Examining the modality-dependent target specificity of bioPROTACs by analysing global proteome changes following mRNA-encoded or recombinant protein biodegrader delivery.
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ABSTRACT: Proteolysis targeting chimeras (PROTACs) have surged in popularity as a drug modality owing to their ability to degrade pathogenic proteins. Despite their advantages over conventional inhibitors, PROTAC discovery still requires high-affinity warheads, but ligands for undruggable proteins are difficult to develop. As an alternative, protein based biological PROTACs (bioPROTACs) have been explored as a potential therapeutic. Since their substrate affinity is conferred by small-protein scaffolds, bioPROTACs are more easily developed for undruggable targets. Currently, bioPROTAC delivery is accomplished via electroporation, microinjection, or DNA/mRNA transfection, but these methods are either poorly translated to the clinic or suffer from manufacturing disadvantages. We developed a method to encapsulate and deliver recombinant bioPROTACs intracellularly using lipid nanoparticles. This study aims at evaluating differential degradation patterns between the same bioPROTAC delivered either as protein or as mRNA. To accomplish this, Ras-targeting bioPROTAC protein or mRNA were delivered to HEK293T cells or 8 hours. On- and off- target degradation was studied by LC-MS/MS shotgun proteomics. A label free based quantitative approach was used to identify significantly altered proteins in either treatment group. Gene ontology (GO) biological process and molecular function analyses were performed on significantly degraded proteins to determine target specificity for protein and mRNA bioPROTAC modalities.
INSTRUMENT(S): LTQ Orbitrap
ORGANISM(S): Homo Sapiens (human)
TISSUE(S): Cell Culture, Early Embryonic Cell
SUBMITTER: Andrew Tsourkas
LAB HEAD: Andrew Tsourkas
PROVIDER: PXD049388 | Pride | 2024-06-27
REPOSITORIES: Pride
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