title: Toward personalized therapy for chronic lymphocytic leukemia: DSC and cDNA microarray assessment of two cases.
abstract: The differences in clinical course of chronic lymphocytic leukemia could have an impact on variations in a patient's response to therapy. Our published results revealed that thermal transition (95 ± 5°C) in differential scanning calorimetry profiles appear to be characteristic for the advanced stage of CLL. Moreover, a decrease/loss of this transition in nuclei from leukemic cells exposed to drugs ex vivo could indicate their diverse efficacy. It seems that the lack of changes in thermal profile could predict patient's drug resistance. In this study, we demonstrate the results obtained after drug treatment of leukemic cells by calorimetry, apoptosis-related parameters involved in expression of genes using cDNA microarray and western blot. These data were compared with the patients' clinical parameters before and after RCC therapy (rituximab + cladribine + cyclophosphamide). The complementary analysis of studied cases with opposite clinical response (CR or NR) revealed a strong relationship between clinical data, differences in thermal scans and apoptosis-related gene expression. We quantified expression of eight of apoptosis-related 89 genes, i.e., NOXA, PUMA, APAF1, ESRRBL1, CASP3, BCL2, BCL2A1 and MCL1. Particular differences in NOXA and BCL2 expression were revealed. NOXA expression in cells of patients who achieved a complete response to RCC therapy was 0.44 times higher in comparison to control ones. Interestingly, in the case of patients who did not respond to immunotherapy, NOXA expression was highly downregulated (RQ = 4.39) as compared with untreated cells. These results were confirmed by distinct cell viability, protein expression as well as by differences in calorimetry profiles.
authors.affiliation: Department of Cytobiochemistry, University of Lodz, Lodz, Poland.
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Rogalińska
authors.forename: Ida
authors.initials: I
authors.lastname: Franiak-Pietryga
authors.forename: Jerzy Z
authors.initials: JZ
authors.lastname: Błoński
authors.forename: Paweł
authors.initials: P
authors.lastname: Góralski
authors.forename: Henryk
authors.initials: H
authors.lastname: Maciejewski
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Janus
authors.forename: Paweł
authors.initials: P
authors.lastname: Robak
authors.forename: Marek
authors.initials: M
authors.lastname: Mirowski
authors.forename: Henryk
authors.initials: H
authors.lastname: Piekarski
authors.forename: Tadeusz
authors.initials: T
authors.lastname: Robak
authors.forename: Zofia M
authors.initials: ZM
authors.lastname: Kiliańska
mesh_terms: D000368:Aged; D058846:Antibodies, Monoclonal, Murine-Derived / Q000008:administration & dosage; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology / Q000627:therapeutic use*; D002152:Calorimetry, Differential Scanning; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D017338:Cladribine / Q000008:administration & dosage; D003520:Cyclophosphamide / Q000008:administration & dosage; D019008:Drug Resistance, Neoplasm; D006801:Humans; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D008297:Male; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D057285:Precision Medicine; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D000069283:Rituximab; D059467:Transcriptome; D016896:Treatment Outcome; D014407:Tumor Cells, Cultured
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; C498702:PMAIP1 protein, human; D019253:Proto-Oncogene Proteins c-bcl-2; D017338:Cladribine; D000069283:Rituximab; D003520:Cyclophosphamide