Projektbeschreibung
Untersuchung der komplexen Rolle von RNA-bindenden Proteinen bei der RNA-Stabilität
CD8+ T-Zellen sind für die Kontrolle von Infektionen und die Eliminierung von Tumorzellen durch entzündliche Zytokine und zytotoxische Moleküle unerlässlich. Die Produktion dieser Effektormoleküle muss reguliert werden, um ihre Fehlfunktion zu verhindern, die zu lebensbedrohlichen Erkrankungen führen kann. Die Regulierung der RNA-Stabilität und der Translationseffizienz durch zahlreiche RNA-bindende Proteine ist nach wie vor kaum verstanden. Im Rahmen des EU-finanzierten Projekts PRinTERs werden Mäusegenetik sowie Molekular- und Zellbiologie eingesetzt, um die Kontrolle der RNA-bindenden Proteine über die Zytokinproduktion zu untersuchen. Hochempfindliche Proteomik-Techniken werden das Repertoire RNA-bindender Proteine in menschlichen T-Zellen ermitteln, während funktionelle Screens diejenigen RNA-bindenden Proteine ermitteln werden, die spezifische Effektoren kontrollieren. Die ausgewählten RNA-bindenden Proteine werden auf ihre Rolle bei der Reaktion von T-Zellen auf Infektionen und bei der Tumorbildung untersucht.
Ziel
CD8+ T cells are critical to fight infections and to clear tumor cells through the production of inflammatory cytokines and cytotoxic molecules. These effector molecules must be tightly controlled: too little leads to the inability to control the pathogen, and too much can result in a life-threatening cytokine storm and tissue damage. While transcriptional control of effector genes is well-studied, regulation at the levels of RNA stability and translation efficiency by RNA-binding proteins (RBPs) has remained underappreciated. We recently found that several cytokines are tightly regulated through these processes, and we identified ZFP36L2 as one of the responsible RBPs. However, much is still to be learned about the underlying molecular mechanisms. Moreover, there are >1000 putative RBPs, and a systematic analysis of their regulatory activity in T cells is lacking, particularly with regard to the control of effector proteins.
Here, we will use a combination of mouse genetics, and molecular and cellular biology to gain a deep understanding of the control of cytokine production by RBPs, using ZFP36L2 as a paradigm. Next, we will take a novel, highly sensitive proteomics approach to systematically identify the RBP repertoire in resting and activated primary human T cells. Complementary functional screens will identify those RBPs that control specific effectors. Selected RBPs identified in these screens will be studied in-depth to understand their roles in T cell responses to acute infection and in tumor models. Lastly, we will define how RBPs can imprint and/or maintain the killer phenotype of human CD8+ T cells.
This research will significantly advance our understanding of post-transcriptional regulation of T cell effector activity, and it should help us to develop novel tools to drive effective T cell responses against pathogens and malignant cells.
Wissenschaftliches Gebiet
Programm/Programme
Thema/Themen
Finanzierungsplan
ERC-COG - Consolidator GrantGastgebende Einrichtung
1006 AN Amsterdam
Niederlande