Přehled o publikaci

Recent studies show promising outcomes for venetoclax in the treatment of AML patients ineligible for intensive chemotherapy. Despite its success, up to one third of the patients do not respond to the therapy, and many experience disease relapse. One of the factors contributing to its lack of efficacy is alterations in the apoptotic pathway – mainly overexpression of MCL1 or BCL XL genes. Other factors and their interactions remain unresolved. To address this problem, our study aims towards revealing and understanding the key contributors to venetoclax resistance and finding novel molecular targets to combat the lack of treatment options. In this project, we generated venetoclax-resistant (VeR) AML cells derived from the following cell lines: MOLM-13, HL-60 and MV4-11. We then used RT-PCR and RNA sequencing to reveal the alterations caused by resistance generation. VeR cells were subjected to drug screening against a library of 859 approved drugs and the top performing drugs were further examined. In parallel, we performed genome-wide CRISPR/ Cas9 knockout screening using the Brunello CRISPR knockout library on both MOLM-13 wild-type and VeR cell lines. Results from the CRISPR screen revealed several genes whose loss sensitized the cells to venetoclax – MCL1, MARCH5, UBE2J2. We also identified genes whose deficiency may contribute to the venetoclax resistance – BAX, NOXA, ELAVL1, TP53. We further confirmed the deregulation in the apoptotic and metabolic pathway through RNAseq and identified DNA damaging agents, proteasome and HDAC inhibitors as the top candidates for all VeR cell lines via drug screening.