TLR3-overexpressing umbilical cord mesenchymal stromal cells suppress immune responses to attenuate high-risk corneal transplantation rejection

Abstract Background The success rates of high-risk corneal transplantation are significantly hindered by immunological rejection. Umbilical cord mesenchymal stem cells (UC-MSCs) have emerged as a potential solution to improve graft outcomes. Toll-like receptors (TLRs) play pivotal roles in the immun...

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Main Authors: Yaqi Cheng, Huaxin Chen, Simin Gu, Weihua Li, Huan Yu, Jianqiang Zhang, Huini Zhang, Jiayi Lin, Haocheng Zhu, Youyu Liu, Wenqiong Li, Ting Fu, Haoyu Zeng, Tao Wang, Shiqi Ling
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Language:English
Published: BMC 2025-07-01
Series:Stem Cell Research & Therapy
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Online Access:https://doi.org/10.1186/s13287-025-04510-3
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author Yaqi Cheng
Huaxin Chen
Simin Gu
Weihua Li
Huan Yu
Jianqiang Zhang
Huini Zhang
Jiayi Lin
Haocheng Zhu
Youyu Liu
Wenqiong Li
Ting Fu
Haoyu Zeng
Tao Wang
Shiqi Ling
author_facet Yaqi Cheng
Huaxin Chen
Simin Gu
Weihua Li
Huan Yu
Jianqiang Zhang
Huini Zhang
Jiayi Lin
Haocheng Zhu
Youyu Liu
Wenqiong Li
Ting Fu
Haoyu Zeng
Tao Wang
Shiqi Ling
author_sort Yaqi Cheng
collection DOAJ
description Abstract Background The success rates of high-risk corneal transplantation are significantly hindered by immunological rejection. Umbilical cord mesenchymal stem cells (UC-MSCs) have emerged as a potential solution to improve graft outcomes. Toll-like receptors (TLRs) play pivotal roles in the immune response, however, their specific functions in UC-MSC-based treatments for corneal graft rejection requires further investigation. Methods New Zealand rabbits that underwent high-risk corneal transplantation were treated with UC-MSC-coated contact lenses (MSCohi-O), while blank lenses and untreated grafts served as controls. Clinical manifestations related to rejection were assessed. RNA-seq was performed on the grafts. GSEA, CIBERSORT, and flow cytometry analyses were performed to investigate the activation of signaling pathways and immune changes. The microarray dataset GSE68610, which contains transcriptome data for normal human UC-MSCs and UC-MSCs treated with cytokines, was analyzed to evaluate TLR expression and identify the key factors involved in the anti-inflammatory effect of UC-MSCs. Overexpression and knockout of TLR3 were performed in UC-MSCs, and the therapeutic effects of TLR3-activated and -inactivated UC-MSCs were compared in a high-risk corneal transplantation model. Results MSCohi-O treatment alleviated corneal opacity and edema, inhibited neovascularization, promoted epithelialization, and prolonged the survival time of corneal grafts (all p < 0.05). GSEA revealed that the allograft rejection pathway was upregulated in untreated grafts and downregulated in UC-MSC-treated grafts. Increased numbers of Tregs and decreased numbers of Th17 cells were observed in UC-MSC-treated corneas. An analysis of the GSE68610 dataset revealed that TLR3 expression was upregulated in cytokine-activated UC-MSCs, suggesting that TLR3 is a potential regulator of the immunosuppression function of UC-MSCs. TLR3-overexpressing UC-MSCs exhibited enhanced suppression of rejection in high-risk corneal transplants, whereas TLR3 knockdown diminished these effects. Conclusions This study shows that the localized application of UC-MSC-coated contact lenses is capable of inhibiting rejection in high-risk corneal transplantation. TLR3-overexpressing UC-MSCs have enhanced antirejection and immunomodulatory effects. This research could offer a safer and more effective therapeutic strategy to prevent corneal transplant rejection.
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spelling doaj-art-89768cc3159e46be8ad36baf1c4cfddb2025-08-20T03:42:20ZengBMCStem Cell Research & Therapy1757-65122025-07-0116111810.1186/s13287-025-04510-3TLR3-overexpressing umbilical cord mesenchymal stromal cells suppress immune responses to attenuate high-risk corneal transplantation rejectionYaqi Cheng0Huaxin Chen1Simin Gu2Weihua Li3Huan Yu4Jianqiang Zhang5Huini Zhang6Jiayi Lin7Haocheng Zhu8Youyu Liu9Wenqiong Li10Ting Fu11Haoyu Zeng12Tao Wang13Shiqi Ling14Department of Ophthalmology, The Third Affiliated Hospital, Sun Yat-sen UniversityBiotherapy Centre, The Third Affiliated Hospital, Sun Yat-sen UniversityDepartment of Ophthalmology, The Third Affiliated Hospital, Sun Yat-sen UniversityState Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Sun Yat- sen UniversityState Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Sun Yat- sen UniversityDepartment of Ophthalmology, The Third Affiliated Hospital, Sun Yat-sen UniversityDepartment of Ophthalmology, The Third Affiliated Hospital, Sun Yat-sen UniversityDepartment of Ophthalmology, The Third Affiliated Hospital, Sun Yat-sen UniversityDepartment of Ophthalmology, The Third Affiliated Hospital, Sun Yat-sen UniversityGuangdong Procapzoom Biosciences Co., LtdGuangdong Procapzoom Biosciences Co., LtdGuangdong Procapzoom Biosciences Co., LtdGuangdong Procapzoom Biosciences Co., LtdDepartment of Ophthalmology, The Third Affiliated Hospital, Sun Yat-sen UniversityDepartment of Ophthalmology, The Third Affiliated Hospital, Sun Yat-sen UniversityAbstract Background The success rates of high-risk corneal transplantation are significantly hindered by immunological rejection. Umbilical cord mesenchymal stem cells (UC-MSCs) have emerged as a potential solution to improve graft outcomes. Toll-like receptors (TLRs) play pivotal roles in the immune response, however, their specific functions in UC-MSC-based treatments for corneal graft rejection requires further investigation. Methods New Zealand rabbits that underwent high-risk corneal transplantation were treated with UC-MSC-coated contact lenses (MSCohi-O), while blank lenses and untreated grafts served as controls. Clinical manifestations related to rejection were assessed. RNA-seq was performed on the grafts. GSEA, CIBERSORT, and flow cytometry analyses were performed to investigate the activation of signaling pathways and immune changes. The microarray dataset GSE68610, which contains transcriptome data for normal human UC-MSCs and UC-MSCs treated with cytokines, was analyzed to evaluate TLR expression and identify the key factors involved in the anti-inflammatory effect of UC-MSCs. Overexpression and knockout of TLR3 were performed in UC-MSCs, and the therapeutic effects of TLR3-activated and -inactivated UC-MSCs were compared in a high-risk corneal transplantation model. Results MSCohi-O treatment alleviated corneal opacity and edema, inhibited neovascularization, promoted epithelialization, and prolonged the survival time of corneal grafts (all p < 0.05). GSEA revealed that the allograft rejection pathway was upregulated in untreated grafts and downregulated in UC-MSC-treated grafts. Increased numbers of Tregs and decreased numbers of Th17 cells were observed in UC-MSC-treated corneas. An analysis of the GSE68610 dataset revealed that TLR3 expression was upregulated in cytokine-activated UC-MSCs, suggesting that TLR3 is a potential regulator of the immunosuppression function of UC-MSCs. TLR3-overexpressing UC-MSCs exhibited enhanced suppression of rejection in high-risk corneal transplants, whereas TLR3 knockdown diminished these effects. Conclusions This study shows that the localized application of UC-MSC-coated contact lenses is capable of inhibiting rejection in high-risk corneal transplantation. TLR3-overexpressing UC-MSCs have enhanced antirejection and immunomodulatory effects. This research could offer a safer and more effective therapeutic strategy to prevent corneal transplant rejection.https://doi.org/10.1186/s13287-025-04510-3Mesenchymal stromal cellsCorneal transplantation rejectionTLR3Immune
spellingShingle Yaqi Cheng
Huaxin Chen
Simin Gu
Weihua Li
Huan Yu
Jianqiang Zhang
Huini Zhang
Jiayi Lin
Haocheng Zhu
Youyu Liu
Wenqiong Li
Ting Fu
Haoyu Zeng
Tao Wang
Shiqi Ling
TLR3-overexpressing umbilical cord mesenchymal stromal cells suppress immune responses to attenuate high-risk corneal transplantation rejection
Stem Cell Research & Therapy
Mesenchymal stromal cells
Corneal transplantation rejection
TLR3
Immune
title TLR3-overexpressing umbilical cord mesenchymal stromal cells suppress immune responses to attenuate high-risk corneal transplantation rejection
title_full TLR3-overexpressing umbilical cord mesenchymal stromal cells suppress immune responses to attenuate high-risk corneal transplantation rejection
title_fullStr TLR3-overexpressing umbilical cord mesenchymal stromal cells suppress immune responses to attenuate high-risk corneal transplantation rejection
title_full_unstemmed TLR3-overexpressing umbilical cord mesenchymal stromal cells suppress immune responses to attenuate high-risk corneal transplantation rejection
title_short TLR3-overexpressing umbilical cord mesenchymal stromal cells suppress immune responses to attenuate high-risk corneal transplantation rejection
title_sort tlr3 overexpressing umbilical cord mesenchymal stromal cells suppress immune responses to attenuate high risk corneal transplantation rejection
topic Mesenchymal stromal cells
Corneal transplantation rejection
TLR3
Immune
url https://doi.org/10.1186/s13287-025-04510-3
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