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Truksa, J.

Publications and source records attributed to Truksa, J..

2 recordsLinked to original sources

Mitochondrially targeted deferasirox kills cancer cells via simultaneous iron deprivation and ferroptosis induction.

In principle, two separate ways to target cancer cells exist, the first one using iron deprivation and subsequent dysfunction of iron-dependent enzymes, and the second one inducing iron overload that leads to cell death known as ferroptosis. In this study, we introduce a compound that uniquely employs both pathways at once - mitochondrially targeted deferasirox (mitoDFX). MitoDFX deprives cells of biologically active iron while simultaneously depleting the primary cellular antioxidant glutathione (GSH) and inducing lipid peroxidation, both of which are hallmarks of ferroptosis. The role of the GSH is further supported by enhanced cell death in glutathione peroxidase 4 KO cells (GPX4 KO) induced by mitoDFX. This response is further exacerbated by simultaneous inhibition of glutathione metabolism or the pentose phosphate pathway. MitoDFX strongly affects the structure and function of mitochondria, leading to its fragmentation, ROS production and induction of mitophagy. Moreover, we found that mitoDFX treatment markedly reduces mitochondrial translation and downregulates levels of enzymatic subunits coupled with the ETC/TCA cycle as well as DNA replication and transcription, which could explain its strong cytostatic effects. In summary, mitoDFX achieves high activity and selectivity against cancer cells that seems unattainable for the conventional untargeted iron chelators, which have adverse effects on systemic iron metabolism. Our novel, targeted chelator has also shown enhanced efficacy in animal models, surpassing other mitochondrially targeted drugs and making mitoDFX a promising candidate for the development of future selective and highly effective cancer therapies.

cancer biology↗

Preparation of single-cell suspension from mouse breast cancer focusing on preservation of original cell state information and cell type composition

Single-cell analysis of gene expression has become a very popular method during the last decade. Unfortunately, appropriate standardization and workflow optimization remain elusive. The first step of the single cell analysis requires that the solid tissue be disassociated into a suspension of individual cells. However, during this step several technical bias can arise which can later result in the misinterpretation of the data. The goal of this study was to identify and quantify the effect of these technical factors on the quality of the single-cell suspension and the subsequent interpretation of the produced expression data. We tested the effects of various enzymes used for dissociation, several centrifugation forces, dissociation temperatures and the addition of Actinomycin D, a gene expression inhibitor. RT-qPCR was used to assess the effect from each parameter alteration, while a single-cell RNA sequencing experiment was used to confirm the optimized factors. Our concluding results provide a complete protocol for the tissue dissociation of mouse mammary tumour from 4T1 cells that preserves the original cell state and is suitable for any single-cell RNA sequencing analysis. Furthermore, our workflow may serve as a guide for the optimization of the dissociation procedure of any other tissue of interest, which would ultimately improve the reproducibility of the reported data.

cancer biology↗