Personalized therapy tests for the monitoring of chronic lymphocytic leukemia development

Abstract

There is individual variation in the course of disease development and response to therapy of patients with chronic lymphocytic leukemia (CLL). Novel treatment options for CLL include a new generation of purine analogs, antibodies and inhibitors of specific cell signaling pathways, which typically induce apoptosis or necrosis. A prospective analysis of patient blood samples revealed that a combination of four tests allowed the most appropriate and effective type of treatment to be selected prior to drug administration, and for the analysis of leukemic cell sensitivity to anticancer drug(s) during disease development. The comparative analysis of blood from the stable and progressive form of CLL in an individual patient revealed diversity in the response to anticancer agents. CLL peripheral blood mononuclear cells were incubated with cladribine + mafosfamide (CM), fludarabine + mafosfamide, CM + rituximab, rituximab alone (Rit) or kinetin riboside (RK). A combination of cell viability, differential scanning calorimetry (DSC) profiles of nuclear preparations and poly(ADP-ribose) polymerase 1 (PARP-1) protein expression analysis of the leukemic cells was performed to evaluate the anticancer effects of the tested agents during CLL development. The results of the present study indicate that such studies are effective in determining the most appropriate anticancer drug and could monitor disease progression on an individual level. In addition, the results of the current study suggest that CLL progression leads to diversification of the cellular drug response. The most efficient apoptosis inducer for the patient was purine analog RK when the disease was stable, while the CM combination was the most effective agent for the progressive form of disease.

Dose and drug changes in chronic lymphocytic leukemia cell response in�vitro: A comparison of standard therapy regimens with two novel cyclin‑dependent kinase inhibitors

“…Our findings reveal significant differences in the response to anticancer agents among leukemic patients, as noticed in previous studies (8,14,15). Moreover, our results show that the comparison of cell viability, apoptosis rate, and the level of necrosis underline the importance of proper anticancer agent dose selection to avoid the occurrence of a high necrosis level that could transform in vivo into inflammation.…”

“…It must be stated that these combined results are difficult to compare with those of each single patient. It is important to note that the results for patients 15, 16 or 17 presented here indicate that the median results presented in scientific papers usually provide incorrect recommendations (13–15) concerning patient resistance/sensitivity to anticancer agents, and these would not be effective in treating patient no. 16.…”

“…The representative results for patients (3 presented as single cases and 2 combined figures with statistics) were included in current manuscript, showing distinct patients response to anticancer agents. The effect of CDK inhibitors on CLL cells were analysed for 6 CLL patients and compared with standard chemotherapeutic regimens (CM, cladribine + mafosfamide; RCM, rituximab + cladribine + mafosfamide), as described in our previous studies with more than 50 patients (14,15). The CLL cells isolated from peripheral blood of patients were incubated with anticancer agents for 24 and 48 h, harvested and then analyzed for viability, apoptosis and necrosis by flow cytometry.…”

“…Since the disease could transform into an active form, the coexistence of quiescent and cycling cell populations complicate the diagnostics and prediction of therapy efficacy (5,6). Therefore, an analysis of the chemosensitivity profile of patient cells prior to therapy followed by the use of personalized medicine based on apoptosis induction would markedly reduce the ineffective treatment (8,13–15).…”

“…Cell viability, the rate of apoptosis and necrosis were estimated using a Vybrant Apoptosis Assay #4 kit after 0, 24 and 48 h, as in previous studies (14,15). The early apoptotic cells were marked with YO-PRO fluorescent dye, and late apoptotic and necrotic cells with propidium iodide.…”