Project description:Haploid embryos can be induced from cultured immature pollen following stress treatment. In Brassica napus, the application of the histone/lysine deacetylase (HDAC/KDAC) inhibitor trichostatin A (TSA) to pollen cultures enhances the production of differentiated embryos and embryogenic callus when applied together with heat stress (Li et al., 2014). To identify genes associated with the induction of haploid embryogenesis and to investigate which genes may be responsible for TSA-induced lipid and starch accumulation in the embryogenic structures, we compared the transcriptomes of pollen cultures treated with heat stress and 0.05 µM TSA to those of untreated pollen cultures, both at two days.
Project description:Haploid embryos can be induced from cultured immature pollen following a stress treatment. In Brassica napus, application of the histone/lysine deacetylase (HDAC/KDAC) inhibitor trichostatin A (TSA) to pollen cultures enhances the production of differentiated embryos and embryogenic callus when applied together with heat stress (Li et al., 2014). To identify genes associated with the induction of B. napus haploid embryogenesis, we compared the transcriptomes of untreated pollen cultures and pollen cultures treated with either heat-stress or heat-stress plus TSA.
Project description:Western Blot showed that epigenetic control of the mouth dimorphism in P. pacificus correlates with histone 4 acetylation. This result was validated by mass spectrometry. This experiment profiled histone acetylation in the presence and absence of the histone deacetylase (HDAC) inhibitor Trichostatin A (TSA). Histone acetylation was also examined in the presence of another HDAC inhibitor (Na-butyrate), which served as an additional negative control because unlike TSA, it did not induce a phenotypic change.
Project description:Treatment of the acute T cell leukemia cell line Jurkat in vitro with the most potent histone deacetylase inhibitor, Trichostatin A induced apoptosis in a dose-dependent manner. Analysis of the drug-treated Jurkat cells using high throughput genome-wide gene expression profiling indicated the upregulation as well as downregulation of several genes crucial for cellular functions. Use of quantitative reverse transcriptase-mediated polymerase chain reaction validated the regulation of selected genes in drug-treated cells. Collectively, this study has unraveled the genes involved in epigenetic regulation of the T cell leukemia. Microarray analysis was performed to determine the changes in global gene expression profiles during apoptosis of a T cell leukemia induced by the histone deacetylase inhibitor, Trichostatin A. Drug treatment induced the upregulation and repression of a number of genes.
Project description:Bovine cloned and IVF blastocysts at day 7 were collected and used for comparative transcriptomics. A potent histone deacetylase inhibitor, Trichostatin A (TSA), was used to understand the effect of assisted epigenetic modification on the transcriptional profile of SCNT blastocysts and to identify specific genes or categories of genes affected in the developing embryos.
Project description:The haploid multicellular male gametophyte of plants, the pollen grain, is a terminally differentiated structure whose function ends at fertilization. Unlike pollen grains, the immature gametophyte retains its capacity for totipotent growth when cultured in vitro. Haploid embryo production from cultured immature male gametophytes is a widely used plant breeding and propagation technique that was described nearly 50 years ago, but one that is poorly understood at the mechanistic level. Using a chemical approach, we show that the switch to haploid embryogenesis is controlled by the activity of histone deacetylases (HDACs). Blocking HDAC activity with trichostatin A (TSA) in cultured immature male gametophytes of Brassica napus leads to a large increase in the proportion of cells that switch from pollen to embryogenic growth. Embryogenic growth is enhanced by, but not dependent on, the high temperature stress that is normally used to induce haploid embryogenesis in B. napus. The immature male gametophyte of Arabidopsis thaliana, which is recalcitrant for haploid embryo development in culture, also forms embryogenic cell clusters after TSA treatment. TSA treatment of immature male gametophytes for as little as eight hours was accompanied by hyperacetylation of histones H3 and H4, and by the upregulation of genes involved in cell-cycle progression, the auxin pathway and cell wall catabolism pathways. We propose that the totipotency of the immature male gametophyte in planta is kept in check by an HDAC-dependent mechanism, and that high temperature or other stresses used to induce haploid embryo development in culture impinge on this HDAC-dependent pathway. 8 samples were analyzed. We generated the following pairwise comparisons between treatment and the corresponding mock treatment: TSA+CHX (2 replicates) vs CHX (2 replicates); TSA (2 replicates) vs DMSO (2 replicates).
Project description:Somatic cell nuclear transfer (SCNT) enables gaining of totipotency by reprogramming nuclei of terminally differentiated donor cell. Recent studies have clearly demonstrated that intervention of epigenetic networks can significantly elevate both in vitro and in vivo development potential of NT embryos. Specifically, trichostatin A (TSA), a kind of histone deacetylase inhibitors (HDACi), was proved to functionally works during cloning in various mammal systems. However, how it modulates histone acylation lacks careful illustration. Here, we systematically evaluate the effect and limitation of TSA during SCNT embryo development by generating genome-wide H3K9ac maps. In addition, a Dux-dependent 2-cell (2C) activation deficiency is observed in SCNT embryos as compared with their natural fertilized counterparts. Strikingly, a refined Dux supplement can successfully assist SCNT embryos in overcoming the 2C activation defect and further promotes the overall cloning efficiency. Together, our study for the first time reveals the regulation mechanism of histone marker H3K9ac in SCNT and provides the new insight of Dux during embryogenesis.
Project description:The aim of this study is to determine the efficacy of combining the histone deacetylase (HDAC) inhibitor sodium valproate (VPA) with anti-EGFR monoclonal antibody (panitumumab or cetuximab) maintenance in the first-line treatment of patients with RAS wild type metastatic CRC.
Project description:This study is an open label non randomized study of hydroxychloroquine (HCQ) with histone deacetylase (HDAC) inhibitor Vorinostat in patients with advanced solid tumors to determine the maximum tolerated dose (MTD) and to evaluate the safety and antitumor activity of this drug combination.
Project description:Neurosphere cultures were established from individually isolated E 14.5 forebrains of mouse embryos that carry a bcl2 transgene. Single-cell suspensions were prepared, seeded at 1x105 cells/ml and treated for two days with 150 nM Trichostatin A (TSA; histone deacetylase inhibitor), 500 nM 5-Aza-2-deoxycytidine (AzaC; demethylating agent) or both compounds, or left untreated. Two independent experiments were performed.