IL-31 improves efficacy of macrophage-based adoptive cell therapy for cancer
The present invention provides macrophages genetically modified to express IL-31 or both IL-31 and a chimeric antigen receptor (CAR) for treatment of cancer. It further provides methods for treatment of cancer comprising administration of IL-31 along with genetically unmodified macrophages or genetically modified to express a CAR.
1. A macrophage which is genetically modified to express (i) human IL-31 of the sequence as set forth in SEQ ID No. 1 or a peptide which is at least 85% identical to the IL-31 sequence as set forth in SEQ ID No. 1 and which binds to IL-31 receptor alpha (IL-31RA); or (ii) a fused protein comprising human IL-31 of the sequence as set forth in SEQ ID No. 1 or a peptide which is at least 85% identical to the IL-31 sequence as set forth in SEQ ID No. 1 and which binds to IL-31RA, wherein in the fused protein the human IL-31 or peptide is attached to an immunoglobulin amino acid sequence comprising IgG.
2. The macrophages according to claim 1 , which is genetically modified to express human IL-31 of the sequence as set forth in SEQ ID No. 1.
3. A method of preparing genetically-modified macrophages according to claim 1 , comprising introducing a nucleic acid sequence encoding human IL-31, a peptide which is at least 85% homologous identical to the IL-31 sequence as set forth in SEQ ID No. 1 and which binds to IL-31RA, or the fused protein wherein the IL-31 or the peptide is attached to an immunoglobulin amino acid sequence comprising IgG, into a population of cells selected from monocytes, naïve macrophages or anti-tumor macrophages by transduction methods.
4. The method according to claim 3 , wherein the nucleic acid sequence encoding human IL-31 of the sequence as set forth in SEQ ID No. 1 is introduced into the cells by lentiviral transduction.
5. A pharmaceutical composition comprising genetically-modified macrophages according to claim 1 , and a pharmaceutically acceptable carrier.
6. The pharmaceutical composition according to claim 5 , wherein the macrophages are genetically modified to express human IL-31 of the sequence set forth in SEQ ID No. 1.
7. A method for treating cancer comprising administering to a patient in need thereof a therapeutically effective amount of the genetically modified macrophages according to claim 1 .
8. A method for treating cancer comprising administering to a patient in need thereof a therapeutically effective amount of the pharmaceutical composition according to claim 5 .
9. A macrophages which is genetically modified to express both: (i) human IL-31 of the sequence as set forth in SEQ ID No. 1 or peptide which is at least 85% identical to the IL-31 sequence as set forth in SEQ ID No. 1 and which binds to IL-31RA or a fused protein comprising human IL-31 of the sequence as set forth in SEQ ID No. 1 or peptide which is at least 85% identical to the IL-31 sequence as set forth in SEQ ID No. 1 and which binds IL-31RA, wherein in the fused protein the human IL-31 or the peptide is attached to an immunoglobulin amino acid sequence comprising IgG; and (ii) a chimeric antigen receptor (CAR) that is designed to be expressed in immune effector cells, such that the macrophages expressing the CAR possess targeted effector activity, and wherein the CAR comprises an extracellular antigen-binding domain, a transmembrane domain and an intracellular domain.
10. A pharmaceutical composition comprising macrophages according to claim 9 and a pharmaceutically acceptable carrier.
11. A method for treating cancer comprising administering to a patient in need thereof a therapeutically effective amount of the pharmaceutical composition according to claim 10 .
12. The method according to claim 8 , wherein the cancer is a primary or a metastatic solid tumor including bladder, brain, breast, cervical, colon, colorectal, glioblastoma, head and neck, kidney, liver, lung, melanoma, ovarian, pancreas, pituitary, prostate, rectal, sarcoma tumors, skin, stomach, testicular, thyroid and uterine cancer.
13. The macrophage according to claim 1 , which is genetically modified to express a peptide which is at least 95% identical to the IL-31 sequence as set forth in SEQ ID No. 1 and which binds to IL-31RA or a fused protein comprising a peptide which is at least 95% identical to the IL-31 sequence as set forth in SEQ ID No. 1 and which binds to IL-31RA.
14. The macrophage according to claim 1 , wherein the peptide which is at least 85% identical to the IL-31 sequence as set forth in SEQ ID No. 1 and which binds to IL-31RA and induces signal transduction through the IL-31RA.
15. The method according to claim 3 , wherein the nucleic acid sequence encodes a peptide which is at least 95% identical to the IL-31 sequence as set forth in SEQ ID No. 1 and which binds to IL-31RA or a fused protein comprising a peptide which is at least 95% identical to the IL-31 sequence as set forth in SEQ ID No. 1 and which binds to IL-31RA.