1. Department of Pathology, University of Pittsburgh; UPMC Hillman Cancer Center, Pittsburgh, PA.
2. Department of Immunology, University of Pittsburgh; UPMC Hillman Cancer Center, Pittsburgh, PA.
3. Department of Biostatistics, University of Pittsburgh; UPMC Hillman Cancer Center, Pittsburgh, PA.
4. Department of Medicine, University of Pittsburgh; UPMC Hillman Cancer Center, Pittsburgh, PA.
5. Department of Pharmacology and Chemical Biology, University of Pittsburgh; UPMC Hillman Cancer Center, Pittsburgh, PA.
6. VAPHS, Pittsburgh, PA.
7. BioZyme Inc, Apex, NC.
8. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA.
9. Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA.
Background: Increases in expression of ADAM10 and ADAM17 genes and proteins are inconsistently found in cancer lesions, and are not validated as clinically useful biomarkers. The enzyme-specific proteolytic activities, which are solely mediated by the active mature enzymes, directly reflect enzyme cellular functions and might be superior biomarkers than the enzyme gene or protein expressions, which comprise the inactive proenzymes and active and inactivated mature enzymes.
Methods: Using a recent modification of the proteolytic activity matrix analysis (PrAMA) measuring specific enzyme activities in cell and tissue lysates, we examined the specific sheddase activities of ADAM10 (ADAM10sa) and ADAM17 (ADAM17sa) in human non-small cell lung-carcinoma (NSCLC) cell lines, patient primary tumors and blood exosomes, and the noncancerous counterparts.
Results: NSCLC cell lines and patient tumors and exosomes consistently showed significant increases of ADAM10sa relative to their normal, inflammatory and/or benign-tumor controls. Additionally, stage IA-IIB NSCLC primary tumors of patients who died of the disease exhibited greater increases of ADAM10sa than those of patients who survived 5 years following diagnosis and surgery. In contrast, NSCLC cell lines and patient tumors and exosomes did not display increases of ADAM17sa.
Conclusions: This study is the first to investigate enzyme-specific proteolytic activities as potential cancer biomarkers. It provides a proof-of-concept that ADAM10sa could be a biomarker for NSCLC early detection and outcome prediction. To ascertain that ADAM10sa is a useful cancer biomarker, further robust clinical validation studies are needed.
Keywords: Lung cancer, Tumor tissue, Blood exosomes, Lysate, Fluorogenic peptide substrate, Proteolytic activity matrix analysis, ADAM10, ADAM17, Sheddase activity, Cancer biomarker