Cross-Species Antibody Microarray Interrogation Identifies a 3-Protein Panel of Plasma Biomarkers for Early Diagnosis of Pancreas Cancer

Abstract

Purpose Pancreatic ductal adenocarcinoma (PDA) is the fourth leading cause of cancer death in the United States and its incidence is on the rise. Advanced disease is nearly uniformly lethal, emphasizing the need to identify PDA at its earliest stages. To discover early biomarkers of PDA, we evaluated the circulating proteome in murine preinvasive and invasive plasma samples and human pre-diagnostic and diagnostic samples. Experimental Design Using a customized antibody microarray platform containing >4000 features, we interrogated plasma samples spanning preinvasive and invasive disease from a highly faithful mouse model of PDA. In parallel, we mined pre-diagnostic plasma from women in the Women’s Health Initiative (WHI) who would later succumb to PDA together with matched, cancer-free control samples. Samples collected after an establishing diagnosis of PDA were also interrogated to further validate markers. Results We identified ERBB2 and TNC in our cross-species analyses and multiple antibodies identified ESR1 in pre-diagnostic plasma from people that succumb to PDA. This 3-marker panel had an AUC of 0.86 (0.76–0.96, 95% confidence interval (CI)) for the diagnostic cohort that increased to 0.97 (0.92–1.0, 95% CI) with CA19-9 included. The 3-marker panel also had an AUC of 0.68 (0.58–0.77, 95% CI) for the pre-diagnostic cohort. Conclusions We identified potential disease detection markers in plasma up to 4 years prior to death from PDA with superior performance to CA19-9. These markers might be especially useful in high-risk cohorts to diagnose early, resectable disease, particularly in patients that do not produce CA19-9.

Sequential Validation of Blood-Based Protein Biomarker Candidates for Early-Stage Pancreatic Cancer

“…Although promising PDAC plasma protein biomarkers have been previously reported (11,12,14), this is the first study where a proteomics-based discovery, performed using both human prediagnostic and mouse early-stage PDAC plasma samples, was followed by sequential validation of the identified biomarker candidates in multiple independent sets of samples from resectable PDAC patients and matched controls. By applying a rigorous statistical method, we were able to build a model with statistically significantly improved performance than CA19-9 alone, which was blindly tested in an independent sample set.…”

“…As a result, there is substantial ongoing effort to identify additional serological biomarkers that complement CA19-9 for the detection of early-stage PDAC. Markers investigated include proteins (9)(10)(11)(12)(13)(14), metabolites (15)(16)(17), autoantibodies (18,19), miRNAs (20)(21)(22), markers with aberrant glycosylation (CA19-9 and Tn antigens) (6,23,24), and exosomes (25). In view of the wide array of potential biomarkers, there is a need for systematic evaluation of candidates using CA19-9 as an anchor marker given its established performance.…”

Genetics and biology of pancreatic ductal adenocarcinoma

“…Moreover, genetic alterations spanning the whole tumor genome have been identified in these serum-derived exosomes , underscoring the potential translational significance of exosome-based detection methods combined with genetic profiling for accurate detection and profiling of much earlier stage malignancies. Finally, cross-species serum proteomic analysis has proven to be another productive avenue for the identification of novel protein biomarkers that can detect early PDAC neoplasms well before the onset of clinical signs and symptoms of PDAC (Faca et al 2008;Day et al 2015;Mirus et al 2015). Such protein biomarkers may add sensitivity and specificity when combined with ctDNA and exosome strategies and accelerate the development of reliable early detection and liquid biopsy technologies.…”

Overexpression of microRNA-5100 decreases the aggressive phenotype of pancreatic cancer cells by targeting PODXL

“…Pancreatic cancer is the fourth leading cause of cancer mortality in the United States (1,2). Although efforts to reduce risk factors such as smoking, obesity, and high meat consumption and to improve early detection have been made, pancreatic cancer is still formidable because of its aggressive metastatic ability (3)(4)(5)(6).…”