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HPLC-based Assay to Monitor Extracellular Nucleotide/Nucleoside Metabolism in Human Chronic Lymphocytic Leukemia Cells
Published on: July 20, 2016
Sensitivity of chronic lymphocytic leukemia cells to small targeted therapeutic molecules: An in vitro comparative
Sandra Eketorp Sylvan1, Henriette Skribek2, Stefan Norin3
1Department of Hematology, Karolinska University Hospital, Stockholm, Sweden; Department of Oncology-Pathology, Karolinska Institutet, Stockholm, Sweden.
Abstract:
New drugs targeting important cellular signaling pathways are currently being developed for chronic lymphocytic leukemia (CLL). It is therefore of interest to analyze their in vitro killing capacity in manufacturer-independent, comparative experiments. We here report on the sensitivity of CLL cells to a panel of emerging targeted therapeutics using high-throughput screening based on an automated fluorescence digital scanning system. Fresh CLL cells from 42 patients with indolent or progressive CLL were cultured for 72 hours on microtiter plates in a unique primary cell culture medium. Antitumor effects of 31 small therapeutic molecules (and, as controls, 29 cytostatic agents) at equimolar concentration were compared in a fluorescence survival assay. In vitro sensitivity to each drug exhibited considerable interpatient variability. The highest mean direct killing was observed for one survivin inhibitor (YM-155), two bcl-2 inhibitors (ABT-199, ABT-737), and one selective CDK inhibitor (dinaciclib). Their killing capacity was, in contrast to most cytostatic agents, similarly high in refractory versus untreated CLL patients and was significantly higher on cells with the 17p deletion/TP53 mutation than on cells with other cytogenetic abnormalities (p = 0.02). Sensitivity of bone marrow and lymph node cells was highly correlated with that of blood cells. Even though direct killing may not be the only therapeutic effector function in vivo, results from this head-to-head comparison may help to identify drugs of particular interest for intensified clinical development.
Insights
New targeted therapies show promise for chronic lymphocytic leukemia (CLL). Bcl-2 and survivin inhibitors demonstrated high in vitro killing capacity, especially in patients with specific genetic mutations.
Area of Science:
- Oncology
- Pharmacology
- Molecular Biology
Background:
- Chronic lymphocytic leukemia (CLL) treatment is evolving with new targeted drugs.
- Evaluating drug efficacy in manufacturer-independent comparative studies is crucial.
Purpose of the Study:
- To compare the in vitro killing capacity of emerging targeted therapeutics against CLL cells.
- To identify drugs with significant antitumor effects in a head-to-head screening.
Main Methods:
- High-throughput screening of 31 small therapeutic molecules and 29 cytostatic agents on fresh CLL cells from 42 patients.
- Utilized an automated fluorescence digital scanning system and a unique primary cell culture medium for 72-hour cultures.
- Assessed antitumor effects at equimolar concentrations in a fluorescence survival assay.
Main Results:
- Considerable interpatient variability in drug sensitivity was observed.
- Survivin inhibitor YM-155, bcl-2 inhibitors ABT-199 and ABT-737, and CDK inhibitor dinaciclib showed the highest mean direct killing.
- These drugs were effective in refractory CLL and particularly potent against cells with 17p deletion/TP53 mutation.
Conclusions:
- Emerging targeted drugs, especially bcl-2 and survivin inhibitors, exhibit potent in vitro killing of CLL cells.
- Drug sensitivity varied based on patient genetics, with 17p deletion/TP53 mutation conferring higher sensitivity.
- These findings support further clinical development of identified high-efficacy drugs for CLL.
