Tumor-specific ifna secretion by car t-cells to reprogram the solid tumor microenvironment

Abstract

The present application relates to fusion proteins, chimeric antigen bearing cells expressing fusion proteins and compositions comprising chimeric antigen bearing cells expressing fusion proteins. The application further relates to methods of using the fusion proteins, cells and compositions for modulating an immune response.

1 .- 10 . (canceled) 
     
     
         11 . A chimeric polypeptide comprising:
 an extracellular domain of a PD-1 protein;   a spacer; and   an interferon-α2 protein.   
     
     
         12 . The chimeric polypeptide of  claim 11 , wherein the PD-1 protein comprises a PD-1 extracellular domain of a protein encoded by a nucleotide sequence having at least 95% sequence identity with any one of SEQ ID NOs: 14, 16 or 17. 
     
     
         13 . The chimeric polypeptide of  claim 12 , wherein the PD-1 protein comprises a PD-1 extracellular domain of a protein encoded by the nucleotide sequence of SEQ ID NO:14. 
     
     
         14 . The chimeric polypeptide of  claim 12 , wherein the PD-1 protein comprises a PD-1 extracellular domain of a protein encoded by the nucleotide sequence of SEQ ID NO:16. 
     
     
         15 . The chimeric polypeptide of  claim 12 , wherein the PD-1 protein comprises a PD-1 extracellular domain of a protein encoded by the nucleotide sequence of SEQ ID NO:17. 
     
     
         16 . The chimeric polypeptide of  claim 11 , wherein the extracellular domain of a mutant PD-1 protein comprises an amino acid sequence having at least 95% sequence identity with of SEQ ID NO:05, or SEQ ID NO:06. 
     
     
         17 . The chimeric polypeptide of  claim 16 , wherein the extracellular domain of a mutant PD-1 protein comprises the amino acid sequence of SEQ ID NO:05. 
     
     
         18 . The chimeric polypeptide of  claim 16 , wherein the extracellular domain of a mutant PD-1 protein comprises the amino acid sequence of SEQ ID NO:06. 
     
     
         19 . The chimeric polypeptide of  claim 11 , wherein the interferon-α2 protein is IFNα2a. 
     
     
         20 . The chimeric polypeptide of  claim 19 , wherein the IFNα2a is encoded by the nucleotide sequence of SEQ ID NO:01. 
     
     
         21 . The chimeric polypeptide of  claim 11 , wherein the spacer comprises at least 3 consecutive glycine residues. 
     
     
         22 . The nucleic acid of  21 , wherein the spacer comprises the amino acid sequence of SEQ ID NO:07. 
     
     
         23 . The nucleic acid of  21 , wherein the spacer comprises the amino acid sequence of SEQ ID NO:08. 
     
     
         24 . The nucleic acid of  21 , wherein the spacer comprises the amino acid sequence of SEQ ID NO:09. 
     
     
         25 . A method of preparing a cell comprising a chimeric polypeptide, comprising: introducing into the cell a nucleic acid encoding the polypeptide of  claim 11 . 
     
     
         26 . The method of  claim 25 , wherein the cell is selected from a lymphocyte, a NK cell, a precursor T-cell, a stem cell, a hematopoietic stem cell, a NK cell, a neuronal stem cell, or a B cell. 
     
     
         27 . The method of  claim 25 , wherein the cell is a CD8+ T cell or a CD4+ T cell. 
     
     
         28 . The method of  claim 25 , wherein the cell comprises an chimeric antigen receptor (CAR). 
     
     
         29 . The method of  claim 28 , wherein the CAR comprises an anti-CD19 CAR.