Michał Seweryn Karbownik

Hyaluronan: towards novel anti-cancer therapeutics.

The understanding of the role of hyaluronan in physiology and various pathological conditions has changed since the complex nature of its synthesis, degradation and interactions with diverse binding proteins was revealed. Initially perceived only as an inert component of connective tissue, it is now known to be involved in multiple signaling pathways, including those involved in cancer pathogenesis and progression. Hyaluronan presents a mixture of various length polymer molecules from finely fragmented oligosaccharides, polymers intermediate in size, to huge aggregates of high molecular weight hyaluronan. While large molecules promote tissue integrity and quiescence, the generation of breakdown products enhances signaling transduction, contributing to the pro-oncogenic behavior of cancer cells. Low molecular weight hyaluronan has well-established angiogenic properties, while the smallest hyaluronan oligomers may counteract tumor development. These equivocal properties make the role of hyaluronan in cancer biology very complex. This review surveys recent data on hyaluronan biosynthesis, metabolism, and interactions with its binding proteins called hyaladherins (CD44, RHAMM), providing themolecular background underlying its differentiated biological activity. In particular, the article critically presents current ideas on actual role of hyaluronan in cancer. The paper additionally maps a path towards promising novel anti-cancer therapeutics which target hyaluronan metabolic enzymes and hyaladherins, and constitute hyaluronan-based drug delivery systems.

Board game versus lecture-based seminar in the teaching of pharmacology of antimicrobial drugs—a randomized controlled trial

The effectiveness of an educational board game developed to teach the pharmacology of antimicrobial drugs to medical students was compared with the lecture-based seminar as a supplemental tool to improve short- and long-term knowledge retention and the perception of the learning method by students. A group of 124 students was randomized to board game and control groups. Short-term knowledge retention was assessed by comparing differences in post- and pre-tests scores, and long-term knowledge retention by comparing final examination scores. Both didactic methods seem to improve short-term knowledge retention to similar extent. Long-term knowledge retention of board game seminar participants was higher than those who attended the lecture-based seminar (ANCOVA, P = 0.035). The effect was most pronounced within 14 days after the intervention (ANOVA, P = 0.007). The board game was well perceived by the students. The board game seems to be a promising didactic tool, however, it should be further tested to assess its full educational utility.

Polish individuals with an at-risk mental state: demographic and clinical characteristics.

The aim of this study is to present sociodemographic and clinical characteristics of Polish individuals with an at‐risk mental state (ARMS).

Antipsychotic Drugs Differentially Affect mRNA Expression of Genes Encoding the Neuregulin 1-Downstream ErbB4-PI3K Pathway

Background/Aims: The PIK3CD gene encodes the delta catalytic subunit of phosphoinositide 3-kinase (PI3K), an element of the neuregulin 1-downstream ErbB4-PI3K signaling pathway, which was recently identified as a molecular target for the treatment of schizophrenia. The aim of the study was to examine the effect of haloperidol (HALO), clozapine (CLO), olanzapine (OLA), quetiapine (QUE) and amisulpride (AMI) on the mRNA and protein expression of genes encoding the elements of ErbB4-PI3K pathway, in a human central nervous system cell line. Methods: The U-87MG human glioblastoma cell line was used as an experimental model. Quantitative polymerase chain reaction was used to examine the expression of mRNA and enzyme-linked immunosorbent assay for protein expression. Results: At concentrations reached in clinical settings in the brain, CLO, as well as OLA and QUE to a lesser extent, but not AMI and probably not HALO, decreased the mRNA expression of PIK3CD. Protein expression of the gene did not confirm the mRNA expression profile. Conclusions: The tested antipsychotic drugs (APDs) in the U-87MG glioblastoma cell line differentially regulates the mRNA expression of PIK3CD; however, the protein expression does not confirm these findings. The results of the study may help deepen the understanding of the mechanism of action of APDs.

Haloperidol, but not olanzapine, may affect expression of PER1 and CRY1 genes in human glioblastoma cell line

Abstract Background: There is barely any evidence of antipsychotic drugs affecting the molecular clockwork in human, yet it is suggested that clock genes are associated with dopaminergic transmission, i.e. the main target of this therapeutics. We decided to verify if haloperidol and olanzapine affect expression of CLOCK, BMAL1, PER1 and CRY1 in a human central nervous system cell line model. Methods: U-87MG human glioblastoma cell line was used as an experimental model. The cells were incubated with or without haloperidol and olanzapine in the concentration of 5 and 20 μM for 24 h. Real-time quantitative polymerase chain reaction with the ΔCT analysis was used to examine the effect of haloperidol and olanzapine on the mRNA expression of the genes. Results: At 5 μM, haloperidol decreased expression of CRY1 almost 20-fold. There was nearly a 1.5-fold increase in expression of PER1. Considering the 20 μM haloperidol concentration and both olanzapine concentrations, no other statistically significant effect was observed. Conclusions: At certain concentration, haloperidol seems to affect expression of particular clock genes in a human central nervous system cell line model, yet mechanism underlying this phenomenon remains elusive.

The ability of hyaluronan fragments to reverse the resistance of C6 rat glioma cell line to temozolomide and carmustine

Aim of the study Hyaluronan (HA) is an extracellular matrix (ECM) polymer that may contribute to the emergence of anti-cancer drug resistance. Attempts to reverse drug resistance using small hyaluronan oligomers (oHA) are being made. The initial reports suggest that the oHA fraction may effectively reverse anti-cancer drug resistance in glioma models. However, the reversal effects of oHA of defined molecular length on glioma cells have not been investigated yet. In this study, we examined HA fragments containing 2 disaccharide units (oHA-2), 5 disaccharide units (oHA-5), and 68 kDa hyaluronan polymer (HA-68k) as agents possibly reversing the resistance of a C6 rat glioma cell line to temozolomide (TMZ) and carmustine (BCNU). Material and methods A 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) viability assay was used to assess the cytotoxicity of TMZ and BCNU in the presence or absence of the hyaluronan fragments. By comparing viability of the cells, the reversal effects of HA fragments on TMZ and BCNU resistance in C6 glioma cells were assessed. Results We found statistically significant decreases in the viability of cells in the presence of TMZ+oHA-5 as compared to TMZ alone (51.2 ±4.5 vs. 74.2 ±5.8, p = 0.0031), BCNU+o-HA5 as compared to BCNU alone (49.3 ±4.4 vs. 65.6 ±5.7, p = 0.0119), and BCNU+HA-68k as compared to BCNU alone (55.2 ±2.3 vs. 65.6 ±5.7, p = 0.0496). Conclusions Conclusions: Hyaluronan oligomers of 5 disaccharide units (oHA-5) significantly reversed the resistance of C6 cells to TMZ and BCNU. The results are only preliminary and a more thorough follow-up investigation is required to assess their actual role.

Activation of phosphoinositide 3-kinase delta by antipsychotic drugs: Preliminary results

BACKGROUND Catalytic subunit delta of phosphoinositide 3-kinase, p110δ, encoded by the PIK3CD gene, was recently proposed as a target for pharmacological treatment of schizophrenia. Current antipsychotic drugs were found to decrease the mRNA expression of PIK3CD, but the mechanism of this process is not known. The aim of the study was to elucidate the mechanism by which antipsychotic drugs affect the mRNA expression of PIK3CD. METHODS The direct effect of haloperidol, clozapine, olanzapine, quetiapine and amisulpride on p110δ enzymatic activity was tested with a kinase assay, and the results were referenced against data on the mRNA expression of PIK3CD. RESULTS Haloperidol, clozapine, olanzapine and quetiapine, but not amisulpride, at the concentration of 20-80 μM, were found to significantly increase enzymatic activity of p110δ by up to two times in a dose-dependent manner. Linear regression analysis revealed that more than 40% of the variance in antipsychotic drugs-induced changes in the expression of PIK3CD mRNA was explained only by changes in antipsychotic drug-regulated p110δ enzymatic activity (p = 0.011). CONCLUSIONS Antipsychotic drugs differentially increase the enzymatic activity of p110δ. This effect is associated with that of mRNA expression of the PIK3CD gene. Drug-enzyme interaction may explain the effect of antipsychotic drugs on the expression of PIK3CD mRNA, however, further studies are needed to investigate this hypothesis.

miR-30a-5p together with miR-210-3p as a promising biomarker for non-small cell lung cancer: A preliminary study

BACKGROUND Although an immense effort has been made to develop novel diagnostic methods and treatment strategies for non-small cell lung cancer (NSCLC), the survival rate of this disease has remained virtually unchanged. Small non-coding RNAs called microRNAs (miRNAs) have appeared to be very promising biomarkers of cancer including NSCLC. OBJECTIVE We investigated the expression level of six miRNAs, and subsequently we evaluated their diagnostic ability and their clinical significance. METHODS We performed an analysis in 50 paired cancer and non-cancerous lung tissue samples collected from NSCLC patients. The RT-qPCR technique was used to investigate the expression profile. RESULTS Obtained results indicate that miR-30a-5p, miR-126-3p and miR-486-5p are downregulated, while miR-205-5p and miR-210-3p are upregulated in NSCLC tissue. Moreover, performed stepwise discriminant analysis determined the model including miR-30a-5p and miR-210-3p which tested on the test set (n= 30) revealed an AUC of 0.969 and provided 100% sensitivity and 80% specificity in discriminating NSCLC tissue from non-cancerous lung tissue. CONCLUSIONS The present preliminary study demonstrated that five tested miRNAs were deregulated in cancer tissue. Moreover, miR-30a-5p together with miR-210-3p with excellent sensitivity and acceptable specificity may distinguish cancer tissue form non-cancerous tissue and thus may become a potential diagnostic biomarker for NSCLC.