Protective effect of <i>GDNF</i>‐engineered amniotic fluid‐derived stem cells on the renal ischaemia reperfusion injury in vitro

Wang, Jia; Wang, Fengzhen; Wang, Zhuojun; Li, Shulin; Chen, Lu; Liu, Caixia; Sun, Dong

doi:10.1111/cpr.12400

Cited by 14 publications

(12 citation statements)

References 41 publications

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“…Accumulating evidence indicates that MSCs attenuate renal injury and dysfunction in several animal models, and their efficacy in patients with renal disease is currently being tested in clinical trials 38 , 39 . We previously showed a reduced level of cell apoptosis in a hypoxia-reoxygenation model of GDNF-engineered amniotic fluid-derived stem cells cocultured with mouse renal tubular epithelial cells in a Transwell system 40 . The current study extends our previous observations, demonstrating that UUO kidney structure and function are preserved 7 days after the delivery of GDNF-AMSC-exos.…”

Section: Discussionmentioning

confidence: 97%

Exosomes derived from GDNF-modified human adipose mesenchymal stem cells ameliorate peritubular capillary loss in tubulointerstitial fibrosis by activating the SIRT1/eNOS signaling pathway

Chen

Wang

et al. 2020

Theranostics

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Mesenchymal stem cells (MSCs) have emerged as ideal cell-based therapeutic candidates for the structural and functional restoration of the diseased kidney. Glial cell line-derived neurotrophic factor (GDNF) has been demonstrated to promote the therapeutic effect of MSCs on ameliorating renal injury. The mechanism may involve the transfer of endogenous molecules via paracrine factors to salvage injured cells, but these factors remain unknown. Methods: GDNF was transfected into human adipose mesenchymal stem cells via a lentiviral transfection system, and exosomes were isolated (GDNF-AMSC-exos). Using the unilateral ureteral obstruction (UUO) mouse model and human umbilical vein endothelial cells (HUVECs) against hypoxia/serum deprivation (H/SD) injury models, we investigated whether GDNF-AMSC-exos ameliorate peritubular capillary (PTC) loss in tubulointerstitial fibrosis and whether this effect is mediated by the Sirtuin 1 (SIRT1) signaling pathway. Additionally, by using SIRT1 activators or siRNAs, the roles of the candidate mRNA and its downstream gene in GDNF-AMSC-exo-induced regulation of endothelial cell function were assessed. PTC characteristics were detected by fluorescent microangiography (FMA) and analyzed by the MATLAB software. Results: The green fluorescent PKH67-labeled exosomes were visualized in the UUO kidneys and colocalized with CD81. GDNF-AMSC-exos significantly decreased PTC rarefaction and renal fibrosis scores in mice with UUO. In vitro studies revealed that GDNF-AMSC-exos exerted cytoprotective effects on HUVECs against H/SD injury by stimulating migration and angiogenesis as well as conferring apoptosis resistance. Mechanistically, GDNF-AMSC-exos enhanced SIRT1 signaling, which was accompanied by increased levels of phosphorylated endothelial nitric oxide synthase (p-eNOS). We also confirmed the SIRT1-eNOS interaction in HUVECs by immunoprecipitation. Furthermore, we observed a correlation of the PTC number with the SIRT1 expression level in the kidney in vivo . Conclusion: Our study unveiled a mechanism by which exosomes ameliorate renal fibrosis: GDNF-AMSC-exos may activate an angiogenesis program in surviving PTCs after injury by activating the SIRT1/eNOS signaling pathway.

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Section: Discussionmentioning

confidence: 97%

Exosomes derived from GDNF-modified human adipose mesenchymal stem cells ameliorate peritubular capillary loss in tubulointerstitial fibrosis by activating the SIRT1/eNOS signaling pathway

Chen

Wang

et al. 2020

Theranostics

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“…Glial cell line-derived neurotrophic factor (GDNF) is known for its neurorestorative and neuroprotective effects in various pathologies, including Parkinson's disease [1–3], Alzheimer's disease [4, 5], and ischaemic damage [5–8], to the central and peripheral nervous systems. Despite numerous studies confirming the neuroprotective effect of GDNF, some preclinical and clinical data suggest that increasing GDNF concentrations do not always lead to significant long-term improvements [9, 10].…”

Section: Introductionmentioning

confidence: 99%

Intracellular Neuroprotective Mechanisms in Neuron-Glial Networks Mediated by Glial Cell Line-Derived Neurotrophic Factor

Митрошина

Mishchenko

Shirokova

et al. 2019

Oxidative Medicine and Cellular Longevity

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Glial cell line-derived neurotrophic factor (GDNF) has a pronounced neuroprotective effect in various nervous system pathologies, including ischaemic brain damage and neurodegenerative diseases. In this work, we studied the effect of GDNF on the ultrastructure and functional activity of neuron-glial networks during acute hypoxic exposure, a key damaging factor in numerous brain pathologies. We analysed the molecular mechanisms most likely involved in the positive effects of GDNF. Hypoxia modelling was performed on day 14 of culturing primary hippocampal cells obtained from mouse embryos (E18). GDNF (1 ng/ml) was added to the culture medium 20 min before oxygen deprivation. Acute hypoxia-induced irreversible changes in the ultrastructure of neurons and astrocytes led to the loss of functional Сa2+ activity and neural network disruption. Destructive changes in the mitochondrial apparatus and its functional activity characterized by an increase in the basal oxygen consumption rate and respiratory chain complex II activity during decreased stimulated respiration intensity were observed 24 hours after hypoxic injury. At a concentration of 1 ng/ml, GDNF maintained the functional metabolic network activity in primary hippocampal cultures and preserved the structure of the synaptic apparatus and number of mature chemical synapses, confirming its neuroprotective effect. GDNF maintained the normal structure of mitochondria in neuronal outgrowth but not in the soma. Analysis of the possible GDNF mechanism revealed that RET kinase, a component of the receptor complex, and the PI3K/Akt pathway are crucial for the neuroprotective effect of GDNF. The current study also revealed the role of GDNF in the regulation of HIF-1α transcription factor expression under hypoxic conditions.

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“…However, research conducted by Pichel et al demonstrated that GDNF-/-mutant mice showed kidney agenesis and dysgenesis, suggesting that GDNF plays a critical role in the development of renal system [16]. According to our previous studies [17][18][19], we found that GDNF is a tissue morphogen effecting on stem cells, which could enhance migration and differentiation of stem cells and nally reverse the kidney injury. Therefore, we speculate that if pretreatment with the GDNF gene on AMSCs could better alleviate renal brosis by strengthening the ability of AMSCs differentiating into endothelial cells, promoting AMSCs homing to damaged kidney, enhancing angiogenesis through PI3K/AKT/ eNOS pathway, and further suppressing oxidative effect and EndMT.…”

Section: Introductionmentioning

confidence: 64%

“…Therefore, it is essential to maintain a normal tissue blood supply for the kidneys. As a common cause of CKD, capillary rarefaction results in the imbalance of local tissue oxygen supply and demand together with accumulation of waste products of metabolism, which further lead to a series of events, such as epithelial cells injury, the accumulation of reactive oxygen metabolites, as well as in ammation and oxidative stress [18,30,36] . Oxidative stress contributes to lipid peroxidation, increased hydrogen peroxide, DNA and protein damage.…”

Section: Discussionmentioning

confidence: 99%

Enhanced Renoprotective Effect of GDNF-Modified Adipose Derived Mesenchymal Stem Cells on Renal Interstitial Fibrosis

Wang

et al. 2020

Preprint

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Background: The therapeutic effect of Mesenchymal stem cells (MSCs) from human adipose tissue on renal interstitial fibrosis has been demonstrated by several groups. However, the way to enhance the renoprotective effect of Adipose Derived Mesenchymal Stem Cells (AMSCs) and the possible mechanisms, are still unclear. The present study aimed to determine whether glial cell line-derived neurotrophic factor (GDNF)-modified AMSCs holds an enhanced protective effect on renal fibrosis. Methods: AMSCs were isolated and purified for culture. GDNF gene was constructed to transfect into AMSCs. The ability of AMSCs and GDNF-AMSCs supernatants to promote tube formation of endothelial cells, repair damaged endothelial cell junctions and improve endothelial cell function was compared by tube formation assay, immunofluorescence techniques and vascular ring assay, respectively. Furthermore, HE and Masson staining were used to observe the histological morphology of the kidney in vivo. Peritubular Capillary Changes were detected and analysed by Fluorescence Micro-angiography (FMA). Meanwhile, the endothelial cell, hypoxia, oxidative stress, fibrotic markers and PI3K/Akt pathway proteins were measured by western blot or qRT-PCR technics. Results: Studies in vitro demonstrated that compared with AMSCs only, GDNF-AMSCs could better repair injured endothelial cells and promote angiogenesis through secreting more growth factors in the supernatant of GDNF-AMSCs culture media. Studies in vivo, unilateral ureteral obstruction (UUO)-induced fibrotic mice were injected with transfected AMSCs through their tail veins. We showed that enhanced homing of AMSCs were observed in the GDNF-AMSC group compared with the AMSC group. GDNF-AMSCs treated animals exhibited better improvement of capillary rarefaction and fibrosis induced by obstructed kidney compared with the AMSC group. Furthermore, we reported that GDNF-AMSCs protect renal tissues against microvascular injuries via activation of the PI3K/Akt signaling pathway. Therefore, GDNF-AMSCs further ameliorated the tissue hypoxia, suppressed oxidative stress, and finally inhibited endothelial to mesenchymal transition noting by decreased coexpression of endothelial cell (CD31) and myofibroblast (a-SMA) markers. Conclusion: Collectively, our data indicated that the GDNF gene enhances the ability of AMSCs in improving renal microcirculation through PI3K/Akt/eNOS signaling pathway, and afterward inhibit the EndMT process and kidney fibrogenesis, which should have wide implications in designing future remedies for chronic kidney disease (CKD) treatment.

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Protective effect of GDNF‐engineered amniotic fluid‐derived stem cells on the renal ischaemia reperfusion injury in vitro

Abstract: These data indicated that GDNF-AFSCs are beneficial to repairing damaged mRTECs significantly in vitro, which suggests GDNF-AFSCs provide new hopes of innovative interventions in different kidney disease.

Cited by 14 publications

References 41 publications

Exosomes derived from GDNF-modified human adipose mesenchymal stem cells ameliorate peritubular capillary loss in tubulointerstitial fibrosis by activating the SIRT1/eNOS signaling pathway

Exosomes derived from GDNF-modified human adipose mesenchymal stem cells ameliorate peritubular capillary loss in tubulointerstitial fibrosis by activating the SIRT1/eNOS signaling pathway

Intracellular Neuroprotective Mechanisms in Neuron-Glial Networks Mediated by Glial Cell Line-Derived Neurotrophic Factor

Enhanced Renoprotective Effect of GDNF-Modified Adipose Derived Mesenchymal Stem Cells on Renal Interstitial Fibrosis

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