Abstract 1080: Nanotechnology-enhanced proteomics analysis of a unique prospective collection of blood samples for the discovery and validation of early detection biomarkers for Non-Small Cell Lung Cancer (NSCLC).
Authors: Fredolini Claudia , Luchini Alessandra , Paris Luisa , Magni Ruben , Petricoin Emanuel F. , Darakjy Salima , Gaziano J. Michael , Sesso Howard D. , Liotta Lance A. , Perera Frederica P. 
Published: August 2014

Abstract
BACKGROUND. Early identification of individuals at high risk to develop lung cancer would be a crucial requirement for a successful prevention strategy. The novelty of this study lies in our ability to discover and validate biomarkers for subclinical disease in blood samples obtained from healthy individuals up to 25 years prior to lung cancer diagnosis. Plasma samples were collected from healthy individuals who later developed Non-Small Cell Lung Cancer (NSCLC) (cases) matched with individuals who did not develop cancer after the same period of follow up (controls). We studied the influence of smoking status (non-smokers, smokers and former smokers) on the pattern of blood borne biomarkers in order to test the hypothesis that the developing tumor might have distinct molecular origins in the three classes.
METHODS. Study subjects were drawn from the longitudinal Physicians’ Health Study (PHS). We analyzed plasma samples from 42 cases and 42 controls (16 to 284 month follow up). Small proteins and peptides were purified from plasma using N-isopropylacrylamide (NIPAm) core shell hydrogel particles functionalized with Cibacron blue F3GA and vinylsulfonic acid as chemical baits. Proteins eluted from the particles were analyzed using LTQ-Orbitrap mass spectrometer (Thermo Fisher).
RESULTS. 67, 83 and 75 candidate biomarkers of early NSCLC were identified for non-smokers, former smokers and smokers, respectively. An optimal combination of predictive biomarkers was selected for each group. Receiver-operating characteristic (ROC) curves were utilized to evaluate the predictive value of selected biomarkers. In non-smokers, a panel of 8 proteins including Keratin 6B and endostatin had 82% sensitivity and 91% specificity (AUC 0.926) in predicting an individual developing cancer. In former smokers, a combination of 8 proteins (including malic enzyme 2 and mitochondrial malate dehydrogenase) had 88% sensitivity and 82% specificity (AUC 0.901). In smokers, 10 selected biomarkers (including fructose-bisphosphate aldolase A and glyceraldehyde-3-phosphate dehydrogenase) had 75% sensitivity and 75% specificity (AUC 0.833). Differential plasma protein profiles of smokers or former smokers that developed cancer included a high number of proteins involved in the response to high oxidative stress. Oxidative stress, together with inflammation, could be the driving pre requisite for lung cancer development in individuals exposed to smoke. Interestingly, endostatin (a known inhibitor of angiogenesis) was over-represented (Chi-Square p &lt; 0.031) in cases versus controls independent of smoking status. The predictive value of identified candidate biomarkers will be verified in a larger cohort of independent blinded samples from the same prospective collection study.
Citation Format: Claudia Fredolini, Alessandra Luchini, Luisa Paris, Ruben Magni, Emanuel F. Petricoin, Salima Darakjy, J. Michael Gaziano, Howard D. Sesso, Lance A. Liotta, Frederica P. Perera. Nanotechnology-enhanced proteomics analysis of a unique prospective collection of blood samples for the discovery and validation of early detection biomarkers for Non-Small Cell Lung Cancer (NSCLC). [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1080. doi:10.1158/1538-7445.AM2013-1080