Intestinal expression of programmed death ligand 1

Abstract

Provided herein are methods and compositions for the amelioration of inflammatory disorders comprising the intestinal expression of programmed death ligand 1.

1 .- 13 . (canceled) 
     
     
         14 . An expression vector comprising a PD-L1 nucleic acid, wherein the sequence of said PD-L1 nucleic acid comprises at least one synonymous substitution compared to SEQ ID NO: 2 for detection after administration, and preferably a plurality of synonymous substitutions. 
     
     
         15 . The expression vector according to  claim 14 , wherein said PD-L1 nucleic acid sequence is codon-optimized for expression. 
     
     
         16 . The expression vector according to  claim 14 , wherein said PD-L1 nucleic acid further comprises a nucleic acid sequence that encodes a heterologous sequence, preferably wherein said heterologous sequence comprises an Fc domain, a protein tag, a conjugated therapeutic, or a combination thereof. 
     
     
         17 . (canceled) 
     
     
         18 . The expression vector according to  claim 16 , wherein said Fc domain comprises a human IgG1 Fc region or a portion thereof. 
     
     
         19 . The expression vector according to  claim 18 , wherein said PD-L1 nucleic acid comprises the sequence of SEQ ID NO: 4 or SEQ ID NO: 3. 
     
     
         20 .- 24 . (canceled) 
     
     
         25 . A chitosan derivative nanoparticle comprising an encapsulated expression vector, wherein the expression vector comprises a PD-L1 nucleic acid encoding a human PD-L1 polypeptide for localized intestinal expression of the human PD-L1 polypeptide, wherein the sequence of the PD-L1 nucleic acid comprises a plurality of synonymous substitutions compared to SEQ ID NO 2 for detection after administration. 
     
     
         26 . The chitosan derivative nanoparticle of  claim 25 , comprising chitosan coupled with arginine and a hydrophilic polyol. 
     
     
         27 . The chitosan derivative nanoparticle of  claim 26 , wherein the hydrophilic polyol is glucose. 
     
     
         28 . The chitosan derivative nanoparticle of  claim 25 , wherein the PD-L1 nucleic acid comprises a nucleic acid sequence that is at least about 85% or 95% identical to SEQ ID NO: 3 or about 85% or 95% identical to SEQ ID NO: 4. 
     
     
         29 . The chitosan derivative nanoparticle of  claim 25 , wherein the PD-L1 polypeptide is a soluble human PD-L1 polypeptide comprising or consisting of the IgV domain and the IgC domain of human PD-L1 (amino acids 19-239 of SEQ ID NO: 1). 
     
     
         30 . The chitosan derivative nanoparticle of  claim 25 , wherein the PD-L1 polypeptide is a soluble human PD-L1 polypeptide comprising or consisting of the signal sequence, the IgV domain and the IgC domain of human PD-L1 (amino acids 1-239 of SEQ ID NO: 1). 
     
     
         31 . The chitosan derivative nanoparticle of  claim 25 , wherein the PD-L1 polypeptide is N-terminally fused to a human IgG1 Fc region. 
     
     
         32 . The chitosan derivative nanoparticle of  claim 31 , wherein the PD-L1 polypeptide is fused to the human IgG1 Fc region via an amino acid sequence of (GGGGS)n (SEQ ID NO: 5). 
     
     
         33 . The chitosan derivative nanoparticle of  claim 31 , wherein the human IgG1 Fc is mutated to reduce antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cellular cytotoxicity (CDCC) by altering one or more of the following amino acids in the Fc domain: E233P, L234V, L235A, deletion of G236, A327G, A330S and P331S. 
     
     
         34 . The chitosan derivative nanoparticle of  claim 33 , wherein the PD-L1 nucleic acid comprises the sequence of SEQ ID NO: 4. 
     
     
         35 . The chitosan derivative nanoparticle of  claim 25 , wherein the PD-L1 polypeptide is a membrane-bound PD-L1 polypeptide comprising or consisting of the signal sequence, IgV domain, IgC domain and the transmembrane domain of human PD-L1 (amino acids 1-259 of SEQ ID NO: 1). 
     
     
         36 . The chitosan derivative nanoparticle of  claim 35 , wherein the PD-L1 polypeptide further comprises the cytoplasmic domain of human PD-L1. 
     
     
         37 . The chitosan derivative nanoparticle of  claim 35 , wherein the PD-L1 nucleic acid comprises the sequence of SEQ ID NO: 3. 
     
     
         38 . A method for treating an inflammatory disorder in a patient in need thereof, comprising administering the chitosan derivative nanoparticle of  claim 25  to the gastrointestinal tract of the patient. 
     
     
         39 . The method of  claim 38 , wherein the inflammatory disorder is selected from the group consisting of inflammatory bowel disease, ulcerative colitis, and Crohns disease.