Graft Versus Host Disease-Associated Dry Eye: Role of Ocular Surface Mucins and the Effect of Rebamipide, a Mucin Secretagogue

Abstract

PURPOSE. The present study was designed to investigate the role of ocular surface glycocalyx and mucins in graft versus host disease (GVHD)-associated dry eye. The ameliorative effect of topical rebamipide, a mucin secretagogue, on GVHD-associated dry eye was also tested. METHODS. A mouse model of allogeneic transplantation was used to induce ocular GVHD with C57BL/6 as donors and B6D2F1 as recipient mice. Phenol red thread method and fluorescein staining was used to quantify tear secretion and corneal keratopathy. At 8 weeks after the allogeneic transplantation, corneas were harvested to perform glycocalyx staining and confocal microscopy. Goblet cell staining was performed using periodic acid Schiff's staining. Corneal and tear film levels of Mucin 1, 4, 16, 19, and 5AC were quantified using ELISA and real-time PCR. Rebamipide was applied topically twice daily to mice eyes. RESULTS. Allogeneic transplantation resulted in ocular GVHD-associated dry eye characterized by a significant decrease in tear film volume and the onset of corneal keratopathy. Ocular GVHD caused a significant decrease in the area and thickness of corneal glycocalyx. A significant decrease in the goblet cells was also noted. A significant decrease in mucin 4 and 5AC levels was also observed. Topical treatment with rebamipide partially attenuated ocular GVHD-mediated decrease in tear film volume and significantly reduced the severity of corneal keratopathy. CONCLUSIONS. Ocular GVHD has detrimental impact on ocular surface glycocalyx and mucins. Rebamipide, a mucin secretagogue, partially prevents ocular GVHD-associated decrease in tear film and reduces the severity of corneal keratopathy.

Local Renin-Angiotensin System Activation and Myofibroblast Formation in Graft Versus Host Disease–Associated Conjunctival Fibrosis

“…The femurs and spleen cells were harvested from the donor mice, as described in our earlier publication. 27 Recipient female BALB/c mice were exposed to 700-cGy irradiation using an x-ray irradiator (RS2000 Series Biological Irradiator; Rad Source Technologies, Suwanee, GA, USA). After 6 hours, the irradiated recipient female BALB/c mice were injected with 2 × 10 6 spleen cells and 1 × 10 6 bone marrow cells.…”

“…Tear volume was quantified before the allogeneic and syngeneic transplantation (baseline) and at weeks 2, 4, 6, and 8 after the transplantation. 27 …”

Chronic Ocular Graft Versus Host Disease: An Update and Review

“…171 Also, rebamipide, a mucin secretagogue, showed improved keratopathy and tear film in a mouse model of oGVHD. 172 A mouse model and limited human study examined whether a SNARE protein vesicle-associated membrane protein 8 (VAMP8) was associated with the development of chronic oGVHD. The expression of VAMP8 in the chronic GVHD affected population was decreased, causing decreased tear secretion changes.…”

Ocular Graft-versus-Host Disease in a Chemotherapy-Based Minor-Mismatch Mouse Model Features Corneal (Lymph-) Angiogenesis

“…Regarding the eye, several pre-clinical models are established to specifically investigate ocular manifestations of GVHD [17][18][19][20], as recently reviewed in [4]. In these models, several phenotypes similar to either acute or chronic ocular GVHD in humans are described and investigated further, including inflammation and fibrosis of the cornea, conjunctiva, lid marging, and lacrimal glands [18,19,[21][22][23][24]. Some of these models use GFP-overexpressing cells [21].…”