Descrizione del progetto
Approfondire il ruolo complesso delle proteine leganti l’RNA nella stabilità dell’RNA stesso
Le cellule T CD8+ sono fondamentali per il controllo delle infezioni e l’eliminazione delle cellule tumorali tramite citochine infiammatorie e molecole citotossiche. La produzione di queste molecole effettrici richiede una regolazione per prevenirne il malfunzionamento, che può provocare malattie potenzialmente letali. La regolazione della stabilità dell’RNA e dell’efficienza di traslazione da parte di numerose proteine leganti l’RNA rimane poco conosciuta. Il progetto PRinTERs, finanziato dall’UE, utilizzerà la genetica del topo e la biologia molecolare e cellulare per studiare il controllo della produzione di citochine da parte delle proteine leganti l’RNA. Alcune tecniche di proteomica altamente sensibili individueranno il repertorio delle proteine leganti l’RNA nei linfociti T umani, mentre screening funzionali rileveranno le proteine che controllano effettori specifici. Le proteine leganti l’RNA selezionate saranno esaminate per il loro ruolo nella risposta dei linfociti T alle infezioni e alla formazione di tumori.
Obiettivo
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.
Campo scientifico
Programma(i)
Argomento(i)
Meccanismo di finanziamento
ERC-COG - Consolidator GrantIstituzione ospitante
1006 AN Amsterdam
Paesi Bassi