome but by many other factors, has put the stress in the role of postranslational modifications in human pathology. This is specially relevant to pathologies without a clear relationship between the mutations found and their etiology. However, the study of postranslational modifications require the use of specific tools that could be used to analyze their profile and dynamics in different pathologies and therapeutical contexts. This is the case of NEDDylation, a modification that presents many similarities to ubiquitination, but that has recently taken relevance due to the promising results obtained in different types of cancers with the drug MLN4924-Pevonedistat, that specifically inhibits this modification. NEDDylation is involved in the stability, subcellular localization and function of many proteins, modulating processes like DNA repair, chromatin remodeling, signaling transduction and gene expression. Its role in several hematological and solid malignancies as well as in neurodegenerative diseases has been supported by different observations and MLN4924 is being tested in phase II clinical trials. The current analyses of ubiquitination and other ubiquitin-like modifications in different pathologies use antibodies that recognize small polyaminoacidic remnants left over the modified residues after digestion with trypsin. These antibodies are proprietary of certain companies that offer postranslational modification profiling services. Although this method represents a breakthrough in the study of postranslational modifications at a whole cell level, the antibody used to date lacks the specificity to discriminate NEDDylation from ubiquitination and ISGylation. The aim of the present project is to develop a monoclonal antibody that could be used to specifically detect NEDDylation in pathology-profiling assays. Taking advance of a longer and differential aminoacidic remnant left on NEDDylated lysines after digestion with the endopeptidase Lys-C, we will rise a specific antibody against it that could discriminate them from ubiquitin-, SUMO- or ISG15-modified lysines. The advantage of this antibody in investigational and clinical applications is clear: it will allow to determine the extension of NEDDylation in candidate pathologies, identify the differentially modified proteins, design novel single or combinational therapies that use MLN4924-Pevonedistat and follow their action over time and dose. Our aim is the protection of the monoclonal antibody for its commercial exploitation, either locally or by its transfer to a company with postranslational modification profiling services.
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