Although very preliminary, these data suggest that different HGPS vascular cells could autonomously upregulate or induce the expression of bone-related genes
Although very preliminary, these data suggest that different HGPS vascular cells could autonomously upregulate or induce the expression of bone-related genes. In addition to (iSMCs) or (iVECs). of fibroblasts, SMCs and endothelial cells employed in the study. elife-54383-supp1.docx (11K) GUID:?246E13A8-49D2-4BCD-8B48-938D7DCEC496 Supplementary file 2: Details on DE genes between young vs?old iVECs, young vs old iSMCs, normal vs HGPS iSMCs. Details on clustering analyses comparing iVECs, primary VECs and fibroblasts as well as iSMCs, primary SMCs and fibroblasts. elife-54383-supp2.docx (9.0K) GUID:?31AD9787-33A2-43CE-B5B1-9BBFAFDDF65D Supplementary file 3: Details on clustering analyses comparing induced vs primary cells. This dataset refers to data presented in Figure 1G and I. elife-54383-supp3.docx (8.7K) GUID:?AFA8F063-518B-402B-81D9-6262CA6077DB Supplementary file 4: Code used for data analyses. elife-54383-supp4.docx (8.6K) GUID:?AC97A910-6D28-416F-952D-48A3374BA1BA Supplementary file 5: Cell identity gene analysis in reprogrammed cells. The table collects relevant information on key cell identity genes expressed by reprogrammed and Ivacaftor hydrate primary cells. elife-54383-supp5.docx (9.2K) Ivacaftor hydrate GUID:?2A11D0E7-2B95-4EAF-92A5-BB5A3CBF64F1 Supplementary file 6: Cell identity gene analysis in young vs old reprogrammed cells. The table collects relevant information on key cell identity genes expressed by reprogrammed cells derived from young and old donors. elife-54383-supp6.docx (8.6K) GUID:?A5750197-CA5C-46CC-9319-18C41F742EE5 Supplementary file 7: The table collects relevant information on the source Rabbit Polyclonal to OR8J3 of human serum employed in the study. elife-54383-supp7.docx (8.5K) GUID:?5C34C7F2-8814-46C8-AA6A-525D859CA045 Transparent reporting form. elife-54383-transrepform.docx (250K) GUID:?20E6A76F-A757-43AF-8440-98B7CC6D8D02 Data Availability StatementSequencing data have been deposited in GEO under accession code “type”:”entrez-geo”,”attrs”:”text”:”GSE140898″,”term_id”:”140898″GSE140898. The following dataset was generated: Bersini S, Schulte R, Huang L, Tsai H, Hetzer MW. 2019. Direct reprogramming of fibroblasts identifies signatures of vascular Ivacaftor hydrate dysfunction in physiological aging and Hutchinson-Gilford Progeria Syndrome. NCBI Gene Expression Omnibus. GSE140898 The following previously published dataset was used: Quertermous T. 2018. Coronary artery disease genes SMAD3 and TCF21 promote opposing interactive genetic programs that regulate smooth muscle cell differentiation and disease risk. NCBI Gene Expression Omnibus. GSM3175518 Abstract Vascular dysfunctions are a common feature of multiple age-related diseases. However, modeling healthy and pathological aging of the human vasculature represents an unresolved experimental challenge. Here, we generated induced vascular endothelial cells (iVECs) and smooth muscle cells (iSMCs) by direct reprogramming of healthy human fibroblasts from donors of different ages and Hutchinson-Gilford Progeria Syndrome (HGPS) patients. iVECs induced from old donors revealed upregulation of and KD VECs demonstrated a recovery in vascular permeability. We found that iSMCs from HGPS donors overexpressed bone morphogenetic protein (alone, which is a master regulator of VEC development and early vasculogenesis (Qiu and Hirschi, 2019), was sufficient to in vitro reprogram human skin fibroblasts into functional VECs (Morita et al., 2015). Similarly, the induced expression of myocardin (or (Figure 1G, cluster 2) and multiple endothelial-specific genes (e.g. and compared to primary endothelial cells (Figure 1I, cluster 3). However, the expression level of (an important gene defining endothelial cell identity) was 6.62 log2FoldChange (FC) higher in primary VECs vs. fibroblasts and 6.38 log2FC higher in iVECs vs. fibroblasts. At the same time, another endothelial-specific gene (i.e. and compared to fibroblasts (Supplementary file 5). Notably, the expression of was lower in reprogrammed vs. primary SMCs. Furthermore, key skin fibroblast cell identity genes (e.g. and and was comparable in reprogrammed cells derived from young and old donors (log2FC?=?0.03 for and log2FC?=?0.19 for comparing young and old cells; no statistical difference comparing young and old cells). DE analysis highlighted a set of genes (21 DE genes for iSMCs; 9 DE genes for iVECs) that changed according to the age of the donor for either iSMCs (Figure 2A) or iVECs (Figure 2B). We then analyzed the expression of a few cell identity genes in iSMCs or iVECs derived from young and old donors without finding obvious age-related differences (Supplementary file 6 and Figure 2figure supplement 1). Together with the similar expression of MYOCD and ETV2 in cells derived from young and old donors, these data suggest that the reprogramming efficiency should not be affected by the age of the donor and that potential differences between young and old cells do not seem to be due to different levels of reprogramming. Open in a separate window Figure 2. Vascular cells reprogrammed from young vs. old donors show gene expression and functional differences.Heat-map representing DE genes between reprogrammed vascular cells from young (N?=?3) vs. old (N?=?3) donors (iSMCs (A), iVECs (B)). Z score?=??1.5. DE genes with log2FC?>?1 and FDR?0.05. Quantification of BMP2.