The placental DNA methylation landscape is unique, with widespread partially methylated domains (PMDs). The placental "methylome" is conserved across mammals, a shared feature of many cancers, and extensively studied for links with pregnancy complications. Human trophoblast stem cells (hTSCs) offer exciting potential for functional studies to better understand this epigenetic feature; however, whether the hTSC epigenome recapitulates primary trophoblast remains unclear. We find that hTSCs exhibit an atypical methylome compared with trophectoderm and 1st trimester cytotrophoblast. Regardless of cell origin, oxygen levels, or culture conditions, hTSCs show localized DNA methylation within transcribed gene bodies and a complete loss of PMDs. Unlike early human trophoblasts, hTSCs display a notable absence of DNMT3L expression, which is necessary for PMD establishment in mouse trophoblasts. Remarkably, we demonstrate that ectopic expression of DNMT3L in hTSCs restores placental PMDs, supporting a conserved role for DNMT3L in de novo methylation in trophoblast development in human embryogenesis.

• MeSH
• DNA (Cytosine-5-)-Methyltransferases * metabolism   genetics   MeSH
• Epigenome MeSH
• Stem Cells metabolism   cytology   MeSH
• Humans MeSH
• DNA Methylation * genetics   MeSH
• Mice MeSH
• Placenta * metabolism   cytology   MeSH
• Pregnancy MeSH
• Trophoblasts * metabolism   cytology   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Pregnancy MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Pegylated interferon alfa (pegIFN-α) can induce molecular remissions in patients with JAK2-V617F-positive myeloproliferative neoplasms (MPNs) by targeting long-term hematopoietic stem cells (LT-HSCs). Additional somatic mutations in genes regulating LT-HSC self-renewal, such as DNMT3A, have been reported to have poorer responses to pegIFN-α. We investigated whether DNMT3A loss leads to alterations in JAK2-V617F LT-HSC functions conferring resistance to pegIFN-α treatment in a mouse model of MPN and in hematopoietic progenitors from patients with MPN. Long-term treatment with pegIFN-α normalized blood parameters and reduced splenomegaly and JAK2-V617F chimerism in single-mutant JAK2-V617F (VF) mice. However, pegIFN-α in VF;Dnmt3aΔ/Δ (VF;DmΔ/Δ) mice worsened splenomegaly and failed to reduce JAK2-V617F chimerism. Furthermore, LT-HSCs from VF;DmΔ/Δ mice compared with VF were less prone to accumulate DNA damage and exit dormancy upon pegIFN-α treatment. RNA sequencing showed that IFN-α induced stronger upregulation of inflammatory pathways in LT-HSCs from VF;DmΔ/Δ than from VF mice, indicating that the resistance of VF;DmΔ/Δ LT-HSC was not due to failure in IFN-α signaling. Transplantations of bone marrow from pegIFN-α-treated VF;DmΔ/Δ mice gave rise to more aggressive disease in secondary and tertiary recipients. Liquid cultures of hematopoietic progenitors from patients with MPN with JAK2-V617F and DNMT3A mutation showed increased percentages of JAK2-V617F-positive colonies upon IFN-α exposure, whereas in patients with JAK2-V617F alone, the percentages of JAK2-V617F-positive colonies decreased or remained unchanged. PegIFN-α combined with 5-azacytidine only partially overcame resistance in VF;DmΔ/Δ mice. However, this combination strongly decreased the JAK2-mutant allele burden in mice carrying VF mutation only, showing potential to inflict substantial damage preferentially to the JAK2-mutant clone.

• MeSH
• Cell Self Renewal MeSH
• Drug Resistance, Neoplasm * genetics   MeSH
• DNA Methyltransferase 3A * genetics   MeSH
• DNA (Cytosine-5-)-Methyltransferases * genetics   metabolism   MeSH
• Hematopoietic Stem Cells * metabolism   pathology   drug effects   MeSH
• Interferon-alpha * pharmacology   MeSH
• Janus Kinase 2 * genetics   metabolism   MeSH
• Humans MeSH
• Myeloproliferative Disorders * genetics   pathology   drug therapy   metabolism   MeSH
• Mice, Inbred C57BL MeSH
• Mice MeSH
• Polyethylene Glycols pharmacology   MeSH
• Recombinant Proteins MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

AIMS: The aim of this study was to compare the expression profile of selected DNA methyltransferases and global DNA methylation status in patients with different phases of multiple myeloma (MM) . For the analysis, different cellular populations including unsorted myeloma cells and a set of plasma cells purified by relevant antibodies were used. Consequently, laboratory data were compared to patients' clinical data. PATIENTS AND METHODS: For the analysis, unsorted bone marrow cell population of 44 MM patients (30 newly diagnosed, 9 relapsed and 5 patients in remission) and a set of 8 patients' samples of sorted plasma cells were used. We used commercially available RNA isolated from BM of 3 healthy individuals as control samples. Expression analysis of three DNA methyltransferases - DNMT1, DNMT3A, and DNMT3B was performed by quantitative RT-PCR and the patient global DNA methylation profiles were detected by colorimetric assay. RESULTS: Unchanged DNMT1 expression was detected in the selected cohort of patients. Normalized DNMT3A gene expression was globally higher in comparison with controls in unsorted and sorted cell populations. Low (0.08-1.81%) global DNA methylation status in unsorted samples of multiple myeloma patients did not correlate either with expression profiles of monitored DNA methyltransferases or with the stages of MM based on Durie-Salmon and International Staging System. CONCLUSION: This is the first comparative study between DNA methyltransferases expression profiles and global DNA methylation status in different phases of multiple myeloma patients. No significant correlation between the level of global methylation and the clinical stage of the unsorted cell population of patients with multiple myeloma was registered. Overexpression of the DNMT3A gene occurred in both sorted and unsorted cell populations of patients with multiple myeloma. This fact highlights the DNMT3A as a potential marker of multiple myeloma tumor progression. Moreover, we demonstrated comparable results in the expression of DNA methyltransferases in both sorted and unsorted cell populations. This is a promising result from the methodical point of view because when compared to samples of unsorted multiple myeloma cells, samples of sorted cells bring reduction of the number of possible analyses performed.

• MeSH
• DNA Methyltransferase 3A MeSH
• DNA MeSH
• Humans MeSH
• DNA Methylation * MeSH
• Multiple Myeloma * genetics   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

Gene inactivation of the cyclin-dependent kinase inhibitors p16INK4a, p15INK4b and p21WAF is frequently mediated by promoter gene methylation, whereas histone deacetylases (HDACs) control gene expression through their ability to deacetylate proteins. The effect of suberohydroxamic acid (SBHA) and 5-Aza-2'-deoxycytidine (Decitabine) (DAC) treatments on the transcription of CDKN2A, CDKN2B and CDKN1A genes, and their effects on molecular biological behavior were examined in two myeloma cell lines, RPMI8226 and U266, which differ in p53-functionality and IL-6 expression. In both tested myeloma cell lines, a non-methylated state of the CDKN2B gene promoter region was detected with normal gene expression, and the same level of p15INK4b protein was detected by immunocytochemical staining. Furthermore, in myeloma cells treated with SBHA and DAC alone, the expression of both p15INK4b and p21WAF was significantly upregulated in RPMI8226 cells (p53-functional, without IL-6 expression), whereas in the U266 cell line (p53 deleted, expressing IL-6) only p21WAF expression was significantly increased. Moreover, the analysis revealed that treatment with DAC induced DNMT3B enhancement in U266 cells. In conclusion, in myeloma cells with IL-6 expression, significantly increased DNMT3B expression indicated the tumorigenic consequences of 5-Aza-2'deoxycytidine treatment, which requires careful use in diseases involving epigenetic dysregulation, such as multiple myeloma (MM).

• MeSH
• Decitabine * pharmacology   MeSH
• DNA (Cytosine-5-)-Methyltransferases * genetics   metabolism   MeSH
• Epigenesis, Genetic * MeSH
• Cyclin-Dependent Kinase Inhibitor p15 genetics   metabolism   MeSH
• Cyclin-Dependent Kinase Inhibitor p16 genetics   metabolism   MeSH
• Interleukin-6 genetics   metabolism   MeSH
• Humans MeSH
• DNA Methylation MeSH
• Multiple Myeloma * genetics   metabolism   MeSH
• Cell Line, Tumor MeSH
• Tumor Suppressor Protein p53 genetics   metabolism   MeSH
• Cell Cycle Proteins genetics   metabolism   MeSH
• Gene Silencing MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

The immune system is important for elimination of residual leukemic cells during acute myeloid leukemia (AML) therapy. Anti-leukemia immune response can be inhibited by various mechanisms leading to immune evasion and disease relapse. Selected markers of immune escape were analyzed on AML cells from leukapheresis at diagnosis (N = 53). Hierarchical clustering of AML immunophenotypes yielded distinct genetic clusters. In the absence of DNMT3A mutation, NPM1 mutation was associated with decreased HLA expression and low levels of other markers (CLIP, PD-L1, TIM-3). Analysis of an independent cohort confirmed decreased levels of HLA transcripts in patients with NPM1 mutation. Samples with combined NPM1 and DNMT3A mutations had high CLIP surface amount suggesting reduced antigen presentation. TIM-3 transcript correlated not only with TIM-3 surface protein but also with CLIP and PD-L1. In our cohort, high levels of TIM-3/PD-L1/CLIP were associated with lower survival. Our results suggest that AML genotype is related to blast immunophenotype, and that high TIM-3 transcript levels in AML blasts could be a marker of immune escape. Cellular pathways regulating resistance to the immune system might contribute to the predicted response to standard therapy of patients in specific AML subgroups and should be targeted to improve AML treatment.

• MeSH
• Leukemia, Myeloid, Acute * diagnosis   genetics   MeSH
• B7-H1 Antigen genetics   MeSH
• Biomarkers MeSH
• Hepatitis A Virus Cellular Receptor 2 genetics   MeSH
• DNA Methyltransferase 3A * genetics   MeSH
• Humans MeSH
• Mutation MeSH
• Nucleophosmin * genetics   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

• MeSH
• Azacitidine analogs & derivatives   pharmacology   therapeutic use   MeSH
• DNA Modification Methylases genetics   metabolism   MeSH
• DNA (Cytosine-5-)-Methyltransferase 1 antagonists & inhibitors   MeSH
• Epigenesis, Genetic MeSH
• Hematopoietic Stem Cells MeSH
• Humans MeSH
• DNA Methylation * genetics   drug effects   MeSH
• Mutation genetics   MeSH
• Myelodysplastic Syndromes * drug therapy   genetics   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Review MeSH

Adiposis is reputed as a twin disease of type 2 diabetes and greatly harmful to human health. In order to understand the molecular mechanisms of adiposis, the changes of physiological, pathological, epigenetic and correlative gene expression were investigated during the adiposis development of C57BL/6J mice induced by long time (9 months) high-fat and high-sucrose diet (HFSD) sustainably. The results showed that mRNA transcription level of the Leptin, Glut4 and Glut2 genes have been obviously changed, which exhibit a negative correlation with methylation on their promoter DNA. The results also revealed that HFSD induced higher level of DNA methyltransferase 1 (DNMT1) in fat tissue might play important role in regulating the changes of methylation pattern on Glut4 and Leptin genes, and which might be one of the molecular mechanisms for the adiposis development.

• MeSH
• Adiposity physiology   MeSH
• Time Factors MeSH
• Diet, High-Fat adverse effects   trends   MeSH
• DNA (Cytosine-5-)-Methyltransferase 1 metabolism   MeSH
• Liver metabolism   pathology   MeSH
• Dietary Sucrose adverse effects   MeSH
• Leptin metabolism   MeSH
• Mice, Inbred C57BL MeSH
• Mice MeSH
• Glucose Transporter Type 4 metabolism   MeSH
• Adipose Tissue metabolism   pathology   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

• MeSH
• DNA (Cytosine-5-)-Methyltransferases metabolism   MeSH
• Epigenomics * MeSH
• Humans MeSH
• DNA Methylation MeSH
• MicroRNAs genetics   metabolism   MeSH
• Neoplasms genetics   pathology   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Editorial MeSH

Radiation exposure can evoke cellular stress responses. Emerging recognition that long non-coding RNAs (lncRNAs) act as regulators of gene expression has broadened the spectra of molecules controlling the genomic landscape upon alterations in environmental conditions. Knowledge of the mechanisms responding to low dose irradiation (LDR) exposure is very limited yet most likely involve subtle ancillary molecular pathways other than those protecting the cell from direct cellular damage. The discovery that transcription of the lncRNA PARTICLE (promoter of MAT2A- antisense radiation-induced circulating lncRNA; PARTICL) becomes dramatically instigated within a day after LDR exposure introduced a new gene regulator onto the biological landscape. PARTICLE affords an RNA binding platform for genomic silencers such as DNA methyltransferase 1 and histone tri-methyltransferases to reign in the expression of tumor suppressors such as its neighboring MAT2A in cis as well as WWOX in trans. In silico evidence offers scope to speculate that PARTICLE exploits the abundance of Hoogsten bonds that exist throughout mammalian genomes for triplex formation, presumably a vital feature within this RNA silencer. PARTICLE may provide a buffering riboswitch platform for S-adenosylmethionine. The correlation of PARTICLE triplex formation sites within tumor suppressor genes and their abundance throughout the genome at cancer-related hotspots offers an insight into potential avenues worth exploring in future therapeutic endeavors.

• MeSH
• Radiation Dosage MeSH
• DNA (Cytosine-5-)-Methyltransferase 1 genetics   MeSH
• Genome, Human radiation effects   MeSH
• Genomics MeSH
• Histone Methyltransferases genetics   MeSH
• Humans MeSH
• Methionine Adenosyltransferase genetics   MeSH
• Tumor Suppressor Proteins genetics   MeSH
• Neoplasms genetics   radiotherapy   MeSH
• WW Domain-Containing Oxidoreductase genetics   MeSH
• Promoter Regions, Genetic genetics   MeSH
• Radiation Exposure adverse effects   MeSH
• Gene Expression Regulation, Neoplastic radiation effects   MeSH
• RNA, Long Noncoding genetics   MeSH
• RNA Interference radiation effects   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

The fusion of sperm and oocytes determines the fertilization competence and subsequent development of embryos, which, in turn, can be affected by various proteins and DNA methylation. However, several factors in this whole regulation process remain unknown, especially in yaks. Here, we report that fibroblast growth factor 10 (FGF10) is an important growth factor that can enhance the maturation rate of yak oocytes and the motility of frozen spermatozoa. Subsequent blastocyst quality was also improved by increasing the total cell number and level of pregnancy-associated protein in blastocysts. These effects were significantly high in the group that received the 5 ng/ml FGF10 treatment, during both in vitro maturation (IVM) and capacitation. Our data show that the effects of FGF10 were dose-dependent at vital steps of embryogenesis in vitro. Furthermore, quantitative polymerase chain reaction, western blot analysis, and immunofluorescence demonstrated that the levels of CD9, CD81, DNMT1, and DNMT3B in both mature cumulus-oocyte complexes and capacitated sperms were regulated by FGF10, which was also highly expressed in the group treated with 5 ng/ml FGF10 during both IVM and capacitation. From our present study, we concluded that FGF10 promotes yak oocyte fertilization competence and subsequent blastocyst quality, and could also regulate CD9, CD81, DNMT1, and DNMT3B to optimize sperm-oocyte interactions and DNA methylation during fertilization.

• MeSH
• Tetraspanin 28 genetics   metabolism   MeSH
• Tetraspanin 29 genetics   metabolism   MeSH
• Blastocyst drug effects   physiology   MeSH
• DNA (Cytosine-5-)-Methyltransferases genetics   metabolism   MeSH
• DNA (Cytosine-5-)-Methyltransferase 1 genetics   metabolism   MeSH
• Embryonic Development drug effects   genetics   physiology   MeSH
• Fertilization in Vitro veterinary   MeSH
• Fertilization drug effects   genetics   physiology   MeSH
• Fibroblast Growth Factor 10 administration & dosage   physiology   MeSH
• In Vitro Oocyte Maturation Techniques veterinary   MeSH
• RNA, Messenger genetics   metabolism   MeSH
• Oocytes drug effects   physiology   MeSH
• Cattle embryology   genetics   physiology   MeSH
• Pregnancy MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Cattle embryology   genetics   physiology   MeSH
• Pregnancy MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH