Project description:To clarify the role of DMD in macrophages in response to IMQ, we performed transcriptomic analysis by a Low Input Quick-Amp Labeling kit (Agilent Technologies, USA) from IMQ-induced THP1 macrophages with or without DMD treatment in vitro.
Project description:Macrophages are key cell types of the innate immune system regulating host defense, inflammation, tissue homeostasis and cancer. Within this functional spectrum diverse and often opposing phenotypes are displayed which are dictated by environmental clues and depend on highly plastic transcriptional programs. Among these the ‘classical’ (M1) and ‘alternative’ (M2) macrophage polarization phenotypes are the best characterized. Understanding macrophage polarization in humans may reveal novel therapeutic intervention possibilities for chronic inflammation, wound healing and cancer. Systematic loss of function screening in human primary macrophages is limited due to lack of robust gene delivery methods and limited sample availability. To overcome these hurdles we developed cell-autonomous assays using the THP-1 cell line allowing genetic screens for human macrophage phenotypes. To confirm the relevance of the THP1-based system we performed microarray studies on THP1 cells and primary human cells subjected to various conditions. THP1 cells were treated with PMA (phorbol 12-myristate 13-acetate) to derive macrophages which were then cultured 1, 3 or 6 days with different stimuli.
Project description:We then performed gene expression profiling analysis using data obtained from RNA-seq of 9 skin tissues including 3 per group in vaseline-treated group, imq model group and bergapten-gel-treated imq model group.
Project description:To explore the detailed mechanism by which hsa_circ_0004287 inhibits M1 macrophage activation, we performed RNA-seq in control and hsa_circ_0004287 knockdown THP1-derived macrophages, and identified 43 upregulated and 35 downregulated genes with the criterion of |FC|≥2 and p≤0.05.
Project description:We delved into the consequences of macrophages that co-cultured with PDAC cells with Vector/MLKL overexpression. We treated THP1 derived macrophages with PANC1 cells with Vector/MLKL overexpression.
Project description:DMD is a genetic disease, which leads to muscle weakness and cardiomyopathy. The latter remains incurable, being the main cause of death in DMD, therefore new therapeutic strategies are being sought to provide effective treatment. One of them considers upregulation of utrophin, a protein structurally and functionally homologous to dystrophin. In this study proteomic analysis of dystrophin-deficient and both dystrophin- and utrophin-deficient hiPSC-CM indicated on considerable differences in terms of contraction-related mechanisms. We thus investigated the role of utrophin in the maintenance of electrophysiological properties of DMD hiPSC-CM using the cells with additional utrophin deficiency and with utrophin upregulation. Obtained results indicated on disturbance of calcium handling in DMD hiPSC-CM, even more pronounced in DMD/UTRN KO hiPSC-CM and increased values of AHP in DMD hiPSC-CM. Utrophin upregulation improved both calcium oscillations and AHP values. Our findings highlight utrophin as important in the maintenance of the electrophysiological properties of DMD hiPSC-CM.