Saori Yaguchi

Angiotensin II Type 1 Receptor Antagonist Attenuates Lacrimal Gland, Lung, and Liver Fibrosis in a Murine Model of Chronic Graft-Versus-Host Disease

Chronic graft-versus-host disease (cGVHD), a serious complication following allogeneic HSCT (hematopoietic stem cell transplantation), is characterized by systemic fibrosis. The tissue renin-angiotensin system (RAS) is involved in the fibrotic pathogenesis, and an angiotensin II type 1 receptor (AT1R) antagonist can attenuate fibrosis. Tissue RAS is present in the lacrimal gland, lung, and liver, and is known to be involved in the fibrotic pathogenesis of the lung and liver. This study aimed to determine whether RAS is involved in fibrotic pathogenesis in the lacrimal gland and to assess the effect of an AT1R antagonist on preventing lacrimal gland, lung, and liver fibrosis in cGVHD model mice. We used the B10.D2→BALB/c (H-2d) MHC-compatible, multiple minor histocompatibility antigen-mismatched model, which reflects clinical and pathological symptoms of human cGVHD. First, we examined the localization and expression of RAS components in the lacrimal glands using immunohistochemistry and quantitative real-time polymerase chain reaction (PCR). Next, we administered an AT1R antagonist (valsartan; 10 mg/kg) or angiotensin II type 2 receptor (AT2R) antagonist (PD123319; 10 mg/kg) intraperitoneally into cGVHD model mice and assessed the fibrotic change in the lacrimal gland, lung, and liver. We demonstrated that fibroblasts expressed angiotensin II, AT1R, and AT2R, and that the mRNA expression of angiotensinogen was greater in the lacrimal glands of cGVHD model mice than in controls generated by syngeneic-HSCT. The inhibition experiment revealed that fibrosis of the lacrimal gland, lung, and liver was suppressed in mice treated with the AT1R antagonist, but not the AT2R antagonist. We conclude that RAS is involved in fibrotic pathogenesis in the lacrimal gland and that AT1R antagonist has a therapeutic effect on lacrimal gland, lung, and liver fibrosis in cGVHD model mice. Our findings point to AT1R antagonist as a possible target for therapeutic intervention in cGVHD.

Surgical management of lacrimal punctal cauterization in chronic GVHD-related dry eye with recurrent punctal plug extrusion

We investigated the efficacy of lacrimal punctal occlusion surgery with a cautery device in patients with chronic GVHD (cGVHD)-related dry eye, with recanalization of puncta and recurrent punctal plug extrusion. A total of 23 puncta from 14 eyes of 10 patients with chronic GVHD (cGVHD)-related dry eye underwent punctual thermal cauterization with a high-temperature disposable cautery device. All patients were refractory to conventional treatment, including artificial tear eye drops, autologous serum eye drops and vitamin A eye drops, and had a history of recanalization and recurrent punctal plug extrusion. The effect of lacrimal punctal cauterization by thermal cautery device was evaluated by changes in subjective symptom scores, corrected distance visual acuity, Schirmers test values, fluorescein staining scores, rose bengal staining scores, and tear-film break-up time before and 3 months after the surgery. Subjective symptom scores, Schirmers test values, fluorescein and rose bengal scores, and tear-film break-up time improved significantly 3 months after the surgery. Recanalization of puncta was not observed in all the cases (0 of 14 eyes, 0%). Lacrimal punctal cauterization was effective with no recanalization and significant improvements in subjective symptoms and the ocular surface environment in cGVHD-related dry eye patients who had been refractory to conventional treatments.

Mucosal microvilli in dry eye patients with chronic GVHD

The ocular surface is a frequent target tissue of mucosal chronic GVHD (cGVHD). We investigated the histopathological features of the conjunctival microvilli in patients with cGVHD. Conjunctival tissue specimens from patients with cGVHD or Sjögrens syndrome (SS) or from healthy individuals were examined by light microscopy and EM, impression cytology, and immunohistochemistry. The cGVHD conjunctivae showed significantly more metaplasia and fewer goblet cells than the SS and normal conjunctivae. Abundant CD8+ T cells infiltrated the basal epithelia in the cGVHD conjunctiva. The microvilli per standard epithelial unit and the secretory vesicles were counted by analyzing electron micrographs. The mean number of mucosal microvilli was significantly lower in the cGVHD than that in the SS or normal specimens, and the microvilli were significantly shorter, with a smaller height–width ratio. The mean number of secretory vesicles was also significantly lower, and the membrane-spanning mucin thinner, in the cGVHD compared with the SS and normal specimens. Thus, the conjunctival mucosal microvilli of cGVHD patients were significantly different in number and morphology from those of SS and normal subjects. These may be important factors affecting the stability of the tear-film layer and its contribution to cGVHD-related dry eye.

Presence and physiologic function of the renin-angiotensin system in mouse lacrimal gland

PURPOSE To investigate the expression, localization, and physiologic function of renin-angiotensin system (RAS) components in the mouse lacrimal gland. METHODS Lacrimal glands and cultured lacrimal gland fibroblasts from wild-type (WT) BALB/c (H-2(d)) mice were used. Reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry were used to determine the expression and localization of the RAS components, prorenin/renin, angiotensin-converting enzyme (ACE), angiotensin II, angiotensin II type 1 receptor (AT1R), and angiotensin II type 2 receptor (AT2R) in the normal mouse lacrimal gland. To examine the change in tear secretion, mice received ARB (AT1R blocker) or AT2R antagonist. Tear secretion was assessed by cotton thread test before and after drug administration. RESULTS The mRNAs coding for angiotensinogen, prorenin, ACE, and both AT1R and AT2R were found in normal lacrimal gland tissue and cultured lacrimal gland fibroblasts. Prorenin/renin and ACE were identified in myoepithelial cells around ducts and acini and in blood vessels. Angiotensin II, AT1R, and AT2R were observed in the ducts and interstitial fibroblasts. AT1R and AT2R were also localized in blood vessels. All the cultured lacrimal gland fibroblasts expressed angiotensin II, AT1R, and AT2R. Tear secretion increased in mice that received ARB. CONCLUSIONS The results are consistent with the hypothesis that a tissue-specific RAS is present in the lacrimal gland, and suggest that fibroblasts are one of the cell types playing a role in the tissue RAS. Tissue RAS might be involved in tissue function of regulating tear secretion in the lacrimal gland.

Foldable acrylic intraocular lens with distended haptics for transscleral fixation

&NA; We describe a foldable acrylic intraocular lens (IOL) with distended haptics suitable for transscleral fixation and the insertion procedure. The IOL has an acrylic optic and poly(methyl methacrylate) haptics with a microscopic indentation 1.3 mm from the tip. Transscleral fixation of the IOL was performed through corneal incisions in 22 eyes, and surgical results were retrospectively assessed. The IOL was sutured firmly in position using the cow‐hitch procedure, and there was no suture loosening to the distended haptic. The IOL design provided suitable fixation and may be indicated for bag fixation as well as transscleral fixation.

MHC-compatible bone marrow stromal/stem cells trigger fibrosis by activating host T cells in a scleroderma mouse model

Fibrosis of organs is observed in systemic autoimmune disease. Using a scleroderma mouse, we show that transplantation of MHC compatible, minor antigen mismatched bone marrow stromal/stem cells (BMSCs) play a role in the pathogenesis of fibrosis. Removal of donor BMSCs rescued mice from disease. Freshly isolated PDGFRα+ Sca-1+ BMSCs expressed MHC class II following transplantation and activated host T cells. A decrease in FOXP3+ CD25+ Treg population was observed. T cells proliferated and secreted IL-6 when stimulated with mismatched BMSCs in vitro. Donor T cells were not involved in fibrosis because transplanting T cell-deficient RAG2 knock out mice bone marrow still caused disease. Once initially triggered by mismatched BMSCs, the autoimmune phenotype was not donor BMSC dependent as the phenotype was observed after effector T cells were adoptively transferred into naïve syngeneic mice. Our data suggest that minor antigen mismatched BMSCs trigger systemic fibrosis in this autoimmune scleroderma model. DOI: http://dx.doi.org/10.7554/eLife.09394.001

Long-term rebamipide and diquafosol in two cases of immune-mediated dry eye.

Purpose Two new drugs with mucin-inducing and secretion-promotive effects, rebamipide and diquafosol, were recently approved as topical dry-eye treatments. We report two cases in which the long-term use of mucin-inducing eye drops improved chronic ocular graft-versus-host disease (cGVHD)–related dry eye and ocular cicatricial pemphigoid (OCP)-like disease. Case Reports Case 1. A 61-year-old woman had cGVHD-related dry eye that resisted traditional medications. Next, we use topical diquafosol in addition to conventional treatments. The patient used diquafosol for 6 months without experiencing any side effects. The symptoms, including dry-eye sensation, ocular pain, foreign body sensation, and photophobia, as well as ocular surface findings including fluorescein and rose bengal scores and tear break-up time (TBUT), partly improved. To further improve the clinical signs and symptoms and decrease chronic inflammation, rebamipide was added to diquafosol. The symptoms, TBUT, and fluorescein and rose bengal scores markedly improved after long-term dual treatment without any side effects for 6 months. Case 2. A 77-year-old woman had OCP-like disease with dry eye. The patient did not improve using the currently available conventional treatments. Next, we use topical rebamipide in addition to conventional treatments. Symptoms including asthenopia, dry-eye sensation, ocular pain, and dull sensation, as well as fluorescein and rose bengal scores and TBUT, partly improved. Specifically, functional visual acuity was markedly improved after commencement of rebamipide. To further improve the clinical signs and symptoms and increase tear film stability and tear film volume, diquafosol was added to rebamipide. The combination of diquafosol and rebamipide worked for the patient. Improvements were seen in several symptoms, fluorescein and rose bengal scores, Schirmer test value, and TBUT without any side effects for 12 months. Conclusions Long-term treatment with topical rebamipide and diquafosol can improve dry eye in patients with cGVHD or OCP-like disease.

Objective Evaluation of Zonular Weakness: Measurement of Lens Movement atthe Start of Capsulorhexis Using Extracted Porcine Eyes

Objective: To evaluate the degree of zonular weakness based on lens movement at the start of capsulorhexis using extracted porcine eyes. Methods: Zonular dehiscence of 18°, 36°, and 54° was created and alternated with healthy areas into five zones to create a model simulation of weakness of the zonule of Zinn corresponding to dehiscence of 90°, 180°, and 270°. During continuous curvilinear capsulorhexis (CCC), an image displaying the anterior capsule grasped by a cystotome and another displaying the initial tear were captured, and the two images were superimposed to measure the movement distance of the cortical opacities created using a hook. Porcine eyes with no dehiscence were used as the control group. Results: Compared with a distance of 0.44 ± 0.13 mm in the control group, the cortical opacity movement distances in the weakness group with dehiscence corresponding to 90°, 180°, and 270° were 0.68 ± 0.27, 1.01 ± 0.22, and 1.32 ± 0.35 mm, respectively. Significant differences were observed between the control, 90°, 180°, and 270° dehiscence groups (P<0.001). Conclusion: This study revealed numerical data for the degree of weakness of the zonule of Zinn in porcine eyes. Clinical application of this method may be useful for establishing surgical procedures and prognoses for cataract surgery in patients with weakness of the zonule of Zinn.

Categorization and Surgical Techniques of Weak Zonule Based on Findings at Capsulorhexis during Cataract Surgery

Purpose: We categorize weak zonule during capsulorhexis. Design: Retrospective interventional case series. Methods: The study examined 5447 consecutive eyes of 3527 cases that underwent phacoemulsification (PEA) and intraocular lens (IOL) implantation between March 2006 and March 2014 at Showa University Fujigaoka Hospital, Kanagawa, Japan. Weak zonule was categorized based on findings at capsulorhexis and difficulty in performing continuous curvilinear capsulorhexis (CCC). The categorization definitions were as follows: (1) Group N (normal) with no or slight lens movement at the start of the CCC and no difficulty in performing CCC, (2) Group W (weak) with moderate lens movement frequently accompanied by fold formation during CCC and some difficulty in performing CCC, (3) Group VW (very weak) because of severe lens movement and difficulties with the initial puncture, CCC can be performed with the aid of a highly retentive and cohesive ophthalmic viscoelastic device (OVD; Healon 5®), and (4) Group EW (extremely weak) with zonular deficiency in addition to the Group VW criteria and accounted for the cases of severe phacodonesis, lens subluxation, lens luxation into the anterior chamber, and dropped nucleus into the vitreous cavity that could be categorized preoperatively. We examined capsule stabilization device use during PEA, surgical lens removal, and IOL fixation in each group. Results: We defined 5098 eyes as Group N, 251 eyes as Group W, 55 eyes as Group VW, and 43 eyes as Group EW. As the zonule became weak, the use of the capsule stabilization device increased in PEA and intracapsular cataract extraction, and scleral suture fixation of IOL increased. Pars plana vitrectomy for lens removal was performed in 5 eyes (11.63%) categorized as Group EW. Conclusions: The categorization of weak zonule at CCC may be useful for selecting an appropriate capsule stabilization device and procedure during cataract surgery.

Tissue Renin-Angiotensin System in Lacrimal Gland Fibrosis in a Murine Model of Chronic Graft-Versus-Host Disease.

Abstract: Chronic graft-versus-host disease (cGVHD) is a serious complication known to occur after allogeneic hematopoietic stem cell transplantation. Clinical manifestation includes inflammation and fibrosis. Many peripheral tissues are capable of generating the renin–angiotensin system (RAS) components, called the tissue RAS, and have various roles in tissue-specific physiological and pathological functions of inflammation and fibrosis. This article reviews evidence for the presence of the tissue RAS in the normal mouse lacrimal gland, the role of the tissue RAS in the fibrotic pathogenesis of the lacrimal gland in cGVHD model mice, and the effect of angiotensin II receptor blockers on preventing lacrimal gland fibrosis. B10.D2→BALB/c (H-2d) major histocompatibility complex-compatible, minor histocompatibility antigen-mismatched mice were used as a model of cGVHD, which reflects the clinical and pathological symptoms of human cGVHD. We also describe the localization of RAS components in the normal mouse lacrimal gland. In addition, we characterize the inflammatory and fibrotic changes of the lacrimal gland in cGVHD model mice, demonstrate that fibroblasts strongly express angiotensin II, angiotensin II type 1 receptor (AT1R), and angiotensin II type 2 receptor, and show that mRNA expression of angiotensinogen increased in the lacrimal gland of cGVHD model mice. Inhibitory experiments revealed that lacrimal gland fibrosis was suppressed in mice treated with an AT1R blocker, but not in mice treated with an angiotensin II type 2 receptor blocker. Hence, we conclude that the tissue RAS is involved in the fibrotic pathogenesis of the lacrimal gland and that AT1R blockers have a therapeutic effect on lacrimal gland fibrosis in cGVHD model mice.