Human pluripotent stem cells (hPSCs) are characterized by their abilities to self-renew and to differentiate into all cell types of the human body. PluriSIn1 is an N-acyl phenylhydrazine derivative that inhibits stearoyl-coA desaturase (SCD1), a key enzyme in oleic acid biosynthesis in hPSCs. The exposure of embryonic stem cells (ESCs) with 20μM PluriSIn1 for 16 hours resulted in a gradual and significant increase of apoptotic cells. ESCs treated with PluriSIn1 (20μM) also showed higher expression of cleaved caspase-3, a hallmark executioner of apoptosis, compared to the controls. By treating hPSCs with pancaspase inhibitor Z-VAD-FMK, PluriSIn1 (20μM)-induced cell death was significantly suppressed. Also in hPSCs, 20μM PluriSIn1 led to a ∼3.5-fold elevation in the expression of the spliced isoform sXBP1 and an increase of eIF2α phosphorylation. The incubation with PluriSIn1 (20μM) for 12 hours induced ∼30% protein synthesis attenuation in hPSCs but not in differentiated cells. Moreover, the exposure of undifferentiated hPSCs with PluriSIn1 (20μM) caused a 65% reduction in SCD1 activity as compared to DMSO-treated controls. When exposed to PluriSIn1 (20μM) for 24 hours, ~40% of the mouse embryos developed into high-quality blastocysts, while the rest stopped at the morula stage or developed into blastocysts without distinguished inner cell mass cells (ICMCs). In contrast, all DMSO-treated embryos in the control group developed into blastocysts with large and distinct ICMCs. Furthermore, subcutaneous injection of PluriSIn1-treated hPSCs in immune-comprised mice failed to generate teratoma in vivo[3]. Nanog is a marker for both cell pluripotency and tumor progression. PluriSin1 treatment at 20 µM for 1 day significantly induces the apoptosis of Nanog-positive induced pluripotent stem cells derivates (iPSD). In addition, PluriSin1 treatment at 20 µM for 4 days diminished Nanog-positive stem cells in cultured iPSD while not increasing apoptosis of iPS-derived cardiomyocytes (CM)[4].
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