Project description:Based on previous work on the KDM histone demethylase family in AML, the transcription factor NFATC2 was identified as a regulatory target of KDM4A in MLL-AF9-rearrangement THP-1 cells. Our existing work examined the elements of the transcriptome regulated by NFATC2 in THP-1 cells and so, to further this work, we aimed to characterise the DNA-binding targets of NFATc2 in these cells. In this study, we used NFATc2 immunoprecipitation for ChIP-seq in untreated THP-1 cells, in order to determine these.
Project description:Analysis of the transcriptome of THP-1 cells upon Huh7 cell-derived ectosomes incubation. Monocytic THP-1 cells were incubated with or without ectosomes (10 mg/ml) derived from scramble sequence transfection (Scramble Ecto) or PKM2 knocking down (PKM2 KD Ecto) Huh7 cells for 24 h. Results provide a novel insight into monocyte differentiation.
Project description:Allele-specific regulatory events are essential in many fundamental biological processes. In this study, we utilized the human monocytic cell line THP-1 and performed RNA immunoprecipitation sequencing (MeRIP-seq) to explore allele-specific regulation of N6-methyladenosine (m6A) RNA modifications. Recent research highlights the importance of allele-specific regulation of m6A (ASm6A) and the need for precise detection methods to elucidate its underlying mechanisms. To address limitations in current approaches, which often emphasize genetic variations rather than comprehensive regulatory mechanisms, we applied M6Allele, a meta-analysis framework based on hierarchical Bayesian models. This method enables accurate detection of ASm6A events at the peak level, providing new insights into allele-specific regulatory mechanisms of m6A modifications. Our findings from the THP-1 cell line contribute to a deeper understanding of ASm6A and its biological significance.
Project description:The hypomethylating agent 5-azacytidine (AZA) is the first-line induction therapy for AML patients unsuitable for intensive chemotherapy. The anti-tumor effect of AZA results in part from T-cell cytotoxic responses against MHC-I-associated peptides (MAPs) deriving from hypermethylated genomic regions such as cancer-testis antigens (CTAs), or endogenous retroelements (EREs). However, clear evidence supporting higher ERE MAPs presentation after AZA treatment in AML is lacking. Interestingly, we find that AZA-mediated DNMT2 inhibition leads to autophagy induction, which is responsible for mitigating ERE MAPs generation. To validate our findings, we examined the immunopeptidome of THP-1 cells treated with AZA combined with autophagy inhibitor, Spautin-1 or decitabine (DAC, non-DNMT2 inhibiting HMA) through a proteogenomic approach.
Project description:There is currently no human macrophage cell line and in vitro experiments requiring these cells currently require mitogenic stimulation of a macrophage precursor cell line (for example THP-1 cells) or ex vivo maturation of circulating primary monocytes. We have derived a unique sub-clone of the THP-1 cell line capable of spontaneous and perpetual differentiation into alveolar macrophage-like cells. Here we used the Agilent G4851C (v3) array to look at the gene expression of the new cell line which can be compared it with THP-1 cells and PMA stimulate THP-1 cells (Accession number E-MTAB-4153). In addition the new human alveolar macrophage like cell line has been treated with unmodified lipid to look at lipid uptake by these cells and the effect on gene expression.
Project description:Bortezomib is a proteasome inhibitor used in severel different hematological malignancies. Resistance to this drug is still poorly understood. In order get more insight in the resistance mechanism, we developed several bortezomib resistant subclones of the THP-1 monocytic/macrophage cell line. On these subclones expression arrays were performed. We performed expression array three different bortezomib resistant subclones of the THP-1 cell line. The resistant subclones were spotted against the parental THP-1 wildtype cell line.