Project description:Four healthy iPSC lines were differentiated into ECs. iPSC-ECs were then treated with and without e-cigarette extraction for 24 hours.
Project description:Two healthy iPSC lines (78 and 273) were differentiated into ECs. iPSC-ECs were then treated with e-cig liquids RY4 (RY) or Marcado (MAR) with 18mg/ml of nicotine (18) and without nicotine (0).
Project description:Two healthy iPSC lines were differentiated into ECs. iPSC-ECs were then treated under normoglycemic (5.5mM glucose) or hyperglycemic (33mM glucose) conditions with and without MDL-28170 (2.5 uM) for 72 hours.
Project description:Here we report genes upregulated for iPSC-ECs cultured in PEG hydrogels relative to control cells on tissue culture polystyrene (TCP) surfaces included adhesion, matrix remodeling, and Notch signaling pathway genes relevant to in vivo vascular development.
Project description:Discovery of small molecules that correct gene networks dysregulated in human disease may allow identification of therapies that treat disease at its fundamental basis by leveraging mechanism-based data. Here, we report the first broad gene network-based drug screen, which led to discovery of a drug candidate that effectively treats aortic valve disease in an animal model. We previously reported haploinsufficiency of NOTCH1 (N1) as a genetic cause of human aortic valve thickening and calcification, the third most common form of heart disease, and described the resulting gene network dysregulation in human induced pluripotent stem cell (iPSC)-derived endothelial cells (ECs). We exposed isogenic N1+/+ or N1+/– iPSC-derived ECs to each of 1595 small molecules or control and developed a machine learning approach that accurately distinguished WT or N1-haploinsufficient cells based on expression of 119 genes assayed by targeted RNA-sequencing. 9 small molecules corrected the gene network of N1+/– ECs sufficiently to be classified as WT. Among hits tested in vivo, the estrogen receptor-related alpha inverse agonist XCT790 significantly reduced aortic valve thickening, calcification, and stenosis in N1-haploinsufficient mice with shortened telomeres, which model the range of age-dependent cardiac disease observed in humans. This strategy, made feasible by human iPSC technology, next generation sequencing approaches, and machine learning, may represent a more effective path for drug discovery compared to conventional screening approaches.
Project description:While microRNAs have extensively been investigated in cancer research, little is known regarding their response to noxious agents in apparently healthy tissues. We analyzed the expression of 484 miRNAs in the lung of rats exposed to environmental cigarette smoke (ECS) for 28 days. ECS downregulated 126 miRNAs (26.0%) at least two-fold and 24 miRNAs more than three-fold. We previously demonstrated that 107 of 4858 genes (2.9%) and 50 of 518 proteins (9.7%) were upregulated by ECS in the same tissue, consistently with the role of microRNAs as negative regulators of gene expression. The most remarkably downregulated microRNAs belonged to the families of let-7, miR-10, miR-26, miR-30, miR-34, miR-99, miR- 122, miR-123, miR-124, miR-125, miR-140, miR-145, miR-146, miR-191, miR-192, miR-219, miR-222, and miR-223, which regulate stress response, apoptosis, proliferation, angiogenesis, and expression of genes. In contrast, miR-294, an inhibitor of transcriptional repressor genes, was upregulated by ECS. There was a strong parallelism in dysregulation of rodent microRNAs and their human homologues, which are often transcribed from genes localized in fragile sites deleted in lung cancer. Five ECS-downregulated microRNAs are known to be affected by single nucleotide polymorphisms. Thus, changes in microRNA expression are an early event following exposure to cigarette smoke. Keywords: microRNAs ⢠gene expression ⢠environmental cigarette smoke SpragueâDawley rats (Harlan Italy, Correzzana, Milan, Italy), weighing 315-320 g, were either exposed to ECS for 4 weeks or kept in filtered air for the same period of time (sham).
Project description:While microRNAs have extensively been investigated in cancer research, little is known regarding their response to noxious agents in apparently healthy tissues. We analyzed the expression of 484 miRNAs in the lung of rats exposed to environmental cigarette smoke (ECS) for 28 days. ECS downregulated 126 miRNAs (26.0%) at least two-fold and 24 miRNAs more than three-fold. We previously demonstrated that 107 of 4858 genes (2.9%) and 50 of 518 proteins (9.7%) were upregulated by ECS in the same tissue, consistently with the role of microRNAs as negative regulators of gene expression. The most remarkably downregulated microRNAs belonged to the families of let-7, miR-10, miR-26, miR-30, miR-34, miR-99, miR- 122, miR-123, miR-124, miR-125, miR-140, miR-145, miR-146, miR-191, miR-192, miR-219, miR-222, and miR-223, which regulate stress response, apoptosis, proliferation, angiogenesis, and expression of genes. In contrast, miR-294, an inhibitor of transcriptional repressor genes, was upregulated by ECS. There was a strong parallelism in dysregulation of rodent microRNAs and their human homologues, which are often transcribed from genes localized in fragile sites deleted in lung cancer. Five ECS-downregulated microRNAs are known to be affected by single nucleotide polymorphisms. Thus, changes in microRNA expression are an early event following exposure to cigarette smoke. Keywords: microRNAs • gene expression • environmental cigarette smoke