Exosomal proteins as potential diagnostic markers in advanced non-small cell lung carcinoma.
BACKGROUND
Lung cancer is one of the leading causes of cancer-related death. At the time of diagnosis, more than half of the patients will have disseminated disease and, yet, diagnosing can be challenging. New methods are desired to improve the diagnostic work-up. Exosomes are cell-derived vesicles displaying various proteins on their membrane surfaces. In addition, they are readily available in blood samples where they constitute potential biomarkers of human diseases, such as cancer. Here, we examine the potential of distinguishing non-small cell lung carcinoma (NSCLC) patients from control subjects based on the differential display of exosomal protein markers.


METHODS
Plasma was isolated from 109 NSCLC patients with advanced stage (IIIa-IV) disease and 110 matched control subjects initially suspected of having cancer, but diagnosed to be cancer free. The Extracellular Vesicle Array (EV Array) was used to phenotype exosomes directly from the plasma samples. The array contained 37 antibodies targeting lung cancer-related proteins and was used to capture exosomes, which were visualised with a cocktail of biotin-conjugated CD9, CD63 and CD81 antibodies.


RESULTS
The EV Array analysis was capable of detecting and phenotyping exosomes in all samples from only 10 µL of unpurified plasma. Multivariate analysis using the Random Forests method produced a combined 30-marker model separating the two patient groups with an area under the curve of 0.83, CI: 0.77-0.90. The 30-marker model has a sensitivity of 0.75 and a specificity of 0.76, and it classifies patients with 75.3% accuracy.


CONCLUSIONS
The EV Array technique is a simple, minimal-invasive tool with potential to identify lung cancer patients.
Department of Clinical Biochemistry, Aarhus University Hospital, Aarhus, Denmark.|Department of Biomedicine, Aarhus University, Aarhus, Denmark.
Kristine R
KR
Jakobsen
Department of Clinical Biochemistry, Aarhus University Hospital, Aarhus, Denmark.|Department of Oncology, Aarhus University Hospital, Aarhus, Denmark.
Birgitte S
BS
Paulsen
Department of Clinical Immunology, Aalborg University Hospital, Aalborg, Denmark.
Rikke
R
Bæk
Department of Clinical Immunology, Aalborg University Hospital, Aalborg, Denmark.
Kim
K
Varming
Department of Clinical Biochemistry, Aarhus University Hospital, Aarhus, Denmark.
Boe S
BS
Sorensen
Department of Clinical Immunology, Aalborg University Hospital, Aalborg, Denmark; maljoe@rn.dk.
Malene M
MM
Jørgensen
EV Array; NSCLC; exosomes; extracellular vesicles; lung cancer; phenotyping; plasma; protein microarray