Method for monitoring cancer and/or inflammatory reaction based on RelB phosphorylation
View Patent ↗The present Inventors demonstrated that the RelB subunit of NFκB plays a crucial role in promoting cell migration. More precisely, they identified that this pro-migratory activity is mediated by the activation of the NFκB pathway through RelB phosphorylation at serine 472. In a first aspect, the present invention proposes to monitor the activation of the NFκB pathway by following the phosphorylation status of said serine. Also, the present invention discloses methods and kits for prognosing the evolution of a disease involving cell migration in a subject—treated or not—suffering thereof, based on the detection of said RelB-S472 phosphorylation.
1. An in vitro method, comprising:
a) providing a sample; and
b) detecting RelB (SEQ ID NO: 1) phosphorylated on serine 472 or a RelB-homologue having at least 80% identity to (SEQ ID NO: 1) and phosphorylated on a corresponding serine, in said sample.
2. The method of claim 1 , wherein the phosphorylated RelB (SEQ ID NO: 1) or the phosphorylated RelB-homologue having at least 80% identity to SEQ ID NO: 1 is detected by contacting the sample with a reagent that binds specifically to the phosphorylated RelB (SEQ ID NO: 1) or to the phosphorylated RelB-homologue, and detecting binding of the reagent to the phosphorylated RelB (SEQ ID NO: 1) or to the phosphorylated RelB-homologue having at least 80% identity to SEQ ID NO: 1.
3. The method of claim 2 , wherein the reagent is selected from the group consisting of an antibody, an intrabody, and an aptamer.
4. The method of claim 3 , wherein the antibody is a monoclonal antibody.
5. The method of claim 1 , wherein RelB (SEQ ID NO: 1) phosphorylated on serine 472 is detected.
6. The method of claim 4 , wherein RelB (SEQ ID NO: 1) phosphorylated on serine 472 is detected.
7. The method of claim 1 , wherein the phosphorylated RelB (SEQ ID NO: 1) or the phosphorylated RelB-homologue having at least 80% identity to SEQ ID NO: 1 is detected by mass spectrometry using a heavy-isotope labeled peptide.
8. The method of claim 1 , further comprising:
c) measuring the phosphorylation level of serine 472 of the RelB protein (SEQ ID NO:1), or of the corresponding serine of the RelB-homolog, in the sample.
9. A method for treating cancer, comprising:
a) providing a biological sample from a subject,
b) detecting the presence of phosphorylated RelB (SEQ ID NO: 1) or of a phosphorylated RelB-homologue having at least 80% identity to SEQ ID NO: 1 in the sample by the method of claim 1 , and
c) administering to the subject an efficient dose of a treatment.
10. The method of claim 9 , wherein the cancer is selected from the group consisting of breast cancer, glioblastoma, and prostate cancer.
11. An in vitro screening method comprising:
a) providing a candidate molecule,
b) incubating the candidate molecule with RelB (SEQ ID NO: 1) in the presence of a IkB kinase in phosphorylating conditions, and
c) detecting whether phosphorylation of serine 472 of RelB (SEQ ID NO: 1) occurs.