Tumor necrosis factor 1B (TNF-1B) mutants and methods of use thereof to screen for candidate therapeutic compounds
Disclosed are tumor necrosis factor receptor 1B (TNFR-1B) signaling targets and TNFR-1B mutants and their uses for treatment of diseases and disorders.
1. A mutant of tumor necrosis factor receptor 1B (TNFR-1B) (SEQ ID NO:3) that modulates the binding of TNFR-1B to one or more of tumor necrosis factor receptor alpha (TNFα), inducible costimulatory ligand (ICOS-L), and mucosal vascular addressing cell adhesion molecule 1 (MadCAM-1), wherein the mutant is selected from the group consisting of mutants K42D, T48A, N171D, S79D, R113D, L114A, R119D, K120D, D58A, R19D, S59D, L64D, R77A, S107D, R119A, K120A, R129A, V138D, K140A, 1156D, 1168D, N171A and M174D, wherein mutant numbering is based on the mature form of TNFR-1B in which the signal peptide at residues 1-22 has been removed.
2. The mutant of claim 1 , selected from the group consisting of mutants K42D, T48A and N171D that, compared to TNFR-1B, have reduced binding to TNFα, ICOS-L and MadCAM-1.
3. The mutant of claim 1 , selected from the group consisting of mutants S79D, R113D, L114A, R119D and K120D that, compared to TNFR-1B, have reduce binding to ICOS-L and MadCAM-1, but not to TNFα.
4. The mutant of claim 1 , which is mutant D58A that, compared to TNFR-1B, has reduced binding to TNFα and ICOS-L, but not to MadCAM-1.
5. The mutant of claim 1 , selected from the group consisting of mutants R19D, S59D, L64D, R77A, S107D, R119A, K120A, R129A, V138D, K140A, 1156D, 1168D, N171A and M174D that, compared to TNFR-1B, predominately reduce binding to MadCAM-1.
6. A fusion protein comprising the mutant of claim 1 and an immunoglobulin Fc sequence.
7. The fusion protein of claim 6 , wherein the immunoglobulin is an IgG.
8. The fusion protein of claim 6 , wherein the immunoglobulin is IgG1.
9. A pharmaceutical composition comprising the fusion protein of claim 6 and a pharmaceutically acceptable carrier.
10. A method for screening for a candidate compound for treating a disease or disorder selected from the group consisting of rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, plaque psoriasis, juvenile idiopathic arthritis, inflammation, autoimmune disease, immune disorder, Crohn's disease, ulcerative colitis and inflammatory bowel disease, the method comprising testing the compound to determine if the compound modulates the interaction between one or more of TNFR-1B and ICOS-L, TNFR-1B and MadCAM-1, and TNFR-1B and ISLR2, wherein a compound that is tested and determined to modulate the interaction between one or more of TNFR-1B and ICOS-L, TNFR-1B and MadCAM-1, or TNFR-1B and ISLR2 is a candidate compound for treating a disease or disorder selected from the group consisting of rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, plaque psoriasis, juvenile idiopathic arthritis, inflammation, autoimmune disease, immune disorder, Crohn's disease, ulcerative colitis and inflammatory bowel disease.
11. The method of claim 10 , wherein binding between TNFR-1B and ICOS-L, TNFR-1B and MadCAM-1, and/or TNFR-1B and ISLR2 is determined in the presence of the candidate compound and in the absence of the candidate compound, and wherein a change in the binding between TNFR-1B and ICOS-L, TNFR-1B and MadCAM-1, and/or TNFR-1B and ISLR2 in the presence of the candidate compound indicates that the candidate compound modulates the interaction between TNFR-1B and ICOS-L, TNFR-1B and MadCAM-1, and/or TNFR-1B and ISLR2.
12. The method of claim 10 , wherein the compound is a non-naturally occurring small molecule of 2,000 daltons or less, or an antibody or an antibody fragment.