Multiplexed Detection of Pancreatic Cancer by Combining a Nanoparticle-Enabled Blood Test and Plasma Levels of Acute-Phase Proteins

Abstract

The development of new tools for the early detection of pancreatic ductal adenocarcinoma (PDAC) represents an area of intense research. Recently, the concept has emerged that multiplexed detection of different signatures from a single biospecimen (e.g., saliva, blood, etc.) may exhibit better diagnostic capability than single biomarkers. In this work, we develop a multiplexed strategy for detecting PDAC by combining characterization of the nanoparticle (NP)-protein corona, i.e., the protein layer that surrounds NPs upon exposure to biological fluids and circulating levels of plasma proteins belonging to the acute phase protein (APPs) family. As a first step, we developed a nanoparticle-enabled blood (NEB) test that employed 600 nm graphene oxide (GO) nanosheets and human plasma (HP) (5% vol/vol) to produce 75 personalized protein coronas (25 from healthy subjects and 50 from PDAC patients). Isolation and characterization of protein corona patterns by 1-dimensional (1D) SDS-PAGE identified significant differences in the abundance of low-molecular-weight corona proteins (20–30 kDa) between healthy subjects and PDAC patients. Coupling the outcomes of the NEB test with the circulating levels of alpha 2 globulins, we detected PDAC with a global capacity of 83.3%. Notably, a version of the multiplexed detection strategy run on sex-disaggregated data provided substantially better classification accuracy for men (93.1% vs. 77.8%). Nanoliquid chromatography tandem mass spectrometry (nano-LC MS/MS) experiments allowed to correlate PDAC with an altered enrichment of Apolipoprotein A-I, Apolipoprotein D, Complement factor D, Alpha-1-antichymotrypsin and Alpha-1-antitrypsin in the personalized protein corona. Moreover, other significant changes in the protein corona of PDAC patients were found. Overall, the developed multiplexed strategy is a valid tool for PDAC detection and paves the way for the identification of new potential PDAC biomarkers.

Inflammatory biomarkers and nanotechnology: new insights in pancreatic cancer early detection

“…Recently, the diagnostic ability for PDAC has been improved by combing the PC analysis with non-specific laboratory data such as haemoglobin or acute phase protein levels. 20 , 21 This novel approach demonstrated high discrimination between PDAC and healthy subjects in up to 90% of cases, thus appropriate for PDAC diagnosis.…”

Stratifying Risk for Pancreatic Cancer by Multiplexed Blood Test

“…Cyto-histologically proven PDAC patients admitted to the Fondazione Policlinico Universitario Campus Bio-Medico that met the inclusion criteria reported in [35] have been considered eligible for the analysis. Their clinical and laboratory tests data were collected.…”

Coupling magnetic levitation of graphene oxide–protein complexes with blood levels of glucose for early detection of pancreatic adenocarcinoma

“…Recently, our group has demonstrated that a proper combination of non-specific laboratory data (e.g., low hemoglobin levels and acute phase protein concentration), with the outcomes of nanoparticle-enabled blood (NEB) tests, can discriminate PDAC patients from healthy controls with high sensitivity, specificity, and diagnostic accuracy. The NEB tests assume that the protein corona that forms around nanoparticles (NPs) upon exposure to human plasma (HP) is personalized and, under certain experimental conditions, contains a signature for PDAC (Caputo et al 2020;Caputo et al 2022a). The most recent version of the NEB test is based on the analysis of the magnetic levitation (MagLev) profiles of coronated graphene oxide (GO) nanoflakes in a paramagnetic solution (Digiacomo et al 2021;Digiacomo et al 2022;Quagliarini et al 2022a).…”