Invigorating human MSCs for transplantation therapy via Nrf2/DKK1 co-stimulation in a mice acute-on-chronic liver failure model
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Evaluation Summary:
Chen et al. demonstrate a pro-survival role of the NRF2/DKK1 axis in mesenchymal stem cells. Furthermore, the authors provide evidence that targeting this pathway can enhance survival in response to liver failure in vivo. These data highlight a novel signaling pathway to enhance efficacy of MSCs in promoting regeneration.
(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. The reviewers remained anonymous to the authors.)
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Abstract
Boosting stem cell resilience against an extrinsically harsh recipient environment is critical to therapeutic efficiency of stem cell-based transplantation innovations in liver disease contexts. We aimed to establish the efficacy of a transient plasmid-based preconditioning strategy to boost mesenchymal stromal cells (MSCs) capacity for anti-inflammation/antioxidant defense and paracrine actions on recipient hepatocytes. In MSCs, the master antioxidant regulator Nrf2 was found to bind directly to the antioxidant response element in the DKK1 promoter region. Activation of Nrf2 and DKK1 enhanced the anti-stress capacities of MSCs in vitro . In an acute-on-chronic liver failure (ACLF) murine model, transient co-overexpression of Nrf2 and DKK1 via plasmid transfection markedly improved MSC resilience against inflammatory and oxidative assaults, boosted MSC transplantation efficacy and promoted recipient liver regeneration because of a shift from the activation of the anti-regenerative IFN-γ/STAT1 pathway to the pro-regenerative IL-6/STAT3 pathway in the liver. Moreover, specific ablation of DKK1 receptor CKAP4 but not LRP6 in recipient hepatocytes nullified therapeutic benefits from MSC transplantation. In long-term observations, tumorigenicity was undetected in mice following transplantation of such transiently preconditioned MCSs. In conclusion, co-stimulation of Nrf2/DKK1 signaling decisively and safely improves the efficacy of human MSC-based therapies in mouse ACLF models through apparently CKAP4-dependent paracrine mechanisms.
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Evaluation Summary:
Chen et al. demonstrate a pro-survival role of the NRF2/DKK1 axis in mesenchymal stem cells. Furthermore, the authors provide evidence that targeting this pathway can enhance survival in response to liver failure in vivo. These data highlight a novel signaling pathway to enhance efficacy of MSCs in promoting regeneration.
(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. The reviewers remained anonymous to the authors.)
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Reviewer #1 (Public Review):
The authors endeavored to determine molecular pathways that could enhance the viability and function of MSCs. The authors identified the master anti-oxidant regulator NRF2 as a direct regulator of DKK1, a Wnt pathway inhibitor. Moreover, the authors demonstrate over expression of NRF2 and DKK1 ameliorates liver regeneration in a model of acute on chronic liver failure. The strengths of this study are their multi-tier approach utilizing molecular biology, genetic interventions and in vitro and vivo models. These findings have uncovered a novel signaling loop with the potential for enhancing MSC function in vivo.
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Reviewer #2 (Public Review):
This is an interesting study. It has the potential to improve the efficiency of stem cell-based therapies against human liver diseases. However, several of scientific and technical concerns need to be addressed to improve the study.
1. In Figure 1A-1C, the authors showed that Nrf2 directly suppresses the expression of DKK1 via an ARE on its promoter. However, because overexpression of Nrf2 or DKK1 both alleviates inflammation and oxidative stress in MSCs, the authors decided to overexpression both proteins simultaneously in subsequent experiments "to maximize the alleviative effects and to avoid the negative regulating loop between them". The rationale here is confusing: (1) have the authors tested whether the impact of Nrf2 overexpression on inflammation and oxidative stress is related to its …
Reviewer #2 (Public Review):
This is an interesting study. It has the potential to improve the efficiency of stem cell-based therapies against human liver diseases. However, several of scientific and technical concerns need to be addressed to improve the study.
1. In Figure 1A-1C, the authors showed that Nrf2 directly suppresses the expression of DKK1 via an ARE on its promoter. However, because overexpression of Nrf2 or DKK1 both alleviates inflammation and oxidative stress in MSCs, the authors decided to overexpression both proteins simultaneously in subsequent experiments "to maximize the alleviative effects and to avoid the negative regulating loop between them". The rationale here is confusing: (1) have the authors tested whether the impact of Nrf2 overexpression on inflammation and oxidative stress is related to its transcriptional suppression of DKK1? For example, can DKK1 overexpression rescue Nrf2 KD-induced increase of cell apoptosis and ROS in MSCs upon HNFa/H2O2 treatment? (2) Have the authors compared the impact of this co-overexpression on suppression of cell apoptosis and ROS with single overexpression to make sure that the co-overexpression is better than single overexpression? (3) Have the authors tried to combine Nrf2 OE with DKK1 KD or Nrf2 KD with DKK OE, and compared the impact of these different combinations with that of single overexpression?
2. Nrf2/DKK1 single and co-overexpression experiments (in all figures) need an empty vector control. -
