Project title: “Nanoparticles designed to target chemokine-related inflammatory processes in vascular diseases and cancer metastasis and implementation of a biosensor to diagnose these disorders”
Project type: European project – EuroNanoMed
An inflammatory process and the subsequent specific repertoire of chemokines and cell adhesion molecules expressed by activated vascular endothelial cells (EC) are common initial events in atherosclerotic plaque formation and cancer cell metastasis. By this process, particular immune cells (i.e. leukocytes) are attracted and participate to the developing atheroma and in cancer; tumour cells cross EC and metastasize within the tissues. In both cases, therapeutically blockage and/or early diagnosis (detection) of inflammation may prevent their dreadful outcome. The objectives of NANODIATER were: 1. to target specially designed nanoparticles (NP) carrying chemokines inhibitors that will be released restrictively at sites of activated endothelium in atherosclerotic plaque formation and tumour cell metastasis and 2. to design and develop a NP-derived diagnostic kit (“cell sensor”) for non-invasive detection of blood circulating inflammatory cells or metastatic tumour cells that can predict the evolution of these diseases.
For objective 1, two types of targeted NP encapsulating chemokine antagonists (CA) or chemokine receptor antagonists (CRA) were prepared: stabilised target-sensitive liposomes (TSL) and polymeric nanoparticles (PNP) that had attached on their surface monoclonal antibodies or peptides that recognize VCAM-1 expressed by activated EC. The specific binding and fate of targeted NP and the efficiency of CA and CRA released were tested using in vitro and in vivo experiments employing different cultured activated EC (under static and dynamic conditions) and assessing the effect on the decrease of monocyte or tumour cell adhesion/migration. For in vivo testing, animal model of atherosclerosis (ApoE-deficient mice) and cancer (mice with metastatic seeding of tumour cells or metastasis of tumour cells with chemokine knock-down, and patient-derived tumour models) were employed. Based on the results, from all targeted NP the best ones for preclinical characterization were selected.
For objective 2, a cell sensor using metallic NP conjugated to antibodies directed towards specific cellular markers was designed. The cells (leukocytes or cancer cells) were incubated with the biosensor and an electrochemical multiplexed immunoassay protocol for simultaneous measurements of different cellular markers was developed. The biosensor was tested on the blood of mice. The correlation between the number of detected cells and the degree of metastasis or atherosclerosis was determined.