Enhanced PRL-1 expression in placenta-derived mesenchymal stem cells accelerates hepatic function via mitochondrial dynamics in a cirrhotic rat model

Abstract

Background: Placenta-derived mesenchymal stem cells (PD-MSCs) have been highlighted as an alternative cell therapy agent that has become a next-generation stem cell treatment. Phosphatase of regenerating liver-1 (PRL-1), an immediate early gene, plays a critical role during liver regeneration. Here, we generated enhanced PRL-1 in PD-MSCs (PD-MSCsPRL-1, PRL-1+) using lentiviral and nonviral gene delivery systems and investigated mitochondrial functions by PD-MSCPRL-1 transplantation for hepatic functions in a rat bile duct ligation (BDL) model. Methods: PD-MSCsPRL-1 were generated by lentiviral and nonviral AMAXA gene delivery systems and analyzed for their characteristics and mitochondrial metabolic functions. Liver cirrhosis was induced in Sprague-Dawley (SD) rats using common BDL for 10 days. PKH67+ naïve and PD-MSCsPRL-1 were intravenously administered into cirrhotic rats using a nonviral system (2x106 cells/animal). The animals were sacrificed at 1, 2, 3, and 5 weeks after transplantation and engraftment of stem cells, and histopathological analysis and hepatic mitochondrial functions were performed. Results: PD-MSCsPRL-1 were successfully generated using lentiviral and nonviral AMAXA systems and maintained characteristics similar to those of naïve cells. Compared with naïve cells, PD-MSCsPRL-1 improved respirational metabolic states of mitochondria. In particular, mitochondria in PD-MSCsPRL-1 generated by the nonviral AMAXA system showed a significant increase in the respirational metabolic state, including ATP production and mitochondrial biogenesis (*p<0.05). Furthermore, transplantation of PD-MSCsPRL-1 using a nonviral AMAXA system promoted engraftment into injured target liver tissues of a rat BDL cirrhotic model and enhanced the metabolism of mitochondria via increased mtDNA and ATP production, thereby improving therapeutic efficacy. Conclusions: Our findings will further our understanding of the therapeutic mechanism of enhanced MSCs and provide useful data for the development of next-generation MSC-based cell therapy and therapeutic strategies for regenerative medicine in liver disease.

Activation of the EGFR-PI3K-CaM Pathway by PRL-1 Ameliorates Liver Cirrhosis via ER Stress-dependent Calcium

“…However, hepatocyte-speci c calcium channels in the ER and mitochondria need to be further con rmed for isolation from the ER and mitochondria in hepatocytes. Our previous reports demonstrated that functionenhanced PD-MSCs PRL−1 were generated [38] and that transplantation improved mitochondrial biogenesis and liver function in a BDL-injured rat model [20]. Additionally, PD-MSCs PRL−1 promoted antiadipogenic effects via improved IGFBP expression in orbital broblasts [39].…”

“…Hepatic regenerative effects of PD-MSCs PRL-1 in a rat model of BDL Previously, we reported that administration of PD-MSCs PRL−1 improved hepatic functions in a rat model of BDL [20]. In the NTx group, the collagen accumulation in liver tissues was signi cantly increased compared to that in the normal group.…”

“…In previous reports, we demonstrated that placenta-derived MSC (PD-MSC) transplantation enhances hepatic function through increased anti brotic effects, proliferation, and autophagy in a chronic liver disease model [19]. Also, PRL-1-overexpressing PD-MSCs (PD-MSCs PRL-1 ) enhanced hepatic functions by increasing mitochondrial functions and proliferative potential in a chronic liver disease model compared to naïve PD-MSCs [20]. However, whether PD-MSC PRL-1 transplantation is related to the ER stress mechanism induced by calcium in cirrhotic livers remains unclear.…”

“…The PRL-1 plasmid vector was purchased from Origene (#RG200435; Rockville, MD, USA). For overexpression of the PRL-1 gene, naïve PD-MSCs (passage = 7) were transfected using the AMAXA nucleofector system (Lonza, Basel, Switzerland) according to the manufacturer's instructions as previously described [20].…”