Session: 605. Molecular Pharmacology and Drug Resistance: Lymphoid Neoplasms: Poster I
Hematology Disease Topics & Pathways:
Research, Fundamental Science, apoptosis, adult, Translational Research, drug development, cell expansion, Therapies, Biological Processes, Technology and Procedures, Study Population, Human
Our high-throughput screen of 2,393 compounds from the FDA library identified 33 drug candidates which substantially reduced the viability of MWCL-1 cells as compared to vehicle control (dimethyl sulfoxide). This screen yielded a hit rate of 1.3%, based on the cut-off score of IC50 2 μM or less. Of these hits, we identified five highly potent compounds with an IC50 of less than 0.1 μM (Fig. 1A). We confirmed the cytotoxicity potential of these lead compounds and found that all five compounds reduced the viability of MWCL-1 cells as early as 12 hours after treatment, with the highest reduction in viability observed at 72 hours. Furthermore, these compounds substantially reduced the viability of another WM cell line called BCWM.1 which morphologically and phenotypically resembles lymphoplasmacytic cells of WM. Importantly, we observed that these compounds reduced the viability of bone marrow-derived primary lymphocytes from a patient with WM as compared to vehicle control (Fig. 1B). The compound SN01005979 was most effective and exerted a similar cytotoxic potential as standard treatment Doxorubicin (Fig. 1B). To characterize the mechanisms of action by which these lead compounds kill MWCL-1 and BCWM.1 cells, we performed immunoblotting and tested the activation of programmed cell death pathways including caspase-1, gasdermin (GSDM)-D, GSDME (pyroptosis), caspase-8 (extrinsic apoptosis), caspase-9 (intrinsic apoptosis), caspase-3, -6, and -7 (executioners of apoptosis), and phosphorylation of MLKL (necroptosis). We found that these compounds selectively induce caspase-3-dependent cleavage of GSDME.
In summary, using a high throughput screen, we identified novel drug candidates with potent cytotoxic activity against WM. Our identification of key molecules targeted by these drug compounds could pave the way for the treatment of tumours expressing caspase-3 and GSDME. Moreover, our results offer new insights into the mechanism of certain FDA-approved drugs and could help in the development of safer and more effective cancer chemotherapy against WM and potentially other cancers. Future studies are required to test the tumour-killing potential of these drugs in vivo using mouse and humanised mouse models of WM.
Disclosures: Talaulikar: Amgen: Honoraria; Beigene: Honoraria; Takeda: Honoraria; CSL: Honoraria, Speakers Bureau; EUSA: Honoraria; Antengene: Honoraria; Roche: Honoraria, Research Funding, Speakers Bureau; Janssen: Honoraria, Research Funding, Speakers Bureau.
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