Project description:Introduction Current research efforts attempt to increase therapy efficacy in diffuse large B-cell lymphoma (DLBCL). Here, we evaluated the effects of small molecule inhibitor Ro 08-2750 on therapy-induced cytotoxicity in DLBCL. Material and methods First, we established Ro 08-2750 treatment in DLBCL by verifying cell toxicity via cell viability and colony formation assays. In a next step, functional effects reducing cell viability, e.g. cell cycle dysregulation and apoptosis, were analyzed via flow cytometry. Then, we evaluated effects of Ro 08-2750 on cytotoxic therapy in cell viability and colony formation assays. After confirming an increase in irradiation-induced DNA damage via γH2AX assay, an immunoblot-based assay was used to detect changes in DNA damage response. Lastly, we analyzed Ro 08-2750-induced changes in gene and protein expression via RT-qPCR, RNA sequencing, western blot and proteomic analysis. Results Ro 08-2750 reduces cell viability dose-dependently in DLBCL. Functionally, the decrease in cell viability is combined with a cell cycle dysregulation, specifically an increase in S phase with reduced G2/M phase, and an increase in apoptosis. DNA synthesis is reduced during S phase. Assessing the therapeutical relevance, Ro 08-2750 sensitized DLBCL cell lines to irradiation but not chemotherapy and increased irradiation-induced double strand breaks. We found a dysregulation in CHK2 phosphorylation. Moreover, Ro 08-2750 dysregulated DNA damage response in genomic and proteomic analysis. Conclusion We identified Ro 08-2750 as a potential radiosensitizer in DLBCL. Therefore, our results indicate the potential therapeutical benefit of targeting RNA binding proteins
Project description:Dual targeting of PRMT5 and MSI2 inhibited synergisticly B-cells proliferation. Thus, next-generation sequencing was performed to evaluate the effect of the inhibitors of PRMT5 and MSI2, GSK-591 and Ro, respectively, in the global transcriptome of the mantle cell lymphoma cell line, Z-138.
Project description:Whole genome sequencing was performed on Halomonas sp. DSM7328 both original strain and evolved population in low dissolved oxygen (DO) level at 0.5% DO. The evolved Halomonas sp. DSM7328 was analysed and compared to the original strain.
Project description:Puccinia graminis f. sp. tritici is the cause of wheat stem rust. A microarray was designed from genes predicted from the P. graminis f. sp. tritici genome assembly, and gene expression measured for four conditions which include wheat or barley infecting growth stages initiated by urediniospores. mRNA was prepared from fresh urediniospores, uredinospores germinated for 24 hr, wheat seedlings infected with urediniospores for 8 days, and barley seedlings infected with urediniospores for 8 days. The asexual uredinial infection cycle on wheat produces additional urediniospores, which can start new cycles of wheat infection and are readily spread by aerial transport. This expression data is further described in Duplessis et al, Obligate Biotrophy Features Unraveled by the Genomic Analysis of the Rust Fungi, Melampsora larici-populina and Puccinia graminis f. sp. tritici
Project description:Corneal epithelial stem cells reside in the limbus that is the transitional zone between the cornea and conjunctiva, and are essential to maintain the homeostasis of corneal epithelium. However, their characterization is poorly understood. Therefore, we constructed gene expression profiles of limbal epithelial SP and non-SP cell using RNA-sequencing. As a result, limbal epithelial SP cells have immature cell phenotypes with endothelial/mesenchymal cell markers, while limbal epithelial non-SP cells have epithelial progenitor cell markers.