B-cell chronic lymphocytic leukemia is the most prevalent hematological malignancy in Western countries, accounting for approximately 11% of all malignant hemopathies and 25-30% of leukemias in adults. It is characterized by the accumulation of mature clonal B lymphocytes in the peripheral blood and in the bone marrow, which presents a high genomic heterogeneity and alterations in intracellular signalling pathways.
Multiple genetic alterations and mutations discovered in this pathology are reflected in the proteome of this cells. Therefore, the characterization of the proteome and their post-translational modifications can be a fundamental tool as a source of identification of possible therapeutic targets. To date, no studies have investigated so far the post-translational modification profile of B-CLL tumor cells and the correlation with the therapeucti algorithm.
For this reason, to improve the knowledge of the pathogenic mechanisms at the tumor cell level, analysis of quantitative protein levels and tyrosin-phospho profiles of B-CLL cells might be crucial for better mapping the altered signalling pathways leading to inappropriate proliferation and survival signalling in tumour cells and thereby, to better understanding and classification of the disease.
The main goal of this project is to design and develop a combine proteomics strategies based on PTMs analysis of tyrosine-phosphopeptides based on immunoenrichment and further LC-MS/MS analysis, in combination with the subsequent validation by employed a customized protein array and western blot (targeting tyrosine-kinases and BCR signalling pathways). Finally, all these data sets are integrated and correlated with the clinical-biological data. The study was performed in training cohort and valiated in a different cohort (100 samples)..
Our results provide new insights into the global proteome and phosphoproteome of B-CLL, immune signalling pathways involved in tumor development and progression at the tumor cell level; which are novel targets for immunotherapy has been identified and validated in a different cohort.