Application of N-terminomics to NETosis in inflammation
Provided herein are protein biomarkers for NETosis related diseases, including proteases. Also provided are substrate sequences of such proteases and uses thereof.
1. A method for identifying a subject having a NETosis-related inflammatory condition, wherein NETosis is Neutrophil cell death forming Extracellular Traps, comprising:
(a) determining information on NET-associated protease content from a sample from the subject, wherein the protease is selected from the group consisting of neutrophil elastase (NE), cathepsin G (CG), proteinase 3 (PR3), and neutrophil secreted protein 4 (NSP4);
(b) comparing the level of NET-associated protease content with that of a comparable sample from a healthy subject;
(c) identifying the subject as having a NETosis-related inflammatory condition when the level of the NET-associated protease content of the sample is greater than that of the comparable sample from the healthy subject; and
(d) treating the NETosis-related inflammatory condition comprising administering to the subject a protease inhibitor, wherein the protease inhibitor inhibits cleavage of peptide substrate comprising a sequence selected from the group consisting of SEQ ID NOs. 1-180.
2. The method of claim 1 , wherein the sample is a blood sample.
3. The method of claim 1 , wherein the inflammatory condition is selected from the group consisting of infection, systemic lupus erythematosus, rheumatoid arthritis, cystic fibrosis, deep vein thrombosis, pre-eclampsia, periodontitis, appendicitis, tuberculosis, and Crohn's disease.
4. The method of claim 1 , wherein treating comprises further administering to the subject a steroid or non-steroidal anti-inflammatory drug.
5. The method of claim 1 , wherein treating comprises further administering to the subject an antibiotic.
6. The method of claim 1 , wherein obtaining information comprises obtaining a sample from the subject.
7. The method of claim 1 , wherein obtaining information comprises performing protease content measurement on the sample.
8. The method of claim 7 , wherein the protease content assessment is determined by enzyme-linked immunosorbent assay (ELISA), mass spectrometry, chromatography, electrophoresis, radioimmunoas say, flow cytometry, fluorescence activated cell sorting (FACS), or western blotting.
9. The method of claim 8 , wherein the protease content difference between the sample and the comparable sample is +10%, +20%, +25%, +30%, +40%, +50%, +75% or +100%.
10. The method of claim 1 , wherein the subject is a human.
11. The method of claim 1 , wherein the subject is a non-human mammal.