Project description:Sox17-Erg direct reprogramming converts neonatal murine cardiac fibroblasts into induced endothelial cells. This data evaluates the conversion over time of the Sox17-Erg iECs compared to control condition.
Project description:Direct conversion of somatic cells into neurons holds great promise for regenerative medicine. However, neuronal conversion is relatively inefficient in human cells compared to mouse cells. It has been unclear what might be the key barriers to reprogramming in human cells. We recently elucidated an RNA program mediated by the polypyrimidine tract binding protein PTB to convert mouse embryonic fibroblasts (MEFs) into functional neurons. In human adult fibroblasts (HAFs), however, we unexpectedly found that invoking the documented PTB–REST–miR-124 loop generates only immature neurons. We now report that the functionality requires sequential inactivation of PTB and the PTB paralog nPTB in HAFs. Inactivation of nPTB triggers another self-enforcing loop essential for neuronal maturation, which comprises nPTB, the transcription factor BRN2, and miR-9. These findings suggest that two separate gatekeepers control neuronal conversion and maturation and consecutively overcoming these gatekeepers enables deterministic reprogramming of HAFs into functional neurons.
Project description:Direct cell conversion is now expected to apply to therapeutic purposes. Although that has been succeeded in several cell types, the mechanism or general way to identify the key transcription factors are still unclear. In addition, most of the cases are not completely identical with the target cells. In previous work, we suggested that cell status is maintained by a homeostatic network of limited number of TFs and no single transcription factor is both necessary and sufficient to drive the differentiation process. Here, identifying the key TFs of human monocyte by combining comparative gene expression analysis and literature based text-mining, we mimicked the monocytic regulatory network in human dermal fibroblasts to induce direct cell conversion of the fibroblasts to monocytes. We suggested that although it is a primary master TF, single TF is not sufficient to induce the direct cell conversion and orchestrated TF regulation is necessary to complete the cell conversion. Total RNA obtained from human dermal fibroblasts(FIB), human CD14+ monocytes(MON), mock lentivirus vector transduced fibroblasts (FIB-mock), SPI1 transduced fibroblasts (FIB-SPI1), and SPI1, CEBPA, MNDA, IRF8 transduced fibroblasts(FIB-4Fs). The fold change was computed compared with fibroblasts or FIB-mock.
