Chronic Lymphocytic Leukemia (CLL) represents a 30% of leukemia cases in Europe. Despite the immense progress to understand the first oncogenic events that lead the apparition of the disease, the way protein pathways are involved in the pathogenesis of CLL is still unknown. The main goal of the project is to understand which mechanisms at protein level make differences at clinical level in CLL patients. A better knowledge of the different protein pathways will be useful for the discovery of new biomarkers for early diagnostic and personalized medicine. It is proposed to apply a combination of high-throughput methodologies coupled to label-free and labelled detection methods. The project promoter University of Salamanca is focused on the relationship between the immune response and cancer, as well as on the study of malignancies derived from cells of the immune system. The donor partner Oslo University Hospital is trying to make proteomics simple and accessible. The main beneficiaries of the project will be the scientific community and citizens at all levels.
Summary of project results
The interactome of CLL cells is largely uncharacterized. The project opens a new hallmark for a network of cooperation between group leaders in the field; because of a synergy between both labs due to the combination of these groups are able to overcome a project which requires a novel methodology development and biomedical research project. The current project could help generate new applications that would be rapidly introduced in clinical routine for diagnostic and follow-up of these haematological malignancies. The main goal was to deciphering the cell signalling pathways involved in the LLC pathology, and the differential protein expression profiles between aberrant and normal cells. A novel methodology for analysis of proteins in high-throughput format, based on the combination of Size Exclusion Chromatography (SEC) and the antibody arrays on microspheres format (Microsphere Affinity Proteomics, MAP). During the project, the methodology has been implemented in both participating labs. The methodology has four steps: subcellular protein extraction; biotinylation of cell lysates; protein separation by FPLC, and protein analysis by antibody coupled to color-coded beads. Finally, the colour coded and the targeted proteins (labelled with streptavidin) are detected by flow-cytometry. In addition, SEC-MAP methodology also allows the subcellular localization of the protein and the role in protein complexes. Additionally, the expression (in cytoplasm and membrane) of the several BCR molecules was analysed, by flow-cytometry. In the study, a total of 79 samples of peripheral blood of patients of B-CLL from hematology Department at Hospital Clínico Universitario of Salamanca have been included, Also, 4 samples of healthy donors, obtained from the Centro de Hemodonación y Hemoterapia de Castilla León. During the project implementation, several members of the involved research teams have participated in events such as the Congress of Iberian Cytometry Society. At the closure of the project funding, two publications were in preparation to be submitted during 2016.
Summary of bilateral results
The project opened a direct collaboration between two leading groups in the area, allowing a synergy helping advance in the knowledge of the analyzed pathology. As a result, both involved labs are working together with the main goal of analyzing the data obtained and in generating publications. The project relevance can be described in two approaches. On the one side, biomedical research, which has a strong influence in the translational clinic due to the discoveries that could be applied in treatment and diagnosis. On the other side, the project is highly relevant from the methodological point of view, as it is based on a novel approach that could be protected by patent (potential interest: biotechnological and pharmaceutical companies).