Human corneal stromal stem cells express anti-fibrotic microRNA-29a and 381-5p – A robust cell selection tool for stem cell therapy of corneal scarring
Introduction: Corneal blindness due to scarring is treated with corneal transplantation. However, a global problem is the donor material shortage. Preclinical and clinical studies have shown that cell-based therapy using corneal stromal stem cells (CSSCs) suppresses corneal scarring, potentially med...
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| Format: | Article |
| Language: | English |
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Elsevier
2023-03-01
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| Series: | Journal of Advanced Research |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2090123222001205 |
| _version_ | 1797894394445037568 |
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| author | Gary Hin-Fai Yam Tianbing Yang Moira L Geary Mithun Santra Martha Funderburgh Elizabeth Rubin Yiqin Du Jose A Sahel Vishal Jhanji James L Funderburgh |
| author_facet | Gary Hin-Fai Yam Tianbing Yang Moira L Geary Mithun Santra Martha Funderburgh Elizabeth Rubin Yiqin Du Jose A Sahel Vishal Jhanji James L Funderburgh |
| author_sort | Gary Hin-Fai Yam |
| collection | DOAJ |
| description | Introduction: Corneal blindness due to scarring is treated with corneal transplantation. However, a global problem is the donor material shortage. Preclinical and clinical studies have shown that cell-based therapy using corneal stromal stem cells (CSSCs) suppresses corneal scarring, potentially mediated by specific microRNAs transported in extracellular vesicles (EVs). However, not every CSSC batch from donors achieves similar anti-scarring effects. Objectives: To examine miRNA profiles in EVs from human CSSCs showing “healing” versus “non-healing” effects on corneal scarring and to design a tool to select CSSCs with strong healing potency for clinical applications. Methods: Small RNAs from CSSC-EVs were extracted for Nanostring nCounter Human miRNA v3 assay. MicroRNAs expressed > 20 folds in “healing” EVs (P < 0.05) were subject to enriched gene ontology (GO) term analysis. MiRNA groups with predictive regulation on inflammatory and fibrotic signalling were studied by mimic transfection to (1) mouse macrophages (RAW264.7) for M1 phenotype assay; (2) human corneal keratocytes for cytokine-induced fibrosis, and (3) human CSSCs for corneal scar prevention in vivo. The expression of miR-29a was screened in additional CSSC batches and the anti-scarring effect of cells was validated in mouse corneal wounds. Results: Twenty-one miRNAs were significantly expressed in “healing” CSSC-EVs and 9 miRNA groups were predicted to associate with inflammatory and fibrotic responses, and tissue regeneration (P <10−6). Overexpression of miR-29a and 381-5p significantly prevented M1 phenotype transition in RAW264.7 cells after lipopolysaccharide treatment, suppressed transforming growth factor β1-induced fibrosis marker expression in keratocytes, and reduced scarring after corneal injury. High miR-29a expression in EV fractions distinguished human CSSCs with strong healing potency, which inhibited corneal scarring in vivo. Conclusion: We characterized the anti-inflammatory and fibrotic roles of miR-29a and 381-5p in CSSCs, contributing to scar prevention. MiR-29a expression in EVs distinguished CSSCs with anti-scarring quality, identifying good quality cells for a scarless corneal healing. |
| first_indexed | 2024-04-10T07:08:21Z |
| format | Article |
| id | doaj.art-5b9861b24c7f4e1385e9d790328a6fb3 |
| institution | Directory Open Access Journal |
| issn | 2090-1232 |
| language | English |
| last_indexed | 2024-04-10T07:08:21Z |
| publishDate | 2023-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Advanced Research |
| spelling | doaj.art-5b9861b24c7f4e1385e9d790328a6fb32023-02-27T04:07:04ZengElsevierJournal of Advanced Research2090-12322023-03-0145141155Human corneal stromal stem cells express anti-fibrotic microRNA-29a and 381-5p – A robust cell selection tool for stem cell therapy of corneal scarringGary Hin-Fai Yam0Tianbing Yang1Moira L Geary2Mithun Santra3Martha Funderburgh4Elizabeth Rubin5Yiqin Du6Jose A Sahel7Vishal Jhanji8James L Funderburgh9Department of Ophthalmology, University of Pittsburgh School of medicine, Pittsburgh, PA 15213, United States; McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA 15213, United States; Corresponding author at: Department of Ophthalmology, University of Pittsburgh, 203 Lothrop Street, Pittsburgh, PA 15213, USA.Department of Ophthalmology, University of Pittsburgh School of medicine, Pittsburgh, PA 15213, United StatesDepartment of Ophthalmology, University of Pittsburgh School of medicine, Pittsburgh, PA 15213, United StatesDepartment of Ophthalmology, University of Pittsburgh School of medicine, Pittsburgh, PA 15213, United StatesDepartment of Ophthalmology, University of Pittsburgh School of medicine, Pittsburgh, PA 15213, United StatesDepartment of Ophthalmology, University of Pittsburgh School of medicine, Pittsburgh, PA 15213, United StatesDepartment of Ophthalmology, University of Pittsburgh School of medicine, Pittsburgh, PA 15213, United States; McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA 15213, United StatesDepartment of Ophthalmology, University of Pittsburgh School of medicine, Pittsburgh, PA 15213, United States; McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA 15213, United StatesDepartment of Ophthalmology, University of Pittsburgh School of medicine, Pittsburgh, PA 15213, United States; McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA 15213, United StatesDepartment of Ophthalmology, University of Pittsburgh School of medicine, Pittsburgh, PA 15213, United StatesIntroduction: Corneal blindness due to scarring is treated with corneal transplantation. However, a global problem is the donor material shortage. Preclinical and clinical studies have shown that cell-based therapy using corneal stromal stem cells (CSSCs) suppresses corneal scarring, potentially mediated by specific microRNAs transported in extracellular vesicles (EVs). However, not every CSSC batch from donors achieves similar anti-scarring effects. Objectives: To examine miRNA profiles in EVs from human CSSCs showing “healing” versus “non-healing” effects on corneal scarring and to design a tool to select CSSCs with strong healing potency for clinical applications. Methods: Small RNAs from CSSC-EVs were extracted for Nanostring nCounter Human miRNA v3 assay. MicroRNAs expressed > 20 folds in “healing” EVs (P < 0.05) were subject to enriched gene ontology (GO) term analysis. MiRNA groups with predictive regulation on inflammatory and fibrotic signalling were studied by mimic transfection to (1) mouse macrophages (RAW264.7) for M1 phenotype assay; (2) human corneal keratocytes for cytokine-induced fibrosis, and (3) human CSSCs for corneal scar prevention in vivo. The expression of miR-29a was screened in additional CSSC batches and the anti-scarring effect of cells was validated in mouse corneal wounds. Results: Twenty-one miRNAs were significantly expressed in “healing” CSSC-EVs and 9 miRNA groups were predicted to associate with inflammatory and fibrotic responses, and tissue regeneration (P <10−6). Overexpression of miR-29a and 381-5p significantly prevented M1 phenotype transition in RAW264.7 cells after lipopolysaccharide treatment, suppressed transforming growth factor β1-induced fibrosis marker expression in keratocytes, and reduced scarring after corneal injury. High miR-29a expression in EV fractions distinguished human CSSCs with strong healing potency, which inhibited corneal scarring in vivo. Conclusion: We characterized the anti-inflammatory and fibrotic roles of miR-29a and 381-5p in CSSCs, contributing to scar prevention. MiR-29a expression in EVs distinguished CSSCs with anti-scarring quality, identifying good quality cells for a scarless corneal healing.http://www.sciencedirect.com/science/article/pii/S2090123222001205Corneal blindnessScarringStromal cell therapyStem cellsExtracellular vesiclesmiR-29a |
| spellingShingle | Gary Hin-Fai Yam Tianbing Yang Moira L Geary Mithun Santra Martha Funderburgh Elizabeth Rubin Yiqin Du Jose A Sahel Vishal Jhanji James L Funderburgh Human corneal stromal stem cells express anti-fibrotic microRNA-29a and 381-5p – A robust cell selection tool for stem cell therapy of corneal scarring Journal of Advanced Research Corneal blindness Scarring Stromal cell therapy Stem cells Extracellular vesicles miR-29a |
| title | Human corneal stromal stem cells express anti-fibrotic microRNA-29a and 381-5p – A robust cell selection tool for stem cell therapy of corneal scarring |
| title_full | Human corneal stromal stem cells express anti-fibrotic microRNA-29a and 381-5p – A robust cell selection tool for stem cell therapy of corneal scarring |
| title_fullStr | Human corneal stromal stem cells express anti-fibrotic microRNA-29a and 381-5p – A robust cell selection tool for stem cell therapy of corneal scarring |
| title_full_unstemmed | Human corneal stromal stem cells express anti-fibrotic microRNA-29a and 381-5p – A robust cell selection tool for stem cell therapy of corneal scarring |
| title_short | Human corneal stromal stem cells express anti-fibrotic microRNA-29a and 381-5p – A robust cell selection tool for stem cell therapy of corneal scarring |
| title_sort | human corneal stromal stem cells express anti fibrotic microrna 29a and 381 5p a robust cell selection tool for stem cell therapy of corneal scarring |
| topic | Corneal blindness Scarring Stromal cell therapy Stem cells Extracellular vesicles miR-29a |
| url | http://www.sciencedirect.com/science/article/pii/S2090123222001205 |
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