COL8A2 Regulates the Fate of Corneal Endothelial Cells

Hwang, Jiyoung; Joong, Dae; Choi, Jinju; Shin, Young Joo

doi:10.1167/iovs.61.11.26

Cited by 24 publications

(32 citation statements)

References 54 publications

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“…Differently, COL8A2 expression significantly increased at day 14. The EnMT-like changes in the cultured human CECs could be attenuated by inhibiting the expression of COL8A2 [ 47 ]. The increase in COL8A2 observed here might be associated with the EnMT process of the transplanted cells observed here at the later stage.…”

Section: Discussionmentioning

confidence: 99%

Transplantation of human induced pluripotent stem cell-derived neural crest cells for corneal endothelial regeneration

et al. 2021

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Background The corneal endothelium maintains corneal hydration through the barrier and pump function, while its dysfunction may cause corneal edema and vision reduction. Considering its development from neural crest cells (NCCs), here we investigated the efficacy of the human induced pluripotent stem cell (hiPSC)-derived NCCs for corneal endothelial regeneration in rabbits. Methods Directed differentiation of hiPSC-derived NCCs was achieved using the chemically defined medium containing GSK-3 inhibitor and TGF-β inhibitor. The differentiated cells were characterized by immunofluorescence staining, FACS analysis, and in vitro multi-lineage differentiation capacity. For in vivo functional evaluation, 1.0 × 106 hiPSC-derived NCCs or NIH-3 T3 fibroblasts (as control) combined with 100 μM Y-27632 were intracamerally injected into the anterior chamber of rabbits following removal of corneal endothelium. Rabbit corneal thickness and phenotype changes of the transplanted cells were examined at 7 and 14 days with handy pachymeter, dual-immunofluorescence staining, and quantitative RT-PCR. Results The hiPSC-derived NCCs were differentiated homogenously through 7 days of induction and exhibited multi-lineage differentiation capacity into peripheral neurons, mesenchymal stem cells, and corneal keratocytes. After 7 days of intracameral injection in rabbit, the hiPSC-derived NCCs led to a gradual recovery of normal corneal thickness and clarity, when comparing to control rabbit with fibroblasts injection. However, the recovery efficacy after 14 days deteriorated and caused the reappearance of corneal edema. Mechanistically, the transplanted cells exhibited the impaired maturation, cellular senescence, and endothelial-mesenchymal transition (EnMT) after the early stage of the in vivo directional differentiation. Conclusions Transplantation of the hiPSC-derived NCCs rapidly restored rabbit corneal thickness and clarity. However, the long-term recovery efficacy was impaired by the improper maturation, senescence, and EnMT of the transplanted cells.

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Section: Discussionmentioning

confidence: 99%

Transplantation of human induced pluripotent stem cell-derived neural crest cells for corneal endothelial regeneration

et al. 2021

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“…This study was reviewed and approved by the institutional review board/ethics committee of Hallym University Kangnam Sacred Heart Hospital and was conducted in accordance with the Helsinki Declaration. Cells were cultured in accordance with previously published methods [ 72 ]. The corneas from six donors were used.…”

Section: Methodsmentioning

confidence: 99%

“…hCECs were detached from Descemet’s membrane by trypsinizing for 10 min. The cells were cultured in 6-well plates applied with a fibronectin–collagen combination (FNC) coating mix (Athena Environmental Sciences, Inc., Baltimore, MD) and passaged at a ratio of 1:3 [ 72 ].…”

Section: Methodsmentioning

confidence: 99%

“…After incubation with CCK-8 solution for 1–2 h, a Synergy HTX (BioTEK, Winooski, VT) multi-mode reader was used for evaluating cell viability by measuring optical density (OD) at 450 nm [ 73 ]. A bromodeoxyuridine (BrdU) incorporation assay kit (Roche Diagnostics, GmbH, Mannheim, Germany) was employed for evaluating cell proliferation rate according to manufacturer’s protocol [ 72 ].…”

Section: Methodsmentioning

confidence: 99%

“…After washing, an HRP-conjugated secondary antibody and a WEST-Queen™ western blot detection kit (iNtRON Biotechnology, Seongnam, Kyounggi-do) were applied for the detection of protein bands. Video image analysis (Luminograph II, Atto, Tokyo) was used to quantify the data [ 72 ].…”

Section: Methodsmentioning

confidence: 99%

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miR-30c-1 encourages human corneal endothelial cells to regenerate through ameliorating senescence

Younghwan

Hwang

Shin

2021

Aging

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In the present study, we studied the role of microRNA-30c-1 (miR-30c-1) on transforming growth factor beta1 (TGF-β1)-induced senescence of hCECs. hCECs were transfected by miR-30c-1 and treated with TGF-β1 to assess the inhibitory effect of miR-30c-1 on TGF-β1-induced senescence. Cell viability and proliferation rate in miR-30c-1-transfected cells was elevated compared with control. Cell cycle analysis revealed that cell abundance in S phase was elevated in miR-30c-1-treated cells compared with control. TGF-β1 increased the senescence of hCECs; however, this was ameliorated by miR-30c-1. TGF-β1 increased the size of hCECs, the ratio of senescence-associated beta-galactosidase-stained cells, secretion of senescence-associated secretory phenotype factors, the oxidative stress, and arrested the cell cycle, all of which were ameliorated by miR-30c-1 treatment. miR-30c-1 also suppressed a TGF-β1-induced depolarization of mitochondrial membrane potential and a TGF-β1 stimulated increase in levels of cleaved poly (ADP-ribose) polymerase (PARP), cleaved caspase 3, and microtubule-associated proteins 1A/1B light chain 3B II. In conclusion, miR-30c-1 promoted the proliferation of hCECs through ameliorating the TGF- β1-induced senescence of hCECs and reducing cell death of hCECs. Thus, miR-30c-1 may be a therapeutic target for hCECs regeneration.

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Nanoneedles Induce Targeted siRNA Silencing of p16 in the Human Corneal Endothelium

et al. 2022

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Nanoneedles can target nucleic acid transfection to primary cells at tissue interfaces with high efficiency and minimal perturbation. The corneal endothelium is an ideal target for nanoneedle‐mediated RNA interference therapy aimed at enhancing its proliferative capacity, necessary for tissue regeneration. This work develops a strategy for siRNA nanoninjection to the human corneal endothelium. Nanoneedles can deliver p16‐targeting siRNA to primary human corneal endothelial cells in vitro without toxicity. The nanoinjection of siRNA induces p16 silencing and increases cell proliferation, as monitored by ki67 expression. Furthermore, siRNA nanoinjection targeting the human corneal endothelium is nontoxic ex vivo, and silences p16 in transfected cells. These data indicate that nanoinjection can support targeted RNA interference therapy for the treatment of endothelial corneal dysfunction.

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COL8A2 Regulates the Fate of Corneal Endothelial Cells

Cited by 24 publications

References 54 publications

Transplantation of human induced pluripotent stem cell-derived neural crest cells for corneal endothelial regeneration

Transplantation of human induced pluripotent stem cell-derived neural crest cells for corneal endothelial regeneration

miR-30c-1 encourages human corneal endothelial cells to regenerate through ameliorating senescence

Nanoneedles Induce Targeted siRNA Silencing of p16 in the Human Corneal Endothelium

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