Journal: bioRxiv
Article Title: Braf -mutant Schwann cells divert to a repair phenotype to induce congenital demyelinating neuropathy
doi: 10.1101/2024.04.24.590951
Figure Lengend Snippet: (A) Schematic representation of the differentiation of human induced pluripotent stem cells (hiPSCs) into SC, derived from two WT (AG08H, HFF15) or two CFC patients carrying the BRAF p.Q257R mutation (RMK0056C, RMK0138C). D0: hiPSCs; D6: multipotent neural crest-like cells (NCCs); D12: presumably committed NCC; D24: SC precursors (SCP); D31: immature SC (iSCs); D90: myelin-competent SC. Representative bright-field images of cells during the differentiation protocol are shown at D13, D19, a day after passage on D32 and at D95-100, after five days’ exposure to ascorbic acid. Scale bar = 100 µm for all frames. (B) Euler diagram showing the number of common/specific dysregulated genes (p-value < 0.05, Log2FC > 1 or < -1) in RMK0056C and RMK0138C mutant cells compared to control cells (HFF15) at D21 of differentiation. (C) Bubble plot of the enrichment analysis performed on the common 380 dysregulated genes in RMK0056C and RMK0138C. Gene set names from the REACTOME database are indicated on the y-axis. Gene sets are ranked according to significance level (-log adjusted p-value). The size of the dots represents the number of queried genes present in the corresponding pathway, and the color corresponds to the gene ratio (number of queried genes present in the gene set / gene set size). (D) Enrichment plot for Egr2 and Sox10 mediated initiation of SC myelination using Reactome database, corresponding to the significant gene set downregulated in RMK0056C and RMK0138C cell lines after 30 days of differentiation. (E) Enrichr-KG analysis of the most significantly enriched terms across three additional databases (Descartes Cell Types and Tissue 2021, Jensen DISEASES and CCLE Proteomics 2020) highlight the close lineage relationship between Schwann cell progenitors and either melanocytes (“melanoma” and “skin cancer”, the A2058 cell line) or neuroendocrine cells (“visceral neurons in lung/heart”, the SKNAS and NCIH446 cell lines).
Article Snippet: The AG08H and hFF15 clones of healthy donor hiPSC lines were derived (CytoTune IPS 2.0 reprograming kit, Thermofisher) from commercial cell lines consented for research, AG08498 (RRID:CVCL_1Y51) and HFF-1 SCRC-1041 (RRID:CVCL_3285) respectively, as described ( ).
Techniques: Derivative Assay, Mutagenesis, Control