Project description:Chronic lymphocytic leukemia (CLL) is characterized by the accumulation of malignant CD5+ B lymphocytes (CLL cells) in the peripheral blood, and their progressive infiltration in lymphoid organs. MMP-9 plays an important role in cell migration and survival, contributes to CLL pathogenesis by proteolytic and non-proteolytic mechanisms and may constitutive a therapeutic target. We used Affimetrix microarray technology to characterize the global gene expression profile of chronic lymphocytic leukemia (CLL) cells upon MMP-9 transfection. The aim was to establish whether MMP-9 regulates gene expression and to identify new therapeutic targets in CLL.
Project description:To study the impact of SF3B1 mutations on alternative splicing and the effect of H3B-8800 splicing modulator in wild type and SF3B1-mutant chronic lymphocytic leukemia cells, we established SF3B1 K700E MEC1 CLL isogenic cell line and carried out RNA deep sequencing in SF3B1 wild type and K700E MEC1 cell lines upon H3B-8800 treatment.
Project description:SF3B1 mutations are recurrent genetic aberrations in chronic lymphocytic leukemia (CLL) that are particularly enriched in the clinically aggressive stereotyped subset #2. To elucidate the effect of SF3B1 mutations on splicing, we performed RNA-sequencing on 100 CLL subset cases, including 35 subset #2 cases. Zooming in on subset #2, we identified 61 differentially expressed splicing events when comparing 17 SF3B1mut to 18 SF3B1wt cases. A key event concerned the inclusion of an alternative exon in BRD9, a BAF chromatin remodeling complex component. We also detected alternative splicing in 5 additional transcripts related to the BAF complex: ZEB1, PLSCR1, PAPD5, DCAF16, and DLST. Long-read RNA-sequencing confirmed these alternative splice transcripts. In conclusion, spliceosome dysregulation caused by mutated SF3B1 leads to multiple splicing events and altered BAF complex function, highlighting a potential novel therapeutic vulnerability in SF3B1mut CLL.