mRNA combination therapy for the treatment of cancer
The present disclosure relates to the use of nucleic acid (e.g., mRNA) combination therapies for the treatment of cancer. The disclosure provides compositions, and methods for their preparation, manufacture, and therapeutic use, wherein those compositions comprise at least two polynucleotides (e.g., mRNAs) in combination wherein the at least two polynucleotides are selected from the group consisting of (i) a polynucleotide encoding an immune response primer (e.g., IL23), (ii) a polynucleotide encoding an immune response co-stimulatory signal (e.g., OX40L), (iii) a polynucleotide encoding a checkpoint inhibitor (e.g., an anti CTLA-4 antibody), and, (iv) a combination thereof. The therapeutic methods disclosed herein comprise, e.g., the administration of a combination therapy disclosed herein for the treatment of cancer, e.g., by reducing the size of a tumor or inhibiting the growth of a tumor, in a subject in need thereof. In some aspects, the combination therapies disclosed herein disclosed are administered intratumorally.
1. A method for increasing immune cell activation in a subject, comprising administering a lipid nanoparticle (LNP) comprising one or more messenger RNAs (mRNAs), wherein the one or more mRNAs are selected from (i) a first mRNA comprising an open reading frame (ORF) encoding a human IL-15 polypeptide and a second mRNA comprising an ORF encoding a human IL-15 receptor alpha (IL-15Ra) polypeptide, and (ii) an mRNA comprising an ORF encoding a fusion protein comprising a human IL-15 polypeptide operably linked to a human IL-15Ra polypeptide, thereby increasing immune cell activation in the subject.
2. The method of claim 1 , wherein the one or more mRNAs comprise the mRNAs of (i).
3. The method of claim 1 , wherein the one or more mRNAs comprise the mRNA of (ii).
4. The method of claim 1 , wherein the IL-15 polypeptide comprises the sequence set forth in SEQ ID NO: 810.
5. The method of claim 1 , wherein the IL-15Ra polypeptide comprises a transmembrane domain.
6. The method of claim 5 , wherein the IL-15Ra polypeptide comprises the sequence set forth in SEQ ID NO: 808.
7. The method of claim 1 , wherein at least one of the one or more mRNAs comprise a miR binding site.
8. The method of claim 1 , wherein the one or more mRNAs comprise uridine nucleosides and the uridine nucleosides in one or more of the mRNAs are chemically modified.
9. The method of claim 8 , wherein the chemically-modified nucleosides are N1-methylpseudouridines (m1ψ).
10. The method of claim 1 , wherein the LNP comprises an ionizable amino lipid, a phospholipid, a sterol, and a PEG-modified lipid.
11. The method of claim 10 , wherein the LNP comprises a molar ratio of (a) 40-60% ionizable amino lipid, 8-16% phospholipid, 30-45% sterol, and 1-5% PEG modified lipid, or (b) 45-65% ionizable amino lipid, 5-10% phospholipid, 25-40% sterol, and 0.5-5% PEG modified lipid.
12. The method of claim 11 , wherein the LNP comprises a molar ratio of 40-60% ionizable amino lipid, 8-16% phospholipid, 30-45% sterol, and 1-5% PEG modified lipid.
13. The method of claim 12 , wherein the LNP comprises a molar ratio of 40-60% Compound 25, 8-16% DSPC, 30-45% cholesterol, and 1-5% PEG DMG.
14. The method of claim 11 , wherein the LNP comprises a molar ratio of 45-65% ionizable amino lipid, 5-10% phospholipid, 25-40% sterol, and 0.5-5% PEG modified lipid.
15. The method of claim 14 , wherein the LNP comprises a molar ratio of 45-65% Compound 25, 5-10% DSPC, 25-40% cholesterol, and 0.5-5% PEG DMG.
16. A method for increasing immune cell activation in a subject, comprising administering two or more LNPs, wherein the two or more LNPs are selected from (i) a first LNP comprising an mRNA comprising an ORF encoding a human IL-15 polypeptide; (ii) a second LNP comprising an mRNA comprising an ORF encoding a human IL-15Ra polypeptide; and (iii) both the first and second LNPs, thereby increasing immune cell activation in the subject.
17. The method of claim 16 , wherein the two or more LNPs comprise the LNP of (i).
18. The method of claim 16 , wherein the two or more LNPs comprise the LNP of (ii).
19. The method of claim 16 , wherein the two or more LNPs comprise the LNPs of (iii).
20. The method of claim 16 , wherein the IL-15 polypeptide comprises the sequence set forth in SEQ ID NO: 810.
21. The method of claim 16 , wherein the IL-15Ra polypeptide comprises a transmembrane domain.
22. The method of claim 21 , wherein the IL-15Ra polypeptide comprises the sequence set forth in SEQ ID NO: 808.
23. The method of claim 16 , wherein the mRNAs comprise a miR binding site.
24. The method of claim 16 , wherein the mRNAs comprise uridine nucleosides and the uridine nucleosides in the mRNAs are chemically modified.
25. The method of claim 16 , wherein the chemically-modified nucleosides are N1-methylpseudouridines (m1ψ).
26. The method of claim 16 , wherein the LNP comprises an ionizable amino lipid, a phospholipid, a sterol, and a PEG-modified lipid.
27. The method of claim 26 , wherein the LNP comprises a molar ratio of (a) 40-60% ionizable amino lipid, 8-16% phospholipid, 30-45% sterol, and 1-5% PEG modified lipid, or (b) 45-65% ionizable amino lipid, 5-10% phospholipid, 25-40% sterol, and 0.5-5% PEG modified lipid.
28. The method of claim 27 , wherein the LNP comprises a molar ratio of 40-60% ionizable amino lipid, 8-16% phospholipid, 30-45% sterol, and 1-5% PEG modified lipid.
29. The method of claim 28 , wherein the LNP comprises a molar ratio of 40-60% Compound 25, 8-16% DSPC, 30-45% cholesterol, and 1-5% PEG DMG.
30. The method of claim 27 , wherein the LNP comprises a molar ratio of 45-65% ionizable amino lipid, 5-10% phospholipid, 25-40% sterol, and 0.5-5% PEG modified lipid.
31. The method of claim 30 , wherein the LNP comprises a molar ratio of 45-65% Compound 25, 5-10% DSPC, 25-40% cholesterol, and 0.5-5% PEG DMG.