Project description:To investigate class A G protein-coupled receptors (GPCR)-targeted drugs in the regulation of osteogenic differentiation, we investigated the effects of drugs using mesenchymal stromal cells. By applying microarray dataset, we idntified the mRNA expressions profiles in hDPSCs.
Project description:To investigate class A G protein-coupled receptors (GPCR)-targeted drugs in the regulation of osteogenic differentiation, we investigated the effects of drugs using mesenchymal stromal cells.
Project description:To investigate class A G protein-coupled receptors (GPCR)-targeted drugs in the regulation of osteogenic differentiation, we investigated the effects of drugs using mesenchymal stromal cells. We conducted whole-transcriptome analysis using bulk RNA sequencing (RNA-seq) of six drugs.
Project description:MDM2 inhibitor remarkably induced biomineralization in hDPSCs but it remained the problem that p53 activation is insufficient due to MDM2-p53 autoregulatory feedback loop. To overcome the limitation of the MDM2 inhibitors, we applied proteolysis targeting chimera (PROTAC), a technology that degrades a protein of interest (POI) by intracellular ubiquitin-proteasome system. Hence, we propose a strategy to induce hard tissue regeneration by MDM2-targeting PROTAC technology. We selected Nutlin-3 of POI ligand among various MDM2 inhibitors based on the screening process, and the selected CRBN of E3 ligase ligand. The MDM2-PROTAC synthesis platform was designed by each ligand combination. By performing the degradation test for selected compounds, we evaluated the characteristics of the developed MDM2 PROTAC such as the maximal degradation concentration (DCmax), the half of maximal degradation concentration (DC50), and half-lifetime. To investigate gene expression profiling of MDM2-targeting small molecules, we conducted RNA-sequencing under MDM2 PROTAC and Nutlin-3 (POI ligand) treated conditions. We not only confirmed a robust effect on biomineralization by MDM2-targeting small molecules but also demonstrated the potent osteogenic differentiation ability of MDM2 PROTAC when compared to an MDM2 inhibitor. MDM2-PROTAC significantly increased mRNA levels of osteogenic differentiation marker genes. Also, the significant bone generation effect of MDM2 PROTAC was validated in an ovariectomy (OVX)-induced osteoporosis animal model. Through these results, it is expected that a new therapeutic modality for hard tissue regeneration will be possible, and the application range of the PROTAC system can be expanded.
Project description:The growth behavior of plant roots on tilted, hard agar surfaces is determined by many basic cellular processes, including microtubule dynamics and cell wall expansion. Among Arabidopsis thaliana accessions there is natural variation for these behaviors, including one known as skewing or slanting. The root skewing pattern on hard, tilted agar surfaces may be a clue to adaptations of an accession to its environment. Here, we compare expression profiles of two accessions with diverse skewing behavior grown on the wave assay, which consists of seedlings growing two days vertically and 3 days tilted on hard agar plates. Cvi has a strong skew on tilted, hard agar sufaces, and Ler-2 has a weaker one. We also include a near isogenic line, 170G-55-16 a.k.a HGI2.1, that is mostly Ler-2 in background but has a segment of Cvi introgressed into chromosome 2. This line has an intermediate skew between its two parents.
Project description:The growth behavior of plant roots on tilted, hard agar surfaces is determined by many basic cellular processes, including microtubule dynamics and cell wall expansion. Among Arabidopsis thaliana accessions there is natural variation for these behaviors, including one known as skewing or slanting. The root skewing pattern on hard, tilted agar surfaces may be a clue to adaptations of an accession to its environment. Here, we compare expression profiles of two accessions with diverse skewing behavior grown on the wave assay, which consists of seedlings growing two days vertically and 3 days tilted on hard agar plates. Cvi has a strong skew on tilted, hard agar sufaces, and Ler-2 has a weaker one. We also include a near isogenic line, 170G-55-16 a.k.a HGI2.1, that is mostly Ler-2 in background but has a segment of Cvi introgressed into chromosome 2. This line has an intermediate skew between its two parents. 3 biological replicates of each of 3 genotypes (Cvi, Ler-2, and 170G-55-16/HGI2.1) were subjected to the wave assay. After the assay, approximately 600 root tips from each biological replicate were pooled for RNA extraction and hybridization on the Affymetrix ATH1 microarray.