Chorionic Villous Mesenchymal Stem Cells Derived Exosomes Deliver miR-135b-5p to Trophoblasts, Promoting their Proliferation and Invasion by Targeting TXNIP via β-Catenin Pathway
Background: Human chorionic villous mesenchymal stem cells (CV-MSCs) are found to be a promising and effective treatment for tissue injury. Trophoblast dysfunction during pregnancies is significantly involved in the pathogenesis of preeclampsia (PE). This work was to understand how CV-MSCs regulated trophoblast function.
Methods: In this study, we treated trophoblasts with CV-MSC-derived exosomes and RNA-seq analysis was used to understand the changes in trophoblasts. We examined the levels of TXNIP and β-catenin in trophoblasts by immunohistochemistry, western blot and qRT-PCR assays. Luciferase reporter assays and qRT-PCR assays were used to understand the role of miR135b-5p in the effects of CV-MSC-derived exosomes. The growth and invasion of trophoblasts was evaluated with the CCK-8 and transwell assays.
Results: The treatment markedly enhanced the trophoblast proliferation and invasion. Furthermore, a significant decrease of TXNIP expression and inactivation of the β-catenin pathway in CV-MSCs exosomes-treated trophoblasts was observed. Consistent with these findings, TXNIP inhibition exhibited the same effect of promoting trophoblast proliferation and invasion as induced by CV-MSC-derived exosomes, also with the accompaniment of inactivation of β-catenin pathway. In addition, overexpression of TXNIP activated the β-catenin pathway in trophoblasts, and reduced the proliferation and invasion of trophoblasts. Importantly, miR135b-5p was found to be highly expressed in CV-MSC exosomes and interact with TXNIP. The miR-135b-5p overexpression significantly elevated the proliferation and invasion of trophoblasts, which could be attenuated by TXNIP overexpression.
Conclusion: Our results suggest that TXNIP-dependent β-catenin pathway inactivation mediated by miR135b-5p which is delivered by CV-MSC-derived exosomes could promote the proliferation and invasion of trophoblasts.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
This is a list of supplementary files associated with this preprint. Click to download.
Figure S1 Characterization of primary CV-MSCs derived from human placenta tissues. (a) Representative photomicrographs of primary human CV-MSCs and PE-CV-MSCs before confluence at passage 3. The cells were examined for osteogenic and adipogenic differentiation. Scale bar = 20 μm. (b) The purity of the isolated CV-MSCs and PE-CV-MSCs was examined by flow cytometry; CV-MSCs express CD44, CD73, CD90 and CD105, with a lack of CD34, CD45, CD146, IG1 and HLA-DR expression.
FigS2. CV-MSCs derived PKH67 labeled exosomes are up taken by JAR cells after co-incubation in vitro. (a) Representative electron micrograph of exosomes isolated from VMSCs by density gradient centrifugation. Scale bar = 100nm. (b) Particle number and size analysis of CV-MSC-exo were determined by dynamic light scattering using a ZetaView® nanoparticle tracker (ParticleMetrix, Germany). (c) Western blot analysis for CD9 and Calnexin protein expression in CV-MSCs-derived exosomes and CV-MSCs. CD9 is a marker of exosomes while Calnexin is a ER marker. (d) Representative image of PKH67-labeled exosomes (green) uptake by JAR cells. Exosomes were isolated from CV-MSCs and co-cultured with JAR cells for 12 hours. DAPI (blue) were used to stain JAR cells. Scale bar = 25μm.
Posted 28 Dec, 2020
On 20 Dec, 2020
On 20 Dec, 2020
On 20 Dec, 2020
On 18 Dec, 2020
Chorionic Villous Mesenchymal Stem Cells Derived Exosomes Deliver miR-135b-5p to Trophoblasts, Promoting their Proliferation and Invasion by Targeting TXNIP via β-Catenin Pathway
Posted 28 Dec, 2020
On 20 Dec, 2020
On 20 Dec, 2020
On 20 Dec, 2020
On 18 Dec, 2020
Background: Human chorionic villous mesenchymal stem cells (CV-MSCs) are found to be a promising and effective treatment for tissue injury. Trophoblast dysfunction during pregnancies is significantly involved in the pathogenesis of preeclampsia (PE). This work was to understand how CV-MSCs regulated trophoblast function.
Methods: In this study, we treated trophoblasts with CV-MSC-derived exosomes and RNA-seq analysis was used to understand the changes in trophoblasts. We examined the levels of TXNIP and β-catenin in trophoblasts by immunohistochemistry, western blot and qRT-PCR assays. Luciferase reporter assays and qRT-PCR assays were used to understand the role of miR135b-5p in the effects of CV-MSC-derived exosomes. The growth and invasion of trophoblasts was evaluated with the CCK-8 and transwell assays.
Results: The treatment markedly enhanced the trophoblast proliferation and invasion. Furthermore, a significant decrease of TXNIP expression and inactivation of the β-catenin pathway in CV-MSCs exosomes-treated trophoblasts was observed. Consistent with these findings, TXNIP inhibition exhibited the same effect of promoting trophoblast proliferation and invasion as induced by CV-MSC-derived exosomes, also with the accompaniment of inactivation of β-catenin pathway. In addition, overexpression of TXNIP activated the β-catenin pathway in trophoblasts, and reduced the proliferation and invasion of trophoblasts. Importantly, miR135b-5p was found to be highly expressed in CV-MSC exosomes and interact with TXNIP. The miR-135b-5p overexpression significantly elevated the proliferation and invasion of trophoblasts, which could be attenuated by TXNIP overexpression.
Conclusion: Our results suggest that TXNIP-dependent β-catenin pathway inactivation mediated by miR135b-5p which is delivered by CV-MSC-derived exosomes could promote the proliferation and invasion of trophoblasts.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5