Mesenchymal Stem Cells Reduce Corneal Fibrosis and Inflammation via Extracellular Vesicle-Mediated Delivery of miRNA

Shojaati, Golnar; Khandaker, Irona; Funderburgh, Martha L.; Mann, Mary M.; Basu, Rohan; Stolz, Donna B.; Geary, Moira L.; Santos, Aurélie Dos; Deng, Sophie X.; Funderburgh, James L.

doi:10.1002/sctm.18-0297

Cited by 125 publications

(118 citation statements)

References 63 publications

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“…The production of TSG-6 (tumor necrosis factor-stimulated gene 6) in wounded corneas further demonstrated the ability of hCSSC to block neutrophil infiltration, thus reducing the myofibroblast differentiation and scar-associated gene expression, including collagen 3a1 (Col3a1), tenascin C (TNC) and α-smooth muscle actin (αSMA) [ 18 ]. Recently, we have shown that hCSSC induced corneal regeneration via an indirect cell-cell interaction, an effect consistent with the paracrine action of extracellular vesicles [ 19 ]. Such treatment effects disappeared when the vesicles were depleted of microRNAs, after specific Alix knockdown, suggesting that some microRNAs could have therapeutic effects on corneal fibrosis and stromal regeneration.…”

Section: Introductionmentioning

confidence: 66%

“…In our previous analysis using nanoString™ for Mouse Inflammation and Mouse Pan-Cancer Panels, a number of mouse inflammatory genes were affected by hCSSC treatment [ 19 ]. We assayed the RNA expression of early inflammatory genes, including CD80, C-X-C motif chemokine ligand 5 (CXCL5), lipocalin 2 (Lcn2), plasminogen activator urokinase receptor (PLAUR), pro-platelet basic protein (Ppbp) and secreted phosphoprotein 1 ( Spp1 ).…”

Section: Resultsmentioning

confidence: 99%

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The anti-scarring effect of corneal stromal stem cell therapy is mediated by transforming growth factor β3

et al. 2020

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Background Corneal stromal stem cells (CSSC) reduce corneal inflammation, prevent fibrotic scarring, and regenerate transparent stromal tissue in injured corneas. These effects rely on factors produced by CSSC to block the fibrotic gene expression. This study investigated the mechanism of the scar-free regeneration effect. Methods Primary human CSSC (hCSSC) from donor corneal rims were cultivated to passage 3 and co-cultured with mouse macrophage RAW264.7 cells induced to M1 pro-inflammatory phenotype by treatment with interferon-γ and lipopolysaccharides, or to M2 anti-inflammatory phenotype by interleukin-4, in a Transwell system. The time-course expression of human transforming growth factor β3 (hTGFβ3) and hTGFβ1 were examined by immunofluorescence and qPCR. TGFβ3 knockdown for > 70% in hCSSC [hCSSC-TGFβ3(si)] was achieved by small interfering RNA transfection. Naïve CSSC and hCSSC-TGFβ3(si) were transplanted in a fibrin gel to mouse corneas, respectively, after wounding by stromal ablation. Corneal clarity and the expression of mouse inflammatory and fibrosis genes were examined. Results hTGFβ3 was upregulated by hCSSC when co-cultured with RAW cells under M1 condition. Transplantation of hCSSC to wounded mouse corneas showed significant upregulation of hTGFβ3 at days 1 and 3 post-injury, along with the reduced expression of mouse inflammatory genes (CD80, C-X-C motif chemokine ligand 5, lipocalin 2, plasminogen activator urokinase receptor, pro-platelet basic protein, and secreted phosphoprotein 1). By day 14, hCSSC treatment significantly reduced the expression of fibrotic and scar tissue genes (fibronectin, hyaluronan synthase 2, Secreted protein acidic and cysteine rich, tenascin C, collagen 3a1 and α-smooth muscle actin), and the injured corneas remained clear. However, hCSSC-TGFβ3(si) lost these anti-inflammatory and anti-scarring functions, and the wounded corneas showed intense scarring. Conclusion This study has demonstrated that the corneal regenerative effect of hCSSC is mediated by TGFβ3, inducing a scar-free tissue response.

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Section: Introductionmentioning

confidence: 66%

Section: Resultsmentioning

confidence: 99%

The anti-scarring effect of corneal stromal stem cell therapy is mediated by transforming growth factor β3

et al. 2020

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“…83,84 Several other gene knockdown strategies for ocular scarring are also being tested in animals. 1,[9][10][11]14 In this study, we used the fully modified siRNA conjugate to achieve maximal activity and longevity of the effect in vivo. 48 Regarding the possibility of off-target effects, the asymmetric (20/15) chemically modified siRNA used in this study was designed to avoid any sequence identity with other rabbit genes within the 2-18 region of the anti-sense strand.…”

Section: Discussionmentioning

confidence: 99%

“…3,4 Mitomycin C (MMC) to improve healing and avert scarring is a standard of care for some of these indications, but there is a high failure rate with the filtration surgery and cell toxicity concerns in corneal surgeries. 2,[5][6][7][8] Although there are several other therapeutic modalities being tested for the cornea, including viral delivery of genes, growth factors, and stem cells derived from various sources, 1,[9][10][11][12][13][14][15] currently transplant of non-autologous corneal tissue is the only option available. Furthermore, there is a global shortage of tissue and limited access to this procedure for most of the world.…”

Section: Introductionmentioning

confidence: 99%

USP10 Targeted Self-Deliverable siRNA to Prevent Scarring in the Cornea

Boumil

Castro

Phillips

et al. 2020

Molecular Therapy - Nucleic Acids

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Ocular scarring after surgery, trauma, or infection leads to vision loss. The transparent cornea is an excellent model system to test anti-scarring therapies. Cholesterol-conjugated fully modified asymmetric small interfering RNAs (siRNAs) (selfdeliverable siRNAs [sdRNAs]) are a novel modality for in vivo gene knockdown, transfecting cells and tissues without any additional formulations. Myofibroblasts are a main contributor to scarring and fibrosis. a v integrins play a central role in myofibroblast pathological adhesion, overcontraction, and transforming growth factor b (TGF-b) activation. Previously, we demonstrated that a v integrins are protected from intracellular degradation after wounding by upregulation of the deubiquitinase (DUB) ubiquitin-specific protease 10 (USP10), leading to integrin cell surface accumulation. In this study, we tested whether knockdown of USP10 with a USP10-targeting sdRNA (termed US09) will reduce scarring after wounding a rabbit cornea in vivo. The wounded corneal stroma was treated once with US09 or non-targeting control (NTC) sdRNA. At 6 weeks US09 treatment resulted in faster wound closure, limited scarring, and suppression of fibrotic markers and immune response. Specifically, fibronectin-extra domain A (EDA), collagen III, and a-smooth muscle actin (p < 0.05), CD45 + cell infiltration (p < 0.01), and apoptosis at 24 (p < 0.01) and 48 h (p < 0.05) were reduced post-wounding. Corneal thickness and cell proliferation were restored to unwounded parameters. Targeting the DUB, USP10 is a novel strategy to reduce scarring. This study indicates that ubiquitin-mediated pathways should be considered in the pathogenesis of fibrotic healing.

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“…Corneal epithelial layer was recovered over the damaged tissue after 24 h, and corneal scarring was prevented in treated eyes after 2 weeks. It was concluded that EVs eliminated the requirement of live cells which could be safer and cheaper for clinical trials [171].…”

Section: 222mentioning

confidence: 99%

Bioengineering Approaches for Corneal Regenerative Medicine

Mahdavi

Abdekhodaie

Mashayekhan

et al. 2020

Tissue Eng Regen Med

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BACKGROUND: Since the cornea is responsible for transmitting and focusing light into the eye, injury or pathology affecting any layer of the cornea can cause a detrimental effect on visual acuity. Aging is also a reason for corneal degeneration. Depending on the level of the injury, conservative therapies and donor tissue transplantation are the most common treatments for corneal diseases. Not only is there a lack of donor tissue and risk of infection/rejection, but the inherent ability of corneal cells and layers to regenerate has led to research in regenerative approaches and treatments. METHODS: In this review, we first discussed the anatomy of the cornea and the required properties for reconstructing layers of the cornea. Regenerative approaches are divided into two main categories; using direct cell/growth factor delivery or using scaffold-based cell delivery. It is expected delivered cells migrate and integrate into the host tissue and restore its structure and function to restore vision. Growth factor delivery also has shown promising results for corneal surface regeneration. Scaffold-based approaches are categorized based on the type of scaffold, since it has a significant impact on the efficiency of regeneration, into the hydrogel and non-hydrogel based scaffolds. Various types of cells, biomaterials, and techniques are well covered. RESULTS: The most important characteristics to be considered for biomaterials in corneal regeneration are suitable mechanical properties, biocompatibility, biodegradability, and transparency. Moreover, a curved shape structure and spatial arrangement of the fibrils have been shown to mimic the corneal extracellular matrix for cells and enhance cell differentiation. CONCLUSION: Tissue engineering and regenerative medicine approaches showed to have promising outcomes for corneal regeneration. However, besides proper mechanical and optical properties, other factors such as appropriate sterilization method, storage, shelf life and etc. should be taken into account in order to develop an engineered cornea for clinical trials.

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Mesenchymal Stem Cells Reduce Corneal Fibrosis and Inflammation via Extracellular Vesicle-Mediated Delivery of miRNA

Cited by 125 publications

References 63 publications

The anti-scarring effect of corneal stromal stem cell therapy is mediated by transforming growth factor β3

The anti-scarring effect of corneal stromal stem cell therapy is mediated by transforming growth factor β3

USP10 Targeted Self-Deliverable siRNA to Prevent Scarring in the Cornea

Bioengineering Approaches for Corneal Regenerative Medicine

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