Agrin Promotes Limbal Stem Cell Proliferation and Corneal Wound Healing Through Hippo-Yap Signaling Pathway

Hou, Lulu; Fu, Wei; Liu, Ying; Wang, Qun; Wang, Liqiang; Huang, Yifei

doi:10.1167/iovs.61.5.7

Cited by 20 publications

(13 citation statements)

References 42 publications

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“…Previous studies on decellularized limbal tissue provided little detailed examinations of ECM components [ 20 , 21 ]. Agrin, a heparin sulfate proteoglycan that promotes LEPC proliferation and corneal wound healing [ 41 ], is highly expressed in the limbal region and its expression declines towards the central cornea [ 25 ]. Agrin was preserved in the DHL scaffolds of the current study.…”

Section: Discussionmentioning

confidence: 99%

A Decellularized Human Limbal Scaffold for Limbal Stem Cell Niche Reconstruction

Polisetti

Roschinski

Schlötzer‐Schrehardt

et al. 2021

IJMS

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The transplantation of ex vivo expanded limbal epithelial progenitor cells (LEPCs) on amniotic membrane or fibrin gel is an established therapeutic strategy to regenerate the damaged corneal surface in patients with limbal stem cell deficiency (LSCD), but the long-term success rate is restricted. A scaffold with niche-specific structure and extracellular matrix (ECM) composition might have the advantage to improve long-term clinical outcomes, in particular for patients with severe damage or complete loss of the limbal niche tissue structure. Therefore, we evaluated the decellularized human limbus (DHL) as a biomimetic scaffold for the transplantation of LEPCs. Corneoscleral tissue was decellularized by sodium deoxycholate and deoxyribonuclease I in the presence or absence of dextran. We evaluated the efficiency of decellularization and its effects on the ultrastructure and ECM composition of the human corneal limbus. The recellularization of these scaffolds was studied by plating cultured LEPCs and limbal melanocytes (LMs) or by allowing cells to migrate from the host tissue following a lamellar transplantation ex vivo. Our decellularization protocol rapidly and effectively removed cellular and nuclear material while preserving the native ECM composition. In vitro recellularization by LEPCs and LMs demonstrated the good biocompatibility of the DHL and intrastromal invasion of LEPCs. Ex vivo transplantation of DHL revealed complete epithelialization as well as melanocytic and stromal repopulation from the host tissue. Thus, the generated DHL scaffold could be a promising biological material as a carrier for the transplantation of LEPCs to treat LSCD.

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Section: Discussionmentioning

confidence: 99%

A Decellularized Human Limbal Scaffold for Limbal Stem Cell Niche Reconstruction

Polisetti

Roschinski

Schlötzer‐Schrehardt

et al. 2021

IJMS

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“…In the ocular surface, agrin is highly expressed in the limbal regions partially colocalized with limbal stem cells (ABCG2 and p63-positive cells), and the expression of agrin decreases toward the conjunctiva (weakly expressed) and central cornea (absent or weakly expressed) ( Figure 4 ) ( Schlötzer-Schrehardt et al, 2007 ). A recent report has shown that agrin promotes limbal stem cell proliferation and corneal wound healing via the Hippo-Yap signaling pathway ( Hou et al, 2020 ). Conditional knockout of agrin in the cornea could potentially help further our understanding of the role of agrin in supporting limbal stem cells.…”

Section: Heparan Sulfate Proteoglycans (Hspgs)mentioning

confidence: 99%

Distribution and Function of Glycosaminoglycans and Proteoglycans in the Development, Homeostasis and Pathology of the Ocular Surface

Puri

Coulson-Thomas

Gesteira

et al. 2020

Front. Cell Dev. Biol.

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The ocular surface, which forms the interface between the eye and the external environment, includes the cornea, corneoscleral limbus, the conjunctiva and the accessory glands that produce the tear film. Glycosaminoglycans (GAGs) and proteoglycans (PGs) have been shown to play important roles in the development, hemostasis and pathology of the ocular surface. Herein we review the current literature related to the distribution and function of GAGs and PGs within the ocular surface, with focus on the cornea. The unique organization of ECM components within the cornea is essential for the maintenance of corneal transparency and function. Many studies have described the importance of GAGs within the epithelial and stromal compartment, while very few studies have analyzed the ECM of the endothelial layer. Importantly, GAGs have been shown to be essential for maintaining corneal homeostasis, epithelial cell differentiation and wound healing, and, more recently, a role has been suggested for the ECM in regulating limbal stem cells, corneal innervation, corneal inflammation, corneal angiogenesis and lymphangiogenesis. Reports have also associated genetic defects of the ECM to corneal pathologies. Thus, we also highlight the role of different GAGs and PGs in ocular surface homeostasis, as well as in pathology.

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“…DIANA-TarBase and microT-CDS algorithms (genes union function) were used to identify experimentally supported and predicted miRNA/target-mRNA interactions, respectively, as well as significant KEGG pathways, in which target genes were involved ( Table 2 and Table 3 ). Among them emerged the ECM-receptor interaction and the focal adhesion pathways, already described in cornea homeostasis and normal physiological functions [ 27 ], corneal epithelial cell motility [ 28 ], neovascularization [ 29 ] and adhesion [ 30 , 31 ], the Hippo signaling pathway, playing a role in limbal cell proliferation, corneal wound healing and regeneration [ 32 , 33 ] and, as expected, the Neurotrophin signaling pathway. The latter was considered for more in-depth analysis: sixty and fifty genes, targeted by 18 and 16 out of 21 miRNAs (miR-337-5p, mir-34a-3p, mir-1227-3p did not show any target gene) were detected by DIANA-TarBase and microT-based analysis, respectively ( Table 4 ).…”

Section: Resultsmentioning

confidence: 99%

MicroRNAs Expression in Response to rhNGF in Epithelial Corneal Cells: Focus on Neurotrophin Signaling Pathway

Compagnoni

Zelli

Bianchi

et al. 2022

IJMS

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Purpose. Nerve growth factor efficacy was demonstrated for corneal lesions treatment, and recombinant human NGF (rhNGF) was approved for neurotrophic keratitis therapy. However, NGF-induced molecular responses in cornea are still largely unknown. We analyzed microRNAs expression in human epithelial corneal cells after time-dependent rhNGF treatment. Methods. Nearly 700 microRNAs were analyzed by qRT-PCR. MicroRNAs showing significant expression differences were examined by DIANA-miRpath v.3.0 to identify target genes and pathways. Immunoblots were performed to preliminarily assess the strength of the in silico results. Results. Twenty-one microRNAs (miR-26a-1-3p, miR-30d-3p, miR-27b-5p, miR-146a-5p, miR-362-5p, mir-550a-5p, mir-34a-3p, mir-1227-3p, mir-27a-5p, mir-222-5p, mir-151a-5p, miR-449a, let7c-5p, miR-337-5p, mir-29b-3p, miR-200b-3p, miR-141-3p, miR-671-3p, miR-324-5p, mir-411-3p, and mir-425-3p) were significantly regulated in response to rhNGF. In silico analysis evidenced interesting target genes and pathways, including that of neurotrophin, when analyzed in depth. Almost 80 unique target genes (e.g., PI3K, AKT, MAPK, KRAS, BRAF, RhoA, Cdc42, Rac1, Bax, Bcl2, FasL) were identified as being among those most involved in neurotrophin signaling and in controlling cell proliferation, growth, and apoptosis. AKT and RhoA immunoblots demonstrated congruence with microRNA expression, providing preliminary validation of in silico data. Conclusions. MicroRNA levels in response to rhNGF were for the first time analyzed in corneal cells. Novel insights about microRNAs, target genes, pathways modulation, and possible biological responses were provided. Importantly, given the putative role of microRNAs as biomarkers or therapeutic targets, our results make available data which might be potentially exploitable for clinical applications.

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Agrin Promotes Limbal Stem Cell Proliferation and Corneal Wound Healing Through Hippo-Yap Signaling Pathway

Cited by 20 publications

References 42 publications

A Decellularized Human Limbal Scaffold for Limbal Stem Cell Niche Reconstruction

A Decellularized Human Limbal Scaffold for Limbal Stem Cell Niche Reconstruction

Distribution and Function of Glycosaminoglycans and Proteoglycans in the Development, Homeostasis and Pathology of the Ocular Surface

MicroRNAs Expression in Response to rhNGF in Epithelial Corneal Cells: Focus on Neurotrophin Signaling Pathway

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