Project description:RA signalling regulated endothelial to hematopoietic transition and HSC generation. EB- or FL-derived HSPC were profiled before (d0) or after (d6) 6 days of treatment with 0.2uM AM580 on OP9, and after 6 additional days of expandion of OP9 (d12) without treatment.
Project description:HOXA7 regulates FL-HSPC self-renewal in vitro and in vivo. We profiled EB-HSPCs after HOXA7 overexpression (EB-HOXA7), or with a control vector (EB-CTR), to assess the gene expression programs regulated by HOXA7. CD34+CD38-CD43+CD90+ HSPCs were infected with lentiviral FUGW vector either empty (FUGW-GFP) or encoding HOXA7(FUGW-GFP-HOXA7) protein. Cells were expanded on op9 for 15 days and than sorted for GFP HSPC immunophenotype.
Project description:HOXA7 regulates FL-HSPC self-renewal in vitro and in vivo. We profiled EB-HSPCs after HOXA7 overexpression (EB-HOXA7), or with a control vector (EB-CTR), to assess the gene expression programs regulated by HOXA7.
Project description:ATAC-seq and RNA-seq were used to compare the chromatin states and corresponding transcriptomes of developing HSPC. We found that the chromatin states and transcriptomes change markedly when HSPCs migrate from PL to FL and from FL to BM. Then we compared chromatin among E12.5 PL, E12.5 FL, and E16.5 FL, the proportion of fragments in the nucleosome-free region significantly increases in E12.5 and E16.5 FL-HSPCs compared with E12.5 PL-HSPCs; the motifs of ETS and Runt are enriched at the opeing or closing peaks from E12.5 PL to E12.5 FL and E12.5 FL to E16.5 FL, and the expression of these factors is either up-regulated or down-regulated; the motifs of bZIP and Zf are enriched at the closing and opening peaks from E12.5 PL to E12.5 FL and E12.5 FL to E16.5 FL respectively, In line, the expression of these factors is down-regulated or up-regulated.
Project description:ATAC-seq and RNA-seq were used to compare the chromatin states and corresponding transcriptomes of developing HSPC. We found that the chromatin states and transcriptomes change markedly when HSPCs migrate from PL to FL and from FL to BM. Then we compared chromatin among E12.5 PL, E12.5 FL, and E16.5 FL, the proportion of fragments in the nucleosome-free region significantly increases in E12.5 and E16.5 FL-HSPCs compared with E12.5 PL-HSPCs; the motifs of ETS and Runt are enriched at the opeing or closing peaks from E12.5 PL to E12.5 FL and E12.5 FL to E16.5 FL, and the expression of these factors is either up-regulated or down-regulated; the motifs of bZIP and Zf are enriched at the closing and opening peaks from E12.5 PL to E12.5 FL and E12.5 FL to E16.5 FL respectively, In line, the expression of these factors is down-regulated or up-regulated.
Project description:HOXA7 regulates FL-HSPC self-renewal in vitro and in vivo. We profiled FL-HSPCs after HOXA7 knockdown, to assess the gene expression programs regulated by HOXA7.
Project description:The aim of this study was to investigate the effect of T0901317 LXR agonist on Retinoic Acid Receptor (RAR) family gene expression. T0901317 alone increase RARalpha gene expression in monocytes and the effects of the co treatment of the cells with T0901317 and a specific RARalpha agonist (AM580) were further studied. For this monocytes were treated for 24h with DMSO or T0901317 to induce RARalpha expression and in a second step cells were treated or not with AM580 for another 24h period. Gene expression was analysed for all the treatments (T0901317, AM580 or T0901317+AM580) and we focussed on genes which expression was synergitically induced by the co treatment as compared to the cells treated either by T0901317 or AM580.
Project description:Osteolineage cell-derived extracellular vesicles (EVs) play a regulatory role in hematopoiesis and have been shown to promote the ex vivo expansion of human hematopoietic stem and progenitor cells (HSPCs). Here, we demonstrate that EVs from different human osteolineage sources do not have the same HSPC expansion promoting potential. Comparison of stimulatory and non-stimulatory osteolineage EVs by next-generation sequencing and mass spectrometry analyses revealed distinct microRNA and protein signatures identifying EV-derived candidate regulators of ex vivo HSPC expansion. Accordingly, the treatment of umbilical cord blood-derived CD34+ HSPCs with stimulatory EVs altered HSPC transcriptome, including genes with known roles in cell proliferation. An integrative bioinformatics approach, which connects the HSPC gene expression data with the candidate cargo in stimulatory EVs, delineated the potentially targeted biological functions and pathways during hematopoietic cell expansion and development. In conclusion, our study gives novel insights into the complex biological role of EVs in osteolineage cell-HSPC crosstalk and promotes the utility of EVs and their cargo as therapeutic agents in regenerative medicine.