Cellular localization of NRF2 determines the self-renewal and osteogenic differentiation potential of human MSCs via the P53–SIRT1 axis

Abstract

NRF2 (nuclear factor erythroid-derived 2-like 2) plays an important role in defense against oxidative stress at the cellular level. Recently, the roles of NRF2 in embryonic and adult stem cells have been reported, but its role in maintaining self-renewal and differentiation potential remains unknown. We studied the mechanisms of NRF2 action in mesenchymal stem cells (MSCs) derived from human bone marrow. We found that the cellular localization of NRF2 changed during prolonged cell passage and osteogenic differentiation. Blocking the nuclear import of NRF2 using ochratoxin A (OTA) induced the loss of the self-renewal and osteogenic potential of early-passage (EP) MSCs. Conversely, reinforcing the nuclear import of NRF2 using tert-butylhydroquinone (t-BHQ) improved the self-renewal capacity and maintained the differentiation potential in the osteogenic lineage of EP MSCs. Real-time quantitative PCR and western blot analysis showed that NRF2 positively regulates sirtuin 1 (SIRT1) at the mRNA and protein levels via the negative regulation of p53. The self-renewal and osteogenic potential suppressed in OTA-treated or NRF2-targeting small hairpin RNA (shRNA)-infected EP MSCs were rescued by introducing small interfering RNA (siRNA) targeting p53. t-BHQ treatment in late-passage (LP) MSCs, which lost their self-renewal and osteogenic potential, reversed these effects. In LP MSCs treated with t-BHQ for ∼7 days, the phosphorylation and nuclear localization of NRF2 improved and SIRT1 protein level increased, whereas p53 protein levels decreased. Therefore, our results suggest that NRF2 plays an important role in regulating p53 and SIRT1 to maintain MSC stemness. This study is the first to establish a functional link between NRF2 and SIRT1 expression in the maintenance of MSC self-renewal and differentiation potential.

C1q/Tumor Necrosis Factor–Related Protein-9 Regulates the Fate of Implanted Mesenchymal Stem Cells and Mobilizes Their Protective Effects Against Ischemic Heart Injury via Multiple Novel Signaling Pathways

“…Nrf2 (nuclear factor erythroid-derived 2-like 2) importantly defends against oxidative stress at the cellular level 19 . Expression of Sod-2, Sod-3, HO-1, and Prdx1 are regulated by Nrf2 20, 21 .…”

SIRT1 Alleviates LPS-Induced IL-1β Production by Suppressing NLRP3 Inflammasome Activation and ROS Production in Trophoblasts

“…Previous studies have shown bidirectional crosstalk between SIRT1 and nuclear factor erythroid 2-related factor 2 (Nrf2) in human renal proximal tubular and glomerular mesangial cells [57,58]. The upregulation of SIRT1 activates Nrf2 through the inhibition of p53 or activation of AMPK [59][60][61]. In addition, it is known that in ischemic injury Nrf2 suppresses NLRP3 inflammasome activation through regulating the thioredoxin-interacting protein (TXNIP) complex [62].…”

Role of NF-E2 related factor 2 (Nrf2) on chemotherapy resistance in acute myeloid leukemia (AML) and the effect of pharmacological inhibition of Nrf2

“…No: TRCN0000007555) was cloned into Plko.1 Tet-ON plasmid (Addgene clone ID 21915). This shRNA showed significantly better knock down of Nrf2 as reported previously in recent studies [28,29]. For generating Nrf2 overexpression plasmids, a lentiviral vector containing full-length Nrf2 cDNA, PLX304-Nrf2 (clone ID HsCD00445044), was purchased from DNASU plasmid repository (Biodesign Institute, Arizona State University, Arizona, USA).…”