Prediction of colorectal cancer diagnosis based on circulating plasma proteins

Abstract

Non-invasive detection of colorectal cancer with blood-based markers is a critical clinical need. Here we describe a phased mass spectrometry-based approach for the discovery, screening, and validation of circulating protein biomarkers with diagnostic value. Initially, we profiled human primary tumor tissue epithelia and characterized about 300 secreted and cell surface candidate glycoproteins. These candidates were then screened in patient systemic circulation to identify detectable candidates in blood plasma. An 88-plex targeting method was established to systematically monitor these proteins in two large and independent cohorts of plasma samples, which generated quantitative clinical datasets at an unprecedented scale. The data were deployed to develop and evaluate a five-protein biomarker signature for colorectal cancer detection.

Identification and Validation of Stage‐Associated Serum Biomarkers in Colorectal Cancer Using MS‐Based Procedures

“…For instance, IGF2 and ECM1 were significantly increased in the CRC sera and could be considered as "back-up" proteins for the development of new predictive signatures and for the identification of distinct subpopulations of colorectal cancer. [32,33] Colorectal cancer is a heterogeneous disease consisting of at least four to five different subtypes with quite different molecular features. [31] In agreement with our results, ECM1 seems to be overexpressed preferentially at late stages ( Figure 4).…”

“…[33] For the implementation of a diagnostic panel, final testing should include a prospective screening cohort. To get closer to clinical application, we increased complexity of the sample in each step by using less depleted samples and faster preparative methods.…”

Proteomics biomarkers for solid tumors: Current status and future prospects

“…In combination with the serum markers CEA and cancer antigen 19‐9 (CA19‐9), an increased AUC value of 0.77 was reached ( P < 0.001) . Surinova et al performed proteomic profiling of microdissected human primary tumor epithelia compared to adjacent non‐malignant mucosa ( n = 16 each) using LC‐MS/MS to characterize tumor‐ associated secreted and cell surface glycoproteins . In total, 303 candidates were retrieved and further quantified initially in plasma samples from 19 CRC patients using selected reaction monitoring (SRM), as targeted proteomic approach.…”

“…In the first cohort consisting of 66 plasma samples from healthy controls, 34 patients with benign lesions and 100 CRC patients, a biomarker signature comprised of ceruloplasmin, serum paraoxonase/arylesterase 1 (PON1), serpin peptidase inhibitor clade A (SERPINA3), leucine‐rich alpha‐2‐glycoprotein (LRG1), and tissue inhibitor of metalloproteinases 1 (TIMP1) was established. Further validation of this biomarker signature in an independent set of plasma samples from 50 healthy controls, 17 benign lesions, and 202 CRC patients exhibited an AUC of 0.84 for the discrimination of CRC patients from the control groups …”

Clinical translation of MS-based, quantitative plasma proteomics: status, challenges, requirements, and potential

“…Using SIS peptides for normalization, the results revealed low ng/mL detection of 11 disease-related glycoproteins (from 12 N-linked glycopeptides), with a 4 peptide panel (from MEGF8, HSPG2, PRNP, and NCAM1) providing the diagnostic sensitivity (90.4%) and specificity (50.0%) necessary to screen for disease (n = means of detecting neurodegenerative disorders from plasma, as opposed to more invasive alternatives, such as cerebrospinal fluid (CSF, requires a lumbar puncture). Another promising finding involving N-glycan analysis came from a recent study by Aebersold R et al[77]. Their development revealed a 5-protein biomarker panel (includes CP, TIMP1, LRG1, PON1, and SERPINA3) that can sensitively detect and classify colorectal cancer in patient plasma samples.Further evaluation of the panel is, however, required to develop a fit-for-purpose clinical grade assay.…”