Using a novel protein degradation approach to uncover irf4-regulated genes in plasma cells
Plasma cells (PCs) are antibody-producing cells that develop from activated B cells in an immune response. Antibodies produced by these cells are essential for the clearance of pathogens and long-term protection against recurrent infections. PCs can also be pathogenic in autoimmune disease, where self-recognising antibodies are produced, and cancer, in the form of multiple myeloma. An understanding of the molecular pathways that control PC function is therefore necessary for understanding human immunity and these pathologies. Interferon Regulatory Factor 4 (IRF4, gene name Irf4) is a transcription factor whose deletion in mouse models results in the loss of all PCs. Because of this, the genes that are regulated by IRF4, and hence which cellular pathways are required for PC survival, are unknown. Here, I propose to use a revolutionary approach to determine the target genes of IRF4. I will use targeted protein degradation to deplete IRF4 in mouse PCs, and then determine the immediate changes in transcription following IRF4 loss using a state-of-the-art RNA sequencing technique, known as SLAM-seq. The advantage of this approach is that I can analyze changes in transcription before the onset of the survival defect. I will then study the functional roles of the identified IRF4 target genes in PCs. This is a multidisciplinary project that combines molecular biology approaches with cell biology and immune physiology. In addition, this proposal allows for transfer of knowledge from myself, an expert in the regulation of cell survival, and the host institution, which will train me in molecular biology techniques and expertise in PC biology. IRF4 has been implicated in both autoimmune disease and multiple myeloma, and so this proposal addresses a basic research question that has translatable outcomes. Hence, this proposal is in line with the H2020 objective to increase the transfer of knowledge into tangible products, and contributes to the European knowledge-based economy.