In medicine there is always an incentive to personalize therapy to lower side effects, improve
treatment outcomes and reduce the strain on the patient. In patients with chronic lymphocytic
leukemia (CLL) the adjustment of the treatment is preferable, as CLL is a heterogeneous and
age-related disease. In the presence of comorbidity and impairment of some of the bodily
functions in elderly population, tailored therapy is preferable, but with in vitro tests it is
difficult to predict an in vivo response. Many high throughput methods are being developed to
determine the ex vivo cell response and evaluate the cytotoxicity of active substances.
This master’s thesis aims to examine a new approach to the treatment of a patient with CLL
with tailored therapy, with optimization of HexascopeHEAMA system. We analyzed 48 blood
samples of patients with CLL. Peripheral mononuclear blood cells were firstly isolated, then
they were incubated in presence of 30 different active substances with different
concentrations. To optimize the method, the influence of the incubation time and the final
concentration of cells on the microtiter plate were checked and the reproducibility of the test
was evaluated. For the clinical part of the study, 35 samples were selected from these 48 blood
samples. Data on genetic mutations, disease stage and therapies received were found in the
internal database of UMC Ljubljana. Based on the collected data, it was possible to compare
and evaluate the analysis that were carried out.
HexascopeHEAMA cytotoxicity test showed that, for patients with CLL with adequate
concentration of cells on the microtiter plate and sufficient infiltration of lymphocytes in the
blood, the results were in compliance with quality requirements and reproducibility. The
prediction of an ex vivo response with in vivo efficacy was not successful, as data on the actual
outcome of the patient's treatment were lacking. A weak point in performing the test were also
the active substances. Patients with CLL are often treated with a combination of active
substances, which was something impossible to test with the used system. In order to
introduce the system into everyday practice, it might be reasonable to consider changing the
set of active substances or rather, to introduce combinations (therapeutic schemes) of active
substances with which patients are treated. Due to the fact that monoclonal antibodies are
generally used in the treatment of CLL, the HexascopeHEAMA system has limited utility in
patients with CLL, because it does not include them.
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