Identification and priming of large diffuse B-cell lymphomas dependent on the signal from the B-cell receptor. From tools of large-scale data analysis to the development of rational anti-cancer therapies

The aim of the project is to identify and stratify Diffuse large D-cdl lymphoma (DLBCL) signal-dependent B-cell receptor (BCR) with low or high activity of the transcription factor NF- KB. As part of the project, it is planned to study the anti-tumor activity of selected BCR signal inhibitors in DLBCL cell lines and primary DLBCL cells, taking into account their division into BCR-dependent and BCR-independent lines as well as with low and high basal activity of the NF-KB transcription factor. At a later stage of the project, it is also planned to develop a comprehensive antibody panel to analyze the signal status of BCR and the response to therapy with BCR inhibitors in established and primary DLBCL cell lines by mass cytometry - CyTOF. In parallel, comprehensive characterization of genetic changes in the BCR signaling pathway in the model of 317 primary DLBCL cell lines with known cell-of-origin (COO) and consensus cluster classification (CCC) status, subjected to exome sequencing, will be carried out. DLBCL is an aggressive lymphoma with a diverse genetic background, and thus - with a varied clinical picture. The project involving research into the molecular mechanisms of DLBCL pathogenesis related to signaling with BCR is important because BCR signal inhibitors have already been registered for the treatment of chronic lymphocytic leukemia (CLL), but not DLBCL, although both diseases are dependent on the signal from the BCR. In DLBCL, morphologically identical cells may rely on completely different pro-life signaling pathways and thus respond differently to chemical inhibition of BCR-associated kinases. A better understanding of the mechanisms leading to neoplastic transformation of DLBCL B cells could contribute to a better understanding of the anti-tumor activity of BCR inhibitors. The effect of inhibitors of BCR-related proximal kinases on DLBCL cells dependent on this receptor signal has not been systematically studied so far. Despite many reports indicating that BCR inhibitors have antitumor effect, it has not been clearly established what the mechanism of their action is and whether there are interactions between various drugs in this group. Preliminary results indicate that a combination of different BCR inhibitors should be used to achieve a more effective anti-tumor activity in DLBCL therapy. Such a strategy would primarily prevent the activation of pro-life compensatory pathways in neoplastic cells. However, this issue requires detailed research, which is planned to be carried out under this project. Despite significant advances in cancer genomics in recent years and the publication of numerous results from the sequencing of the genomes of cells of many types of cancer, genetic data on the primary clinical material from DLBCL patients is still insufficient to identify with a high probability the mutations that play a leading role in pathogenesis of this disease. The expected result of the proposed project will be: a) Identification of the most therapeutically beneficial combinations of BCR inhibitors depending on the molecular basis of DLBCL. b) Development of a comprehensive and convenient CyTOF antibody panel for DLBCL molecular substrate analysis and cell responses to therapy with BCR inhibitors. Analysis of the sequencing data of the exomic pool of 317 primary DLBCL cells to search for new mutations of potential prognostic and therapeutic importance in this disease.