Suppression of transient receptor potential melastatin 7 regulates pluripotency, proliferation, and differentiation of mouse embryonic stem cells via mechanistic target of rapamycin‐extracellular signal‐regulated kinase activation

• Published in: Journal of cellular biochemistry Vol. 123; no. 3; pp. 547 - 567
• Main Authors: Kim, Wansoo, Park, Song, Kwon, Wookbong, Kim, Daehwan, Park, Jin‐Kyu, Han, Jee Eun, Cho, Gil‐Jae, Han, Se‐Hyeon, Sung, Yonghun, Yi, Jun‐Koo, Kim, Myoung Ok, Ryoo, Zae Young, Choi, Seong‐Kyoon
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01-03-2022
• Subjects: Animals; Apoptosis; Calcium (intracellular); Calcium ions; Cell activation; Cell cycle; Cell Differentiation; Cell Proliferation; Cell self-renewal; Differentiation; Embryo cells; Embryogenesis; Embryonic growth stage; ERK; Extracellular signal-regulated kinase; Extracellular Signal-Regulated MAP Kinases - metabolism; G1 phase; Ion channels; Kinases; mESC; mESC differentiation; Mice; Mouse Embryonic Stem Cells - metabolism; mTOR; Oct-4 protein; Phosphorylation; Pluripotency; Rapamycin; Receptors; RNA, Small Interfering - metabolism; Sirolimus; Stem cell transplantation; Stem cells; Teratoma; TOR protein; TOR Serine-Threonine Kinases - genetics; TOR Serine-Threonine Kinases - metabolism; Transcription; Transient receptor potential proteins; TRPM Cation Channels - genetics; TRPM Cation Channels - metabolism; Trpm7; mTOR; Trpm7; mESC; pluripotency; mESC differentiation; ERK
• ISSN: 0730-2312; 1097-4644
• DOI: 10.1002/jcb.30199

Mouse embryonic stem cells (mESCs) are a widely used model for their diverse availability in studying early embryonic development and their application in regenerative treatment of various intractable diseases. Transient receptor potential melastatin 7 (Trpm7) regulates Ca2+ as a nonselective ion channel and is essential for early embryonic development; however, the precise role of Trpm7 in mESCs has not been clearly elucidated. In this study, we showed that the inhibition of Trpm7 affects the pluripotency and self‐renewal of mESCs. We found that short hairpin RNA (shRNA)‐mediated suppression of Trpm7 resulted in decreased expression of transcriptional regulators, Oct4 and Sox2, which maintain stemness in mESCs. In addition, Trpm7 knockdown led to alterations in the basic properties of mESCs, such as decreased proliferation, cell cycle arrest at the G0/G1 phase, and increased apoptosis. Furthermore, embryoid body (EB) formation and teratoma formation assays revealed abnormal regulation of differentiation due to Trpm7 knockdown, including the smaller size of EBs, elevated ectodermal differentiation, and diminished endodermal and mesodermal differentiation. We found that EB Day 7 samples displayed decreased intracellular Ca2+ levels compared to those of the scrambled group. Finally, we identified that these alterations induced by Trpm7 knockdown occurred due to decreased phosphorylation of mechanistic target of rapamycin (mTOR) and subsequent activation of extracellular signal‐regulated kinase (ERK) in mESCs. Our findings suggest that Trpm7 could be a novel regulator for maintaining stemness and modulating the differentiation of mESCs.