Gitrl fusion proteins and uses thereof

Abstract

The disclosure provides GITRL fusion polypeptide subunits comprising an IgG Fc domain, a trimerization domain, and the receptor binding domain of GITR ligand, where the fusion polypeptide subunits can self-assemble into hexameric proteins. Also provided are methods of making fusion polypeptide subunits and hexameric proteins, and methods of use, e.g., treatment of cancer.

1 - 136 . (canceled) 
     
     
         137 : A method of treating a solid tumor in a subject in need thereof, comprising administering to the subject a hexameric protein comprising six single-chain polypeptide subunits each comprising:
 an IgG Fc domain;   a functional trimerization domain derived from human Coronin 1a; and   a receptor binding domain of a Glucocorticoid-Induced TNF Receptor Ligand (GITRL), wherein the GITRL receptor binding domain comprises the amino acid sequence of SEQ ID NO: 35, and wherein residue 112 of SEQ ID NO: 35 is an aspartyl residue.   
     
     
         138 : The method of  claim 137 , wherein the IgG Fc domain comprises an IgG1, IgG2, IgG3, IgG4, IG4P Fc domain, or variants thereof. 
     
     
         139 : The method of  claim 137 , wherein the IgG Fc domain includes an IgG1 hinge region, an IgG4 hinge region, or variants thereof at its amino terminus. 
     
     
         140 : The method of  claim 139 , wherein the IgG hinge region comprises a mutation that confers complete inter heavy chain disulfide bond formation. 
     
     
         141 : The method of  claim 137 , wherein administration of the hexameric protein inhibits tumor growth, promotes tumor reduction, or both. 
     
     
         142 : The method of  claim 137  further comprising administering to the subject an OX40 agonist to the subject. 
     
     
         143 : The method of  claim 142 , wherein the OX40 agonist is one or more of an OX40 ligand fusion protein and an anti-OX40 antibody. 
     
     
         144 : The method of  claim 143 , wherein the OX40 ligand fusion protein is MEDI6383. 
     
     
         145 : The method of  claim 143 , wherein the anti-OX40 antibody is MEDI0562. 
     
     
         146 : A method of treating cancer in a subject in need thereof, comprising administering to the subject:
 a) an effective amount of a hexameric protein comprising six single-chain polypeptide subunits each comprising:
 an IgG Fc domain, 
 a functional trimerization domain derived from human Coronin 1a, and a receptor binding domain of a Glucocorticoid-Induced TNF Receptor Ligand (GITRL), wherein the GITRL receptor binding domain comprises the amino acid sequence of SEQ ID NO: 35 and wherein residue 112 of SEQ ID NO: 35 is an aspartyl residue; and 
   b) a T-cell priming agent.   
     
     
         147 : The method of  claim 146 , wherein the T-cell priming agent is a combination of a DNA vaccine and an adjuvant. 
     
     
         148 : The method of  claim 147 , wherein the T-cell priming agent is a combination of E7 Synthetic Long Peptide (SLP) and CpG Oligodeoxynucleotide. 
     
     
         149 : The method of  claim 146 , wherein the subject is a human subject or a canine subject. 
     
     
         150 : The method of  claim 146 , wherein the cancer is a solid tumor. 
     
     
         151 : The method of  claim 146 , wherein the subject has colorectal cancer, breast cancer, hepatocellular cancer, non-small cell lung cancer (NSCLC), small cell lung cancer, mesothelioma, head and neck cancer, gastric cancer, pancreatic cancer, melanoma, uveal melanoma, renal cancer, ovarian cancer, cervical cancer, glioblastoma, testicular cancer, thyroid cancer, prostate cancer, oesophageal cancer, bladder cancer, squamous cell carcinoma of the head and neck (SCCHN), colorectal cancer (CRC), haematological cancer, or a combination thereof. 
     
     
         152 : The method of  claim 151 , wherein the cancer is a haematological cancer selected from the group consisting of Hodgkins lymphoma, non-Hodgkins lymphoma, multiple myeloma, acute lymphoblastic leukemia, acute myelogenous leukemia, chronic lymphocytic leukemia, and chronic myelogenous leukemia. 
     
     
         153 : A polynucleotide comprising a nucleic acid that encodes a polypeptide subunit comprising: an IgG Fc domain; a functional trimerization domain derived from human Coronin 1a; and a receptor binding domain of a Glucocorticoid-Induced TNF Receptor Ligand (GITRL), wherein the GITRL receptor binding domain comprises the amino acid sequence of SEQ ID NO: 35, and wherein residue 112 of SEQ ID NO: 35 is an aspartyl residue. 
     
     
         154 : The polynucleotide of  claim 153  comprising SEQ ID NO: 5. 
     
     
         155 : A vector comprising the polynucleotide of  claim 153 . 
     
     
         156 : A host cell comprising the polynucleotide of  claim 154 .