Protein Microarray Signature of Autoantibody Biomarkers for the Early Detection of Breast Cancer

Abstract

Cancer patients spontaneously generate autoantibodies (AAb) to tumor-derived proteins.. To detect AAb, we have probed novel high-density custom protein microarrays (NAPPA) expressing 4,988 candidate tumor antigens with sera from patients with early stage breast cancer (IBC), and bound IgG was measured. We used a three-phase serial screening approach. First, a pre-screen was performed to eliminate uninformative antigens. Sera from stage I–III IBC (n=53) and healthy women (n=53) were screened for AAb to all 4,988 protein antigens. Antigens were selected if the 95th percentile of signal of cases and controls were significantly different (p<0.05) and if the number of cases with signals above the 95th percentile of controls was significant (p<0.05). These 761 antigens were screened using an independent set of IBC sera (n=51) and sera from women with benign breast disease (BBD) (n=39). From these, 119 antigens had a partial area under the ROC curve (p<0.05), with sensitivities ranging from 9–40% at >91% specificity. 28 of these antigens were confirmed using an independent serum cohort (n=51 cases/38 controls, p<0.05). Using all 28 AAb, a classifier was identified with a sensitivity of 80.8% and a specificity of 61.6% (AUC=0.756). These are potential biomarkers for the early detection of breast cancer.

A Multiparametric Serum Marker Panel as a Complementary Test to Mammography for the Diagnosis of Node-Negative Early-Stage Breast Cancer and DCIS in Young Women

“…They are biochemically well-characterized, and many reagents and techniques are available for their detection, simplifying assay development. Until very recently, several specific autoantibodies have been detected in the sera of patients with advanced-stage breast cancer (14,(39)(40)(41)(42)(43)(44)(45)(46)(47)(48)(49)(50)(51)(52)(53)(54)(55)(56)(57)(58). However, very few studies have attempted to identify and/or evaluate autoantibodies in node-negative earlystage PBC ( 2 cm) or, more importantly, in preinvasive breast cancer, for which mammography's sensitivity is decreased.…”

Overexpressed oncogenic tumor-self antigens

“…Early clinical support for the promise of TPD52 as a vaccine target antigen came from the identification of circulating serum antibodies with specificity for TPD52 in patients with breast cancer. 74,75 Expression of TPD52 in cancer TPD52 is an overexpressed tumor self-protein actively involved in transformation, leading to increased proliferation and metastasis. TPD52 overexpression has been demonstrated in several human malignancies including breast, [76][77][78] prostate, [79][80][81] and ovarian carcinomas.…”

Emerging technology of in situ cell free expression protein microarrays

“…Protein microarrays are a great tool for the large-scale analysis of both functional genomics and proteomics (Predki, 2004;Lueking et al, 2005). Currently protein arrays are extensively used for various applications such as antibody profiling, biomarker identification, protein-protein interactions, and enzymatic assays (Kawahashi et al, 2003;Feilner et al, 2005;Ramachandran et al, 2008b;Anderson et al, 2011;Hu et al, 2011;Ramani et al, 2012).…”