Combinations of mrnas encoding immune modulating polypeptides and uses thereof

Abstract

The disclosure relates to compositions and methods for the preparation, manufacture and therapeutic use of combinations of immunomodulatory polynucleotides (e.g., mRNAs) encoding an immune response primer polypeptide (e.g., an interleukin 23 (IL-23) polypeptide or an interleukin 36γ (IL-36-gamma) polypeptide), and an immune response co-stimulatory signal polypeptide (e.g., an OX40L polypeptide).

What is claimed is: 
     
         1 . A method for treating cancer in a subject, comprising administering intratumorally to the subject a lipid nanoparticle (LNP) comprising: (i) a first mRNA comprising an open reading frame (ORF) encoding a human IL-23 polypeptide and (ii) a second mRNA comprising an ORF encoding a human OX40L polypeptide. 
     
     
         2 . The method of  claim 1 , wherein the IL-23 polypeptide is a fusion protein comprising an IL-12p40 polypeptide operably linked, with or without a linker, to an IL-23p19 polypeptide. 
     
     
         3 . The method of  claim 2 , wherein the IL-12p40 polypeptide is operably linked with a linker to the IL-23p19 polypeptide. 
     
     
         4 . The method of  claim 3 , wherein the linker is a Gly/Ser linker. 
     
     
         5 . The method of  claim 2 , wherein the fusion protein comprises a signal peptide. 
     
     
         6 . The method of  claim 5 , wherein the signal peptide is a human IL-12p40 signal peptide or a human IL-23p19 signal peptide. 
     
     
         7 . The method of  claim 3 , wherein the IL-23 polypeptide comprises the amino acid sequence as set forth in SEQ ID NO: 140. 
     
     
         8 . The method of  claim 1 , wherein the OX40L polypeptide comprises the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 21 
     
     
         9 . The method of  claim 1 , wherein the first mRNA encodes the amino acid sequence as set forth in SEQ ID NO: 140; and (ii) the second mRNA encodes the amino acid sequence as set forth in SEQ ID NO: 21. 
     
     
         10 . The method of  claim 1 , wherein each of the first and second mRNAs comprise a 5′cap, a 5′untranslated region (UTR), a 3′UTR comprising a miR-122 binding site and a polyA tail. 
     
     
         11 . The method of  claim 1 , wherein each of the first and second mRNAs are fully modified with chemically-modified uridines. 
     
     
         12 . The method of  claim 1 , wherein the LNP comprises an ionizable amino lipid, a phospholipid, a sterol, and a PEG-modified lipid. 
     
     
         13 . The method of  claim 12 , wherein the LNP comprises a molar ratio of about 20-60% ionizable amino lipid, about 5-25% phospholipid, about 25-55% sterol, and about 0.5-1.5% PEG-modified lipid. 
     
     
         14 . The method of  claim 13  wherein the lipid nanoparticle comprises (i) a molar ratio of 40-60% ionizable amino lipid, 8-16% phospholipid, 30-45% sterol, and 1-5% PEG modified lipid; or (ii) 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 12 , wherein the ionizable lipid comprises a compound having the formula: 
       
         
           
           
               
               
           
         
       
     
     
         16 . The method of  claim 12 , wherein the PEG lipid comprises PEG-DMG, the phospholipid comprises DPSC, and the structural lipid comprises cholesterol. 
     
     
         17 . The method of  claim 1 , wherein the LNP comprises a third mRNA comprising an ORF encoding a polypeptide of interest. 
     
     
         18 . The method of  claim 1 , comprising administering a checkpoint inhibitor polypeptide, wherein the checkpoint inhibitor polypeptide inhibits PD1, PD-L1, CTLA4, or a combination thereof, and wherein the checkpoint inhibitor polypeptide is an antibody or antigen-binding fragment thereof. 
     
     
         19 . The method of  claim 18 , wherein the antibody is an anti-CTLA4 antibody or antigen-binding fragment thereof that specifically binds CTLA4, an anti-PD1 antibody or antigen-binding fragment thereof that specifically binds PD1, an anti-PD-L1 antibody or antigen-binding fragment thereof that specifically binds PD-L1, or a combination thereof. 
     
     
         20 . The method of  claim 19 , wherein the anti-PD-L1 antibody is atezolizumab, avelumab, or durvalumab, wherein the anti-CTLA-4 antibody is tremelimumab or ipilimumab, and wherein the anti-PD1 antibody is nivolumab or pembrolizumab.