Method and compositions for cellular immunotherapy
The present invention provides nucleic acids, vectors, host cells, methods and compositions to confer and/or augment immune responses mediated by cellular immunotherapy, such as by adoptively transferring CD8+ central memory T cells or combinations of central memory T cells with CD4+ T cells that are genetically modified to express a chimeric receptor. In embodiments the genetically modified host cell comprises a nucleic acid comprising a polynucleotide coding for a ligand binding domain, a polynucleotide comprising a customized spacer region, a polynucleotide comprising a transmembrane domain, and a polynucleotide comprising an intracellular signaling domain. It has been surprisingly found that the length of the spacer region can affects the ability of chimeric receptor modified T cells to recognize target cells in vitro and affects in vivo efficacy of the chimeric receptor modified T cells. Pharmaceutical formulations produced by the method, and methods of using the same, are also described.
1. A chimeric receptor, the chimeric receptor comprising an extracellular binding region consisting of an scFv and a polypeptide spacer, a transmembrane domain and an intracellular signaling domain, wherein:
(a) the scFv binds to an epitope in the extracellular region of CD19, wherein the scFv comprises a variable heavy (VH) chain domain comprising the sequences DYGVS (SEQ ID NO:74), VIWGSETTYYNSALKS (SEQ ID NO:60) and YAMDYWG (SEQ ID NO:61), and a variable light (VL) chain domain comprising the sequences RASQDISKYLN (SEQ ID NO:57), SRLHSGV (SEQ ID NO:58) and GNTLPYTFG (SEQ ID NO:59);
(b) the polypeptide spacer is located between the scFv and the transmembrane domain, wherein the polypeptide spacer consists of the amino acid sequence of SEQ ID NO:21; and
(c) the intracellular signaling domain comprises the signaling domain of CD3ζ and a costimulatory domain.
2. A chimeric receptor, the chimeric receptor comprising:
(a) an scFv that binds to an epitope in the extracellular region of CD19, wherein the scFv comprises the variable heavy (VH) chain domain and the variable light (VL) chain domain encoded by SEQ ID NO:3;
(b) a transmembrane domain;
(c) a polypeptide spacer located between the scFv and the transmembrane domain, wherein the polypeptide spacer consists of the amino acid sequence of SEQ ID NO:21; and
(d) an intracellular signaling domain that comprises the signaling domain of CD3ζ and a costimulatory domain.
3. The chimeric receptor of claim 2 , wherein the scFv is encoded by SEQ ID NO:3.
4. The chimeric receptor of claim 1 , wherein:
the transmembrane domain consists of the transmembrane domain of CD8 or of CD28; and
the costimulatory domain comprises the signaling domain of 4-1BB or of CD28.
5. The chimeric receptor of claim 2 , wherein:
the transmembrane domain consists of the transmembrane domain of CD8 or of CD28; and
the costimulatory domain comprises the signaling domain of 4-1BB or of CD28.
6. The chimeric receptor of claim 1 , wherein:
the transmembrane domain consists of the amino acid sequence encoded by the polynucleotide sequence of SEQ ID NO:5; and
the intracellular signaling domain comprises the amino acid sequence encoded by the polynucleotide sequence of SEQ ID NO:6 and the amino acid sequence encoded by SEQ ID NO:7.
7. The chimeric receptor of claim 2 , wherein:
the transmembrane domain consists of the amino acid sequence encoded by the polynucleotide sequence of SEQ ID NO:5; and
the intracellular signaling domain comprises the amino acid sequence encoded by the polynucleotide sequence of SEQ ID NO:6 and the amino acid sequence encoded by SEQ ID NO:7.
8. The chimeric receptor of claim 1 , wherein the chimeric receptor comprises:
the anti-CD19 scFv encoded by SEQ ID NO:3;
the polypeptide spacer encoded by SEQ ID NO:4;
the transmembrane domain encoded by SEQ ID NO:5;
the costimulatory domain encoded by SEQ ID NO:6; and
the CD3ζ signaling domain encoded by SEQ ID NO:7.
9. A T cell comprising the chimeric receptor of claim 1 .
10. A composition, comprising the T cell of claim 9 in a pharmaceutically acceptable excipient.
11. A T cell comprising the chimeric receptor of claim 8 .
12. A composition, comprising the T cell of claim 11 in a pharmaceutically acceptable excipient.
13. A T cell comprising the chimeric receptor of claim 2 .
14. A composition, comprising the T cell of claim 13 in a pharmaceutically acceptable excipient.
15. A T cell comprising the chimeric receptor of claim 6 .
16. A composition, comprising the T cell of claim 15 in a pharmaceutically acceptable excipient.
17. A T cell comprising the chimeric receptor of claim 7 .
18. A composition, comprising the T cell of claim 17 in a pharmaceutically acceptable excipient.