Multi-chain chimeric antigen receptor and uses thereof
The present invention relates to a new generation of chimeric antigen receptors (CAR) referred to as multi-chain CARs. Such CARs, which aim to redirect immune cell specificity and reactivity toward a selected target exploiting the ligand-binding domain properties, comprise separate extracellular ligand binding and signaling domains in different transmembrane polypeptides. 91The signaling domains are designed to assemble in juxtamembrane position, which forms flexible architecture closer to natural receptors, that confers optimal signal transduction. The invention encompasses the polynucleotides, vectors encoding said multi-chain CAR and the isolated cells expressing them at their surface, in particularly for their use in immunotherapy. The invention opens the way to efficient adoptive immunotherapy strategies for treating cancer and viral infections.
1. A multi-chain Chimeric Antigen Receptor (CAR) including at least two different and separate transmembrane polypeptides expressed on an immune cell, comprising:
a first transmembrane polypeptide fused with an extracellular ligand-binding domain, the extracellular ligand-binding domain comprising a single-chain antibody fragment (scFv) that recognizes an antigen on a target cell, wherein the first transmembrane polypeptide comprises an alpha chain of Fc epsilon Receptor I (FcεRI) without its high affinity IgE binding domain, and
at least a second transmembrane polypeptide comprising at least one intracellular signal transducing domain from a T cell Receptor (TCR) zeta chain;
wherein the first transmembrane polypeptide dimerizes with the second transmembrane polypeptide to form the multi-chain CAR on the immune cell.
2. The multi-chain CAR of claim 1 , wherein the second transmembrane polypeptide lacks an extracellular ligand-binding domain.
3. The multi-chain CAR of claim 2 , further comprising a costimulatory domain of CD28.
4. The multi-chain CAR of claim 2 , further comprising a costimulatory domain of CD137.
5. The multi-chain CAR of claim 1 , wherein said first transmembrane polypeptide further comprises a CD8 stalk region.
6. The multi-chain CAR of claim 2 , wherein said first transmembrane polypeptide further comprises a CD8 stalk region.