| Autor: |
Krasnova OA; Laboratory of Molecular Medicine, Institute of Cytology of the Russian Academy of Sciences, Tikhoretsky Ave. 4, 194064 St-Petersburg, Russia., Kulakova KA; Laboratory of Molecular Medicine, Institute of Cytology of the Russian Academy of Sciences, Tikhoretsky Ave. 4, 194064 St-Petersburg, Russia., Sopova JV; Laboratory of Molecular Medicine, Institute of Cytology of the Russian Academy of Sciences, Tikhoretsky Ave. 4, 194064 St-Petersburg, Russia.; Center of Transgenesis and Genome Editing, St. Petersburg State University, Universitetskaja Emb., 7/9, 199034 St-Petersburg, Russia., Smirnov EY; Laboratory of Regulation of Genes Function, Institute of Cytology of the Russian Academy of Sciences, Tikhoretsky Ave. 4, 194064 St-Petersburg, Russia., Silonov SA; Laboratory of Structural Dynamics, Stability and Folding of Proteins, Institute of Cytology of the Russian Academy of Sciences, Tikhoretsky Ave. 4, 194064 St-Petersburg, Russia., Lomert EV; Laboratory of Molecular Genetics of Tumor Cells, Institute of Cytology of the Russian Academy of Sciences, Tikhoretsky Ave. 4, 194064 St-Petersburg, Russia., Bystrova OA; Laboratory of Cell Morphology, Institute of Cytology of the Russian Academy of Sciences, Tikhoretsky Ave. 4, 194064 St-Petersburg, Russia., Martynova MG; Laboratory of Cell Morphology, Institute of Cytology of the Russian Academy of Sciences, Tikhoretsky Ave. 4, 194064 St-Petersburg, Russia., Neganova IE; Laboratory of Molecular Medicine, Institute of Cytology of the Russian Academy of Sciences, Tikhoretsky Ave. 4, 194064 St-Petersburg, Russia. |
| Abstrakt: |
G-protein-coupled receptors (GPCRs) are the largest family of cell surface receptors. They modulate key physiological functions and are required in diverse developmental processes including embryogenesis, but their role in pluripotency maintenance and acquisition during the reprogramming towards hiPSCs draws little attention. Meanwhile, it is known that more than 106 GPCRs are overexpressed in human pluripotent stem cells (hPSCs). Previously, to identify novel effectors of reprogramming, we performed a high-throughput RNA interference (RNAi) screening assay and identified adhesion GPCR, GPR123, as a potential reprogramming effector. Its role has not been explored before. Herein, by employing GPR123 RNAi we addressed the role of GPR123 for hPSCs. The suppression of GPR123 in hPSCs leads to the loss of pluripotency and differentiation, impacted colony morphology, accumulation of cells at the G2 phase of the cell cycle, and absence of the scratch closure. Application of the GPR123 RNAi at the initiation stage of reprogramming leads to a decrease in the percentage of the "true" hiPSC colonies, a drop in E-cadherin expression, a decrease in the percentage of NANOG+ nuclei, and the absence of actin cytoskeleton remodeling. Together this leads to the absence of the alkaline-phosphatase-positive hiPSCs colonies on the 18th day of the reprogramming process. Overall, these data indicate for the first time the essential role of GPR123 in the maintenance and acquisition of pluripotency. |
