co-culture system mimicking early-stage atherosclerosis. Preliminary drug screening with FDA-approved anti-inflammatory drugs, rosuvastatin, and reparixin has shown promising potential for high-throughput applications (Liu et al, unpublished). This innovative approach holds promise for accelerating drug discovery and advancing therapeutic interventions for atherosclerosis in WS patients. Experimental MethodsRecently, we established disease-specific iPSCs from patients with WS and repaired the genetic mutation in one of the alleles of the WRN gene locus, and established gene-corrected (gc) WS-iPS cells 6). By utilizing those iPS cells, we differentiated macrophages (iMφs), vascular endothelial cells (iVECs), and vascular smooth muscle cells (iVSMCs). Following this, we developed a 2D in vitro model of atherosclerosis using these three cell types derived from WS patient-specific iPS cells with a uniform genetic background and systematically compared them with corresponding cells derived from healthy donors 5). Furthermore, we immortalized the cells with doxycycline (Dox)-inducible overexpressing lentiviral vectors harboring c-MYC, BMI1, and Bcl-XL (Paul et al, unpublished) (Japanese Patent application: 2021-109513). Results and DiscussionCompared to healthy controls, we observed no abnormalities in WS-iPS-derived VECs (WS-iVECs) and WS-iPS-derived iVSMCs (WS-iVSMCs). However, WS-iPS-derived Mφs (WS-iMφs) showed reduced proliferation, increased senescence, and apoptosis, which were rescued in gcWS-iMφs (Fig. 1A). In a co-culture experiment, WS-iMφs induced endothelial dysfunction and promoted a synthetic phenotype in WS-iVSMCs (Fig. 1B and C). RNA-seq and ATAC-seq revealed upregulated type I IFN signaling and reduced chromatin accessibility at key transcription factor binding sites. Notably, IRF, CEBP, and JUN-AP-1 motifs were enriched in WS-iMφs, whereas GATA, E2F7, TCF, and PBX1 were enriched in healthy-iMφs. These changes contributed to impaired cellular homeostasis, increased inflammation, and the pathological features observed in WS-iMφs compared to healthy and gene-corrected iMφs 5).Figure 1. Effects of inflammatory iMϕs on iPSC-derived vascular cells― 245 ―
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