Murata Miyuki
| Faculty of Medicine Specialized Medicine Sensory Organ Medicine | Assistant Professor |
| Hokkaido University Hospital | Assistant Professor |
Last Updated :2025/06/07
■Researcher basic information
Researchmap personal page
Home Page URL
J-Global ID
Educational Organization
- Bachelor's degree program, Departments of Medicine, School of Medicine
- Master's degree program, Graduate School of Medicine
- Doctoral (PhD) degree program, Graduate School of Medicine
■Career
Career
- Apr. 2023 - Present
Hokkaido University, 大学院医学研究院眼科学教室, 助教, Japan - Apr. 2016 - Mar. 2023
北海道大学大学院医学研究院, 眼循環代謝学講座, 特任助教 - Sep. 2011 - Mar. 2014
Laboratory of Metabolism, National Cancer Institute, National Institutes of Health, United States - Apr. 2010 - Mar. 2011
Hokkaido University, 大学院医学研究院炎症眼科学講座, 特任助教, Japan - Oct. 2009 - Mar. 2010
Hokkaido University, 大学院医学研究院眼科学教室, 学術研究員, Japan - Apr. 2002 - Mar. 2007
Sapporo Medical University, 医学部生物学教室, 研究生, Japan - Jun. 1999 - Mar. 2002
Sapporo Medical University, 医学部内科学第一講座, 研究生, Japan
■Research activity information
Papers
- Clinicopathological findings in refractory diabetic macular edema: A case report.
Takayuki Tanaka, Satoru Kase, Michiyuki Saito, Ikuyo Hirose, Miyuki Murata, Emi Takakuwa, Susumu Ishida
Biomedical reports, 20, 1, 13, 13, Jan. 2024, [International Magazine]
English, The present study describes the case of a patient with refractory diabetic cystoid macular edema who underwent vitrectomy with en bloc removal of the cystoid lesion component. The current study also performed histopathological and immunohistochemical analyses of the cystoid lesion component to assess fibrin/fibrinogen and advanced glycation end-products (AGEs) immunoreactivity. A 69-year-old Japanese man presented with visual loss in the left eye due to residual cystoid macular edema (CME) refractory to anti-vascular endothelial growth factor therapy. Best-corrected visual acuity was 1.2 in the right eye (OD) and 0.5 in the left eye (OS). Fundus examination showed dot hemorrhages and hard exudates in the peri-macular region with pan-retinal photocoagulation scars in both eye. Swept-source optical coherence tomography revealed CME with slight hyperreflectivity in the cyst OS. A total of 3 months after the initial visit, pars plana vitrectomy was performed, and the translucent solidified component within the cystoid lesion was isolated. Histopathologically, the excised component was elliptical in shape, measuring 0.7x0.4 mm and exhibited homogeneous eosinophilic material without cellular components. No membranous structure was observed surrounding the component. Immunohistochemistry demonstrated that the tissue was positive for fibrin/fibrinogen and weakly positive for AGEs, but was negative for glial fibrillary acidic protein, type 1 collagen and receptor for AGEs. To the best of our knowledge, the present case report is the first to histopathologically examine the contents of refractory CME, and to immunohistochemically demonstrate that fibrin in diabetic CME may be post-translationally modified by AGEs. These results suggested that fibrin in CME may escape degradation by plasmin due to post-translational modifications. - Phosphorylation of αB-Crystallin Involves Interleukin-1β-Mediated Intracellular Retention in Retinal Müller Cells: A New Mechanism Underlying Fibrovascular Membrane Formation.
Taku Yamamoto, Satoru Kase, Akihiro Shinkai, Miyuki Murata, Kasumi Kikuchi, Di Wu, Yasushi Kageyama, Masami Shinohara, Tomohiko Sasase, Susumu Ishida
Investigative ophthalmology & visual science, 64, 10, 20, 20, 03 Jul. 2023, [International Magazine]
English, Scientific journal, PURPOSE: Chronic inflammation plays a pivotal role in the pathology of proliferative diabetic retinopathy (PDR), in which biological alterations of retinal glial cells are one of the key elements. The phosphorylation of αB-crystallin/CRYAB modulates its molecular dynamics and chaperone activity, and attenuates αB-crystallin secretion via exosomes. In this study, we investigated the effect of phosphorylated αB-crystallin in retinal Müller cells on diabetic mimicking conditions, including interleukin (IL)-1β stimuli. METHODS: Human retinal Müller cells (MIO-M1) were used to examine gene and protein expressions with real-time quantitative PCR, enzyme linked immunosorbent assay (ELISA), and immunoblot analyses. Cell apoptosis was assessed by Caspase-3/7 assay and TdT-mediated dUTP nick-end labeling staining. Retinal tissues isolated from the Spontaneously Diabetic Torii (SDT) fatty rat, a type 2 diabetic animal model with obesity, and fibrovascular membranes from patients with PDR were examined by double-staining immunofluorescence. RESULTS: CRYAB mRNA was downregulated in MIO-M1 cells with the addition of 10 ng/mL IL-1β; however, intracellular αB-crystallin protein levels were maintained. The αB-crystallin serine 59 (Ser59) residue was phosphorylated with IL-1β application in MIO-M1 cells. Cell apoptosis in MIO-M1 cells was induced by CRYAB knockdown. Immunoreactivity for Ser59-phosphorylated αB-crystallin and glial fibrillary acidic protein was colocalized in glial cells of SDT fatty rats and fibrovascular membranes. CONCLUSIONS: The Ser59 phosphorylation of αB-crystallin was modulated by IL-1β in Müller cells under diabetic mimicking inflammatory conditions, suggesting that αB-crystallin contributes to the pathogenesis of PDR through an anti-apoptotic effect. - Effects of Mirogabalin on Hyperalgesia and Chronic Ocular Pain in Tear-Deficient Dry-Eye Rats.
Kasumi Kikuchi, Yoshiaki Tagawa, Miyuki Murata, Susumu Ishida
Investigative ophthalmology & visual science, 64, 5, 27, 27, 01 May 2023, [International Magazine]
English, Scientific journal, PURPOSE: Patients with dry eye disease (DED) sometimes complain of ocular pain. DED-related ocular pain has many similarities with neuropathic pain. Mirogabalin, a novel ligand for the α2δ subunit of voltage-gated calcium channels, is approved for treating neuropathic pain in Japan. This study aimed to investigate the effect of mirogabalin on hyperalgesia and chronic ocular pain in a rat DED model. METHODS: DED was induced in female Sprague Dawley rats by unilaterally excising the external lacrimal gland (ELG) and Harderian gland (HG). After 4 weeks of ELG and HG removal, tear production (pH threads) and corneal epithelial damage (fluorescein staining) were evaluated. Corneal hyperalgesia and chronic pain were analyzed, respectively, by measuring capsaicin-induced eye-wiping behavior and c-Fos expression in the trigeminal nucleus. Mirogabalin (10 or 3 mg/kg) was evaluated for effects on DED-induced hyperalgesia and chronic ocular pain. RESULTS: Tear production was significantly lower in DED-induced eyes than in control eyes. Corneal damage was significantly higher in DED eyes than in control eyes. Hyperalgesia and chronic ocular pain were detected 4 weeks after ELG and HG removal. Five days of mirogabalin administration significantly suppressed capsaicin-induced eye-wiping behavior, which indicated the suppression of ocular hyperalgesia. Administration of 10 mg/kg mirogabalin significantly reduced c-Fos expression in the trigeminal nucleus, which indicated the amelioration of chronic ocular pain. CONCLUSIONS: Mirogabalin suppressed DED-induced hyperalgesia and chronic ocular pain in a rat DED model. Our findings suggested that mirogabalin might effectively alleviate chronic ocular pain in patients with DED. - Involvement of Angiopoietin 2 and vascular endothelial growth factor in uveitis.
Kayo Suzuki, Daiju Iwata, Kenichi Namba, Keitaro Hase, Miki Hiraoka, Miyuki Murata, Nobuyoshi Kitaichi, Richard Foxton, Susumu Ishida
PloS one, 18, 11, e0294745, 2023, [International Magazine]
English, Scientific journal, PURPOSE: Angiopoietin (Ang) 2 is released from vascular endothelial cells by the stimulation of vascular endothelial growth factor (VEGF)A. Ang2 increases the expression of leukocyte adhesion molecules on endothelial cells via nuclear factor κB. The aim of this study was to evaluate the effects of Ang2 and VEGFA on ocular autoimmune inflammation. METHODS: We measured the concentrations of Ang2 and VEGFA in vitreous samples among patients with uveitis. Vitreous samples were collected from 16 patients with idiopathic uveitis (uveitis group) and 16 patients with non-inflammatory eye disease (control group). Experimental autoimmune uveoretinitis (EAU) was induced in B10.BR mice with a human interphotoreceptor retinoid-binding protein-derived peptide. The retinochoroidal tissues of the EAU mice were removed, and the mRNA levels of Ang2 and VEGFA were examined. EAU mice treated with anti-Ang2, anti-VEGFA, a combination of anti-Ang2 and anti-VEGFA, anti-Ang2/VEGFA bispecific, or IgG control antibodies were clinically and histopathologically evaluated. RESULTS: The protein levels of Ang2 and VEGFA were significantly higher in the vitreous samples of patients with uveitis than in controls (P<0.05). The retinochoroidal mRNA levels of Ang2 and VEGFA were significantly upregulated in EAU mice compared to controls (n = 6, P<0.05). Although there was no significant difference, treatment with anti-VEGFA antibody reduced the clinical and histopathological scores. However, treatment with anti-Ang2 antibody reduced the clinical and histopathological scores (n = 18-20, P<0.05). Furthermore, these scores were further decreased when treated by inhibiting both Ang2 and VEGFA. CONCLUSIONS: Based on these results, VEGFA and Ang2 were shown to be upregulated locally in the eye of both uveitis patients and models of uveitis. Dual inhibition of Ang2 and VEGFA is suggested to be a new therapeutic strategy for uveitis. - Relationship between Brachial-Ankle Pulse Wave Velocity and Fundus Arteriolar Area Calculated Using a Deep-Learning Algorithm.
Kanae Fukutsu, Michiyuki Saito, Kousuke Noda, Miyuki Murata, Satoru Kase, Ryosuke Shiba, Naoki Isogai, Yoshikazu Asano, Nagisa Hanawa, Mitsuru Dohke, Manabu Kase, Susumu Ishida
Current eye research, 47, 11, 1534, 1537, Nov. 2022, [International Magazine]
English, Scientific journal, PURPOSE: Retinal vessels reflect alterations related to hypertension and arteriosclerosis in the physical status. Previously, we had reported a deep-learning algorithm for automatically detecting retinal vessels and measuring the total retinal vascular area in fundus photographs (VAFP). Herein, we investigated the relationship between VAFP and brachial-ankle pulse wave velocity (baPWV), which is the gold standard for arterial stiffness assessment in clinical practice. METHODS: Retinal photographs (n = 696) obtained from 372 individuals who visited the Keijinkai Maruyama Clinic for regular health checkups were used to analyze VAFP. Additionally, the baPWV was measured for each patient. Automatic retinal-vessel segmentation was performed using our deep-learning algorithm, and the total arteriolar area (AA) and total venular area (VA) were measured. Correlations between baPWV and several parameters, including AA and VA, were assessed. RESULTS: The baPWV was negatively correlated with AA (R = -0.40, n = 696, P < 2.2e-16) and VA (R = -0.36, n = 696, P < 2.2e-16). Independent variables (AA, sex, age, and systolic blood pressure) selected using the stepwise method showed a significant correlation with baPWV. The estimated baPWV, calculated using a regression equation with variables including AA, showed a better correlation with the measured baPWV (R = 0.70, n = 696, P < 2.2e-16) than the estimated value without AA (R = 0.68, n = 696, P < 2.2e-16). CONCLUSIONS: AA and VA were significantly correlated with baPWV. Moreover, baPWV estimated using AA correlated well with the actual baPWV. VAFP may serve as an alternative biomarker for evaluating systemic arterial stiffness. - Placental growth factor stabilizes VEGF receptor-2 protein in retinal pigment epithelial cells by downregulating glycogen synthase kinase 3 activity.
Miyuki Murata, Kousuke Noda, Satoru Kase, Keitaro Hase, Di Wu, Ryo Ando, Susumu Ishida
The Journal of biological chemistry, 298, 9, 102378, 102378, Sep. 2022, [International Magazine]
English, Scientific journal, Placental growth factor (PlGF) belongs to the vascular endothelial growth factor (VEGF) family of proteins that participate in angiogenesis and vasculogenesis. Anti-VEGF therapy has become the standard treatment for ocular angiogenic disorders in ophthalmological practice. However, there is emerging evidence that anti-VEGF treatment may increase the risk of atrophy of the retinal pigment epithelium (RPE), which is important for the homeostasis of retinal tissue. Whereas the cytoprotective role of VEGF family molecules, particularly that of VEGF A (VEGFA) through its receptor VEGF receptor-2 (VEGFR-2), has been recognized, the physiological role of PlGF in the retina is still unknown. In this study, we explored the role of PlGF in the RPE using PlGF-knockdown RPE cells generated by retrovirus-based PlGF-shRNA transduction. We show that VEGFA reduced apoptosis induced by serum starvation in RPE cells, whereas the antiapoptotic effect of VEGFA was abrogated by VEGFR-2 knockdown. Furthermore, PlGF knockdown increased serum starvation-induced cell apoptosis and unexpectedly reduced the protein level of VEGFR-2 in the RPE. The antiapoptotic effect of VEGFA was also diminished in PlGF-knockdown RPE cells. In addition, we found that glycogen synthase kinase 3 activity was involved in proteasomal degradation of VEGFR-2 in RPE cells and inactivated by PlGF via AKT phosphorylation. Overall, the present data demonstrate that PlGF is crucial for RPE cell viability and that PlGF supports VEGFA/VEGFR-2 signaling by stabilizing the VEGFR-2 protein levels through glycogen synthase kinase 3 inactivation. - Serum advanced glycation end-products and αB-crystallin in diabetic retinopathy patients.
Taku Yamamoto, Satoru Kase, Miyuki Murata, Susumu Ishida
Biomedical reports, 16, 4, 28, 28, Apr. 2022, [International Magazine]
English, Scientific journal, αB-crystallin, one of the small heat shock proteins, which is also known as HSPB5, has cytoprotective effects under inflammatory conditions. Advanced glycation end-products (AGE) are produced through non-enzymatic glycation under conditions of hyperglycemia and they contribute to angiogenesis and inflammation. The aim of this study was to examine the levels of serum αB-crystallin and AGE concentrations in blood samples collected from proliferative diabetic retinopathy (PDR) patients. Blood samples were collected from seven diabetic patients with PDR and eight patients without diabetes mellitus who underwent vitrectomy due to PDR and idiopathic macular diseases, respectively, in a single center. The levels of serum αB-crystallin and AGE were measured by ELISA and correlations were assessed statistically. The serum levels (mean ± SEM) of AGE were significantly higher in PDR patients (28.41±0.46 µg/ml) than in patients with non-diabetic macular diseases (25.76±0.60 µg/ml; P=0.015), whereas there was no significant difference in serum αB-crystallin levels. There was one patient with an extremely high level of αB-crystallin, who was treated with systemic corticosteroid due to chronic autoimmune inflammatory diseases. The current prospective study showed that serum AGE levels were significantly higher in PDR patients; however, no correlations between serum AGE and αB-crystallin levels were identified. - Downregulation of AlphaB-crystallin in Retinal Pigment Epithelial Cells Exposed to Diabetes-related Stimuli In Vivo and In Vitro.
D I Wu, Satoru Kase, Y E Liu, Atsuhiro Kanda, Miyuki Murata, Susumu Ishida
In vivo (Athens, Greece), 36, 1, 132, 139, 2022, [International Magazine]
English, Scientific journal, BACKGROUND/AIM: AlphaB-crystallin plays a pivotal role in many diseases. However, the involvement of alphaB-crystallin in retinal pigment epithelial (RPE) cells with diabetes stimuli remains unknown. The aim of this study is to examine the alterations of RPE cells and alphaB-crystallin expression in diabetic models in vivo and in vitro. MATERIALS AND METHODS: Diabetic conditions in mice were induced by streptozotocin (STZ). The thickness of the RPE/choroid complex was measured by optical coherence tomography (OCT). Periodic acid-Schiff (PAS) staining was used to investigate the choriocapillaris in histological sections of murine eyeballs and oxidative stress was evaluated using immunofluorescence with anti-4-hydroxynonenal (HNE) antibody. AlphaB-crystallin expression was examined in the RPE/choroid complex using ELISA. Real-Time PCR was performed to evaluate the alphaB-crystallin expression in cultured human RPE cells with high glucose or following advanced glycation end-products (AGE) stimulation. RESULTS: In diabetic mice, OCT-based RPE/choroidal layers were thickened 2 months after STZ stimulation, where PAS-positive dilated choriocapillaris was noted. Immunoreactivity of 4-HNE was strongly observed in the RPE layer, from which a significant number of RPE cells was lost. Meanwhile, alphaB-crystallin expression in 2-month STZ mice was significantly lower compared to controls. In accordance with these results, in vitro data showed that the alphaB-crystallin expression was also significantly lower in RPE cells with high glucose or following AGE stimulation compared to untreated cells. CONCLUSION: In both types of diabetic models the expression of alphaB-crystallin was found to be downregulated in RPE cells and was associated with increased levels of oxidative stress. - A Novel Deep Learning Architecture for Vascular Area Measurement in Fundus Images
Kanae Fukutsu, Michiyuki Saito, Kousuke Noda, Miyuki Murata, Satoru Kase, Ryosuke Shiba, Naoki Isogai, Yoshikazu Asano, Nagisa Hanawa, Mitsuru Dohke, Manabu Kase, Susumu Ishida
Ophthalmology Science, 100004, 100004, Elsevier BV, Feb. 2021
Scientific journal - ROCK1 Mediates Retinal Glial Cell Migration Promoted by Acrolein.
Kanae Fukutsu, Miyuki Murata, Kasumi Kikuchi, Shiho Yoshida, Kousuke Noda, Susumu Ishida
Frontiers in medicine, 8, 717602, 717602, 2021, [International Magazine]
English, Scientific journal, Objective: Acrolein is a highly reactive aldehyde that covalently binds to cellular macromolecules and subsequently modulates cellular function. Our previous study demonstrated that acrolein induces glial cell migration, a pathological hallmark of diabetic retinopathy; however, the detailed cellular mechanism remains unclear. The purpose of this study was to investigate the role of acrolein in retinal glial cell migration by focusing on rho-associated coiled-coil-containing protein kinases (ROCKs). Methods: Immunofluorescence staining for ROCK isoforms was performed using sections of fibrovascular tissue obtained from the eyes of patients with proliferative diabetic retinopathy (PDR). Rat retinal Müller glial cell line, TR-MUL5, was stimulated with acrolein and the levels of ROCK1 were evaluated using real-time PCR and western blotting. Phosphorylation of the myosin-binding subunit of myosin light chain phosphatase [myosin phosphatase target subunit 1, (MYPT1)] and myosin light chain 2 (MLC2) was assessed. The cell migration rate of TR-MUL5 cells exposed to acrolein and/or ripasudil, a non-selective ROCK inhibitor, was measured using the Oris cell migration assay. Results: ROCK isoforms, ROCK1 and ROCK2, were positively stained in the cytosol of glial cells in fibrovascular tissues. In TR-MUL5 cells, the mRNA expression level of Rock1, but not Rock2, was increased following acrolein stimulation. In line with the PCR data, western blotting showed increase in ROCK1 and cleaved ROCK1 protein in TR-MUL5 cells stimulated with acrolein. N-acetylcysteine (NAC) suppressed acrolein-associated Rock1 upregulation in TR-MUL5 cells. Acrolein augmented the phosphorylation of MYPT1 and MLC2 and increased the cell migration rate of TR-MUL5 cells, both of which were abrogated by ripasudil. Conclusions: Our study demonstrated that ROCK1 mediates the migration of retinal glial cells promoted by the unsaturated aldehyde acrolein. - Involvement of Müller Glial Autoinduction of TGF-β in Diabetic Fibrovascular Proliferation Via Glial-Mesenchymal Transition.
Di Wu, Atsuhiro Kanda, Ye Liu, Kousuke Noda, Miyuki Murata, Susumu Ishida
Investigative ophthalmology & visual science, 61, 14, 29, 29, 01 Dec. 2020, [International Magazine]
English, Scientific journal, Purpose: Müller glial-mesenchymal transition (GMT) is reported as the fibrogenic mechanism promoted by TGF-β-SNAIL axis in Müller cells transdifferentiated into myofibroblasts. Here we show the multifaceted involvement of TGF-β in diabetic fibrovascular proliferation via Müller GMT and VEGF-A production. Methods: Surgically excised fibrovascular tissues from the eyes of patients with proliferative diabetic retinopathy were processed for immunofluorescence analyses of TGF-β downstream molecules. Human Müller glial cells were used to evaluate changes in gene and protein expression with real-time quantitative PCR and ELISA, respectively. Immunoblot analyses were performed to detect TGF-β signal activation. Results: Müller glial cells in patient fibrovascular tissues were immunopositive for GMT-related molecular markers, including SNAIL and smooth muscle protein 22, together with colocalization of VEGF-A and TGF-β receptors. In vitro administration of TGF-β1/2 upregulated TGFB1 and TGFB2, both of which were suppressed by inhibitors for nuclear factor-κB, glycogen synthase kinase-3, and p38 mitogen-activated protein kinase. Of the various profibrotic cytokines, TGF-β1/2 application exclusively induced Müller glial VEGFA mRNA expression, which was decreased by pretreatment with small interfering RNA for SMAD2 and inhibitors for p38 mitogen-activated protein kinase and phosphatidylinositol 3-kinase. Supporting these findings, TGF-β1/2 stimulation to Müller cells increased the phosphorylation of these intracellular signaling molecules, all of which were also activated in Müller glial cells in patient fibrovascular tissues. Conclusions: This study underscored the significance of Müller glial autoinduction of TGF-β as a pathogenic cue to facilitate diabetic fibrovascular proliferation via TGF-β-driven GMT and VEGF-A-driven angiogenesis. - 増殖糖尿病網膜症患者の硝子体中アクロレイン結合蛋白と単球走化性因子
村田 美幸, 福津 佳苗, 野田 航介, 吉田 志帆, 神田 敦宏, 石田 晋
糖尿病合併症, 34, Suppl.1, 262, 262, (一社)日本糖尿病合併症学会, Nov. 2020
Japanese - 機械学習によって自動算出された網膜動静脈面積と血圧脈波の関係
福津 佳苗, 齋藤 理幸, 野田 航介, 村田 美幸, 加瀬 諭, 柴 涼介, 磯貝 直己, 道家 充, 石田 晋
眼科臨床紀要, 13, 10, 683, 684, 眼科臨床紀要会, Oct. 2020
Japanese - Blockade of Platelet-Derived Growth Factor Signaling Inhibits Choroidal Neovascularization and Subretinal Fibrosis in Mice.
Ye Liu, Kousuke Noda, Miyuki Murata, Di Wu, Atsuhiro Kanda, Susumu Ishida
Journal of clinical medicine, 9, 7, 15 Jul. 2020, [International Magazine]
English, Scientific journal, Neovascular age related macular degeneration (nAMD) leads to severe vision loss worldwide and is characterized by the formation of choroidal neovascularization (CNV) and fibrosis. In the current study, we aimed to investigate the effect of blockade for platelet derived growth factor receptor-β (PDGFR-β) on the formation of choroidal neovascularization and fibrosis in the laser-induced CNV model in mice. Firstly, the presence of PDGFR-β in CNV lesions were confirmed. Intravitreal injection of PDGFR-β neutralizing antibody significantly reduced the size of CNV and subretinal fibrosis. Additionally, subretinal hyperreflective material (SHRM), a landmark feature on OCT as a risk factor for subretinal fibrosis formation in nAMD patients was also suppressed by PDGFR-β blockade. Furthermore, pericytes were abundantly recruited to the CNV lesions during CNV formation, however, blockade of PDGFR-β significantly reduced pericyte recruitment. In addition, PDGF-BB stimulation increased the migration of the rat retinal pericyte cell line, R-rPCT1, which was abrogated by the neutralization of PDGFR-β. These results indicate that blockade of PDGFR-β attenuates laser-induced CNV and fibrosis through the inhibition of pericyte migration. - Regulation of Spermine Oxidase through Hypoxia-Inducible Factor-1α Signaling in Retinal Glial Cells under Hypoxic Conditions.
Di Wu, Kousuke Noda, Miyuki Murata, Ye Liu, Atsuhiro Kanda, Susumu Ishida
Investigative ophthalmology & visual science, 61, 6, 52, 52, 03 Jun. 2020, [International Magazine]
English, Scientific journal, Purpose: Acrolein, a highly reactive unsaturated aldehyde, is known to facilitate glial cell migration, one of the pathological hallmarks in diabetic retinopathy. However, cellular mechanisms of acrolein generation in retinal glial cells remains elusive. In the present study, we investigated the role and regulation of spermine oxidase (SMOX), one of the enzymes related to acrolein generation, in retinal glial cells under hypoxic condition. Methods: Immunofluorescence staining for SMOX was performed using sections of fibrovascular tissues obtained from patients with proliferative diabetic retinopathy. Expression levels of polyamine oxidation enzymes including SMOX were analyzed in rat retinal Müller cell line 5 (TR-MUL5) cells under either normoxic or hypoxic conditions. The transcriptional activity of Smox in TR-MUL5 cells was evaluated using the luciferase assay. Levels of acrolein-conjugated protein, Nε-(3-formyl-3,4-dehydropiperidino) lysine adduct (FDP-Lys), and hydrogen peroxide were measured. Results: SMOX was localized in glial cells in fibrovascular tissues. Hypoxia induced SMOX production in TR-MUL5 cells, which was suppressed by silencing of hypoxia-inducible factor-1α (Hif1a), but not Hif2a. Transcriptional activity of Smox was regulated through HIF-1 binding to hypoxia response elements 2, 3, and 4 sites in the promoter region of Smox. Generation of FDP-Lys and hydrogen peroxide increased in TR-MUL5 cells under hypoxic condition, which was abrogated by SMOX inhibitor MDL72527. Conclusions: The current data demonstrated that hypoxia regulates production of SMOX, which plays a role in the generation of oxidative stress inducers, through HIF-1α signaling in Müller glial cells under hypoxic condition. - Glucocorticoid-transactivated TSC22D3 attenuates hypoxia- and diabetes-induced Müller glial galectin-1 expression via HIF-1α destabilization.
Atsuhiro Kanda, Ikuyo Hirose, Kousuke Noda, Miyuki Murata, Susumu Ishida
Journal of cellular and molecular medicine, 24, 8, 4589, 4599, Apr. 2020, [International Magazine]
English, Scientific journal, Galectin-1/LGALS1, a newly recognized angiogenic factor, contributes to the pathogenesis of diabetic retinopathy (DR). Recently, we demonstrated that glucocorticoids suppressed an interleukin-1β-driven inflammatory pathway for galectin-1 expression in vitro and in vivo. Here, we show glucocorticoid-mediated inhibitory mechanism against hypoxia-inducible factor (HIF)-1α-involved galectin-1 expression in human Müller glial cells and the retina of diabetic mice. Hypoxia-induced increases in galectin-1/LGALS1 expression and promoter activity were attenuated by dexamethasone and triamcinolone acetonide in vitro. Glucocorticoid application to hypoxia-stimulated cells decreased HIF-1α protein, but not mRNA, together with its DNA-binding activity, while transactivating TSC22 domain family member (TSC22D)3 mRNA and protein expression. Co-immunoprecipitation revealed that glucocorticoid-transactivated TSC22D3 interacted with HIF-1α, leading to degradation of hypoxia-stabilized HIF-1α via the ubiquitin-proteasome pathway. Silencing TSC22D3 reversed glucocorticoid-mediated ubiquitination of HIF-1α and subsequent down-regulation of HIF-1α and galectin-1/LGALS1 levels. Glucocorticoid treatment to mice significantly alleviated diabetes-induced retinal HIF-1α and galectin-1/Lgals1 levels, while increasing TSC22D3 expression. Fibrovascular tissues from patients with proliferative DR demonstrated co-localization of galectin-1 and HIF-1α in glial cells partially positive for TSC22D3. These results indicate that glucocorticoid-transactivated TSC22D3 attenuates hypoxia- and diabetes-induced retinal glial galectin-1/LGALS1 expression via HIF-1α destabilization, highlighting therapeutic implications for DR in the era of anti-vascular endothelial growth factor treatment. - SDT fattyラット水晶体におけるポリオール経路と酸化ストレスの継時的解析
菊地 香澄, 村田 美幸, 野田 航介, 神田 敦宏, 加瀬 諭, 影山 靖, 篠原 雅巳, 笹瀬 智彦, 石田 晋
日本眼科学会雑誌, 124, 臨増, 242, 242, (公財)日本眼科学会, Mar. 2020
Japanese - アクロレインによる網膜ミュラーグリア細胞の遊走作用とROCK1の関与
福津 佳苗, 村田 美幸, 野田 航介, 吉田 志帆, 神田 敦宏, 石田 晋
日本眼科学会雑誌, 124, 臨増, 275, 275, (公財)日本眼科学会, Mar. 2020
Japanese - Pathological Role of Unsaturated Aldehyde Acrolein in Diabetic Retinopathy.
Miyuki Murata, Kousuke Noda, Susumu Ishida
Frontiers in immunology, 11, 589531, 589531, 2020, [International Magazine]
English, Scientific journal, With increasing prevalence of diabetes and a progressively aging society, diabetic retinopathy is emerging as one of the global leading causes of blindness. Recent studies have shown that vascular endothelial growth factor (VEGF) plays a central role in the pathogenesis of diabetic retinopathy and anti-VEGF agents have become the first-line therapy for the vision-threatening disease. However, recent studies have also demonstrated that diabetic retinopathy is a multifactorial disease and that VEGF-independent mechanism(s) also underlie much of the pathological changes in diabetic retinopathy. Acrolein is a highly reactive unsaturated aldehyde and is implicated in protein dysfunction. As acrolein is common in air pollutants, previous studies have focused on it as an exogenous causative factor, for instance, in the development of respiratory diseases. However, it has been discovered that acrolein is also endogenously produced and induces cell toxicity and oxidative stress in the body. In addition, accumulating evidence suggests that acrolein and/or acrolein-conjugated proteins are associated with the molecular mechanisms in diabetic retinopathy. This review summarizes the pathological roles and mechanisms of endogenous acrolein production in the pathogenesis of diabetic retinopathy. - Cytoprotective Effect of Astaxanthin in a Model of Normal Intraocular Pressure Glaucoma.
Kasumi Kikuchi, Zhenyu Dong, Yasuhiro Shinmei, Miyuki Murata, Atsuhiro Kanda, Kosuke Noda, Takayuki Harada, Susumu Ishida
Journal of ophthalmology, 2020, 9539681, 9539681, 2020, [International Magazine]
English, Scientific journal, Glaucoma is characterized by axonal degeneration of retinal ganglion cells (RGCs) and apoptotic death of their cell bodies. Lowering intraocular pressure is currently the only way to treat glaucoma, but it is often insufficient to inhibit the progression of the disease. Glaucoma is a multifactorial disease, and the involvement of oxidative stress has recently received much attention. In the present study, we investigated the cytoprotective effect of astaxanthin (AST) on RGC degeneration using a normal-tension glaucoma (NTG) mouse model, which lacks the glutamate/aspartate transporter (Glast) and demonstrates spontaneous RGC and optic nerve degeneration without elevated intraocular pressure. Three-week-old Glast± mice were given intraperitoneal injections of AST at 10, 30, or 60 mg/kg/day or vehicle alone, and littermate control mice were given vehicle alone for 14 days, respectively. Five weeks after birth, the number of RGCs was counted in paraffin sections of retinal tissues stained with hematoxylin and eosin. We also used a retrograde labeling technique to quantify the number of RGCs. Additionally, the phosphorylated (p) IκB/total IκB ratio and the 4-hydroxynonenal (HNE) were measured in retinal tissues. The number of RGCs in Glast± mice was significantly decreased compared with that in control mice. RGC loss was suppressed by the administration of AST at 60 mg/kg/day, compared with vehicle alone. Following AST administration, the concentration of 4-HNE in the retina was also suppressed, but the pIκB/IκB ratio did not change. Our study revealed that the antioxidative stress effects of AST inhibit RGC degeneration in the retina and may be useful in the treatment of NTG. - Diabetic Cataract in Spontaneously Diabetic Torii Fatty Rats.
Kasumi Kikuchi, Miyuki Murata, Kousuke Noda, Satoru Kase, Yoshiaki Tagawa, Yasushi Kageyama, Masami Shinohara, Tomohiko Sasase, Susumu Ishida
Journal of diabetes research, 2020, 3058547, 3058547, 2020, [International Magazine]
English, Scientific journal, Spontaneously Diabetic Torii (SDT) fatty rat is a novel animal model of type 2 diabetes with obesity. SDT fatty rats develop hyperglycemia, dyslipidemia, and other diabetic complications including ocular disorders; however, diabetic cataract formation in SDT fatty rats has not been fully investigated. The aim of the current study was to investigate the characteristics of cataract in the SDT fatty rats. The mean body weight of SDT fatty rats is larger than that of age-matched Sprague-Dawley (SD) rats and control animals until 8 weeks of age, and thereafter the growing speed decreased until the end of observation at 16 weeks of age. Blood glucose levels in SDT fatty rats were significantly higher than those in SD rats throughout the observational period. Slit-lamp examination revealed that no rats showed cataract formation at 5 weeks of age; however, SDT fatty rats gradually developed cortical cataract and posterior subcapsular cataract, both of which are the common types of cataract in patients with type 2 diabetes. The levels of glucose, sorbitol, and fructose were higher in the lens tissues of SDT fatty rats in comparison with that of SD rats. Furthermore, the level of 4-hydroxynonenal (4-HNE) was higher in the lens of SDT fatty rats than in that of SD rats. By contrast, total glutathione (GSH) concentration was lower in the lens of SDT fatty rats than in that of SD rats. The present study demonstrated that the cataractogenesis in SDT fatty rats resembled human diabetic cataract formation, indicating that SDT fatty rats serve as a potential animal model in researches on human cataract associated with type 2 diabetes and obesity. - 網膜色素上皮細胞における胎盤成長因子のアポトーシス抑制効果
村田 美幸, 野田 航介, 長谷 敬太郎, 神田 敦宏, 石田 晋
眼科臨床紀要, 12, 11, 850, 850, 眼科臨床紀要会, Nov. 2019
Japanese - Unsaturated Aldehyde Acrolein Promotes Retinal Glial Cell Migration.
Miyuki Murata, Kousuke Noda, Shiho Yoshida, Michiyuki Saito, Akio Fujiya, Atsuhiro Kanda, Susumu Ishida
Investigative ophthalmology & visual science, 60, 13, 4425, 4435, 01 Oct. 2019, [International Magazine]
English, Scientific journal, Purpose: To investigate the effect of the unsaturated aldehyde acrolein on retinal glial cell migration. Methods: Müller glial cell markers expression in TR-MUL5 were confirmed by RT-PCR and immunostaining. Cell viability and migration rate of TR-MUL5 cells were assessed after the stimulation with acrolein. DNA microarray analysis was performed to analyze changes in the expression levels of migration-related genes in Müller glial cells stimulated with acrolein. Real-time PCR and ELISA were performed to validate DNA microarray analysis results. Inhibitors of C-X-C motif chemokine ligand 1 (CXCL1), one of the genes highly upregulated after the exposure to acrolein, and blockers of its receptor, CXCR2, were used to investigate the role of the CXCL1-CXCR2 axis on glial cell migration. CXCL1 concentration was measured in vitreous fluid samples obtained from proliferative diabetic retinopathy (PDR) and nondiabetic control eyes. CXCL1 and CXCR2 expression in glial cells of fibrovascular tissues obtained from PDR patients was examined by immunostaining. Results: At a high concentration, acrolein (100 μM) significantly decreased cell viability. However, in moderate, sublethal concentrations (25-50 μM), acrolein induced cell migration and substantially increased the production of CXCL1 in TR-MUL5 cells. CXCL1 concentration was significantly elevated in vitreous fluids of PDR patients, and CXCL1 and CXCR2 were present in glial cells in fibrovascular tissues of PDR patients. CXCL1 stimulation increased glial cell migration in a dose-dependent manner, which was abrogated by the neutralization of the CXCL1-CXCR2 axis. Conclusions: Our data demonstrate that acrolein promotes retinal Müller glial cell migration by enhancing CXCL1 production. - Spontaneously Diabetic Torii fattyラット網膜における炎症性サイトカインの発現解析
菊地 香澄, 野田 航介, 村田 美幸, 神田 敦宏, 加瀬 諭, 影山 靖, 篠原 雅巳, 笹瀬 智彦, 石田 晋
糖尿病合併症, 33, Suppl.1, 271, 271, (一社)日本糖尿病合併症学会, Sep. 2019
Japanese - 網膜色素上皮細胞における胎盤成長因子による血管内皮増殖因子受容体 2の安定化機序
村田 美幸, 野田 航介, 長谷 敬太郎, Di Wu, 神田 敦宏, 石田 晋
糖尿病合併症, 33, Suppl.1, 271, 271, (一社)日本糖尿病合併症学会, Sep. 2019
Japanese - Spontaneously Diabetic Torii fattyラットにおける糖尿病白内障の検討
菊地 香澄, 野田 航介, 村田 美幸, 田川 義晃, 神田 敦宏, 加瀬 諭, 影山 靖, 篠原 雅巳, 笹瀬 智彦, 石田 晋
日本眼科学会雑誌, 123, 臨増, 166, 166, (公財)日本眼科学会, Mar. 2019
Japanese - A novel pathway of LPS uptake through syndecan-1 leading to pyroptotic cell death.
Shigetoshi Yokoyama, Yan Cai, Miyuki Murata, Takeshi Tomita, Mitsuhiro Yoneda, Lei Xu, Aprile L Pilon, Raul E Cachau, Shioko Kimura
eLife, 7, 07 Dec. 2018, [International Magazine]
English, Scientific journal, Intracellular lipopolysaccharide (LPS) triggers the non-canonical inflammasome pathway, resulting in pyroptosis of innate immune cells. In addition to its well-known proinflammatory role, LPS can directly cause regression of some tumors, although the underlying mechanism has remained unknown. Here we show that secretoglobin(SCGB)3A2, a small protein predominantly secreted in airways, chaperones LPS to the cytosol through the cell surface receptor syndecan-1; this leads to pyroptotic cell death driven by caspase-11. SCGB3A2 and LPS co-treatment significantly induced pyroptosis of macrophage RAW264.7 cells and decreased cancer cell proliferation in vitro, while SCGB3A2 treatment resulted in reduced progression of xenograft tumors in mice. These data suggest a conserved function for SCGB3A2 in the innate immune system and cancer cells. These findings demonstrate a critical role for SCGB3A2 as an LPS delivery vehicle; they reveal one mechanism whereby LPS enters innate immune cells leading to pyroptosis, and they clarify the direct effect of LPS on cancer cells. - 血管径自動解析プログラムによる網膜動静脈比と血圧脈波の相関
齋藤 理幸, 野田 航介, 星川 靖裕, 小林 正彦, 道家 充, 村田 美幸, 齋藤 航, 加瀬 学, 石田 晋
眼科臨床紀要, 11, 11, 850, 851, 眼科臨床紀要会, Nov. 2018
Japanese - 糖尿病網膜症におけるポリアミン代謝の病態関与
野田 航介, 村田 美幸, 神田 敦宏, 石田 晋
日本糖尿病眼学会誌, 22, 132, 132, 日本糖尿病眼学会, Sep. 2018
Japanese - Proteolytic cleavage of vascular adhesion protein-1 induced by vascular endothelial growth factor in retinal capillary endothelial cells
Yoshida S, Murata M, Noda K, Matsuda T, Saito M, Saito W, Kanda A, Ishida S
Jpn J Ophthalmol, 62, 2, 256, 264, Mar. 2018, [Peer-reviewed]
English, Scientific journal - Soluble Vascular Adhesion Protein-1 Mediates Spermine Oxidation as Semicarbazide-Sensitive Amine Oxidase: Possible Role in Proliferative Diabetic Retinopathy.
Murata M, Noda K, Kawasaki A, Yoshida S, Dong Y, Saito M, Dong Z, Ando R, Mori S, Saito W, Kanda A, Ishida S
Current eye research, 42, 12, 1, 10, Sep. 2017, [Peer-reviewed], [International Magazine]
English, Scientific journal, UNLABELLED: Purpose/Aim of the study: To explore the possible role of vascular adhesion protein-1 (VAP-1) via its enzymatic function as a semicarbazide-sensitive amine oxidase (SSAO) in the pathogenesis of proliferative diabetic retinopathy (PDR). MATERIALS AND METHODS: The levels of soluble VAP-1/SSAO and the unsaturated aldehyde acrolein (ACR)-conjugated protein, Nε-(3-formyl-3, 4-dehydropiperidino) lysine adduct (FDP-Lys), were measured in vitreous fluid samples of PDR and non-diabetic patients using ELISA. Recombinant human VAP-1/SSAO (rhVAP-1/SSAO) was incubated with spermine, with or without semicarbazide or RTU-1096 (a specific inhibitor for VAP-1/SSAO). Immunofluorescence assays were performed to assess the localization of VAP-1/SSAO and FDP-Lys in fibrovascular tissues from patients with PDR. The impact of ACR on cultured retinal capillary endothelial cells was assessed using a cell viability assay and total glutathione (GSH) measurements. RESULTS: The levels of sVAP-1/SSAO and FDP-Lys were elevated in the vitreous fluid of patients with PDR. Incubation of rhVAP-1 with spermine resulted in the generation of hydrogen peroxide and FDP-Lys and the production was inhibited by semicarbazide and RTU-1096. In fibrovascular tissues, FDP-Lys and VAP-1/SSAO were present in endothelial cells. ACR stimulation reduced GSH levels in the cultured endothelial cells in a dose-dependent manner and caused cellular toxicity. CONCLUSIONS: Our results indicate the pathological role of sVAP-1/SSAO to generate hydrogen peroxide and toxic aldehyde ACR, both of which are associated with oxidative stress, as a consequence of spermine oxidation in eyes with PDR. - Vascular Adhesion Protein-1 Blockade Suppresses Ocular Inflammation After Retinal Laser Photocoagulation in Mice
Takashi Matsuda, Kousuke Noda, Miyuki Murata, Akiko Kawasaki, Atsuhiro Kanda, Yukihiko Mashima, Susumu Ishida
INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE, 58, 7, 3254, 3261, Jun. 2017, [Peer-reviewed]
English, Scientific journal - Evaluation of the Safety and Tolerability of Conjunctival Ring for Posterior Segment of the Eye
Satoshi Kinoshita, Takeshi Ohguchi, Kousuke Noda, Miyuki Murata, Shin-ichi Yasueda, Haruka Obata, Toru Matsunaga, Tsutomu Fukushima, Atsuhiro Kanda, Susumu Ishida
CURRENT EYE RESEARCH, 42, 8, 1149, 1154, 2017, [Peer-reviewed]
English, Scientific journal - Localization of Acrolein-Lysine Adduct in Fibrovascular Tissues of Proliferative Diabetic Retinopathy
Yoko Dong, Kousuke Noda, Miyuki Murata, Shiho Yoshida, Wataru Saito, Atsuhiro Kanda, Susumu Ishida
CURRENT EYE RESEARCH, 42, 1, 111, 117, Jan. 2017, [Peer-reviewed]
English, Scientific journal - Regulation of vascular endothelial growth factor-C by tumor necrosis factor- in the conjunctiva and pterygium
Yoko Dong, Satoru Kase, Zhenyu Dong, Junichi Fukuhara, Yoshiaki Tagawa, Erdal Tan Ishizuka, Miyuki Murata, Yasuhiro Shinmei, Takeshi Ohguchi, Atsuhiro Kanda, Kousuke Noda, Susumu Ishida
INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE, 38, 2, 545, 550, Aug. 2016, [Peer-reviewed]
English, Scientific journal - Phosphorylation of alphaB-crystallin in epiretinal membrane of human proliferative diabetic retinopathy.
Dong Y, Dong Z, Kase S, Ando R, Fukuhara J, Kinoshita S, Inafuku S, Tagawa Y, Ishizuka ET, Saito W, Murata M, Kanda A, Noda K, Ishida S
International journal of ophthalmology, 9, 8, 1100, 1105, 2016, [Peer-reviewed] - Alteration of N-Glycan Profiles in Diabetic Retinopathy
Saori Inafuku, Kousuke Noda, Maho Amano, Tetsu Ohashi, Chikako Yoshizawa, Wataru Saito, Miyuki Murata, Atsuhiro Kanda, Shin-Ichiro Nishimura, Susumu Ishida
INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE, 56, 9, 5316, 5322, Aug. 2015, [Peer-reviewed]
English, Scientific journal - Specific inhibition of serine/arginine-rich protein kinase attenuates choroidal neovascularization
Zhenyu Dong, Kousuke Noda, Atsuhiro Kanda, Junichi Fukuhara, Ryo Ando, Miyuki Murata, Wataru Saito, Masatoshi Hagiwara, Susumu Ishida
MOLECULAR VISION, 19, 536, 543, Mar. 2013, [Peer-reviewed]
English, Scientific journal - Tissue Kallikrein Attenuates Choroidal Neovascularization via Cleavage of Vascular Endothelial Growth Factor
Junichi Fukuhara, Kousuke Noda, Miyuki Murata, Shiho Namba, Satoshi Kinoshita, Zhenyu Dong, Ryo Ando, Anton Lennikov, Atsuhiro Kanda, Susumu Ishida
INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE, 54, 1, 274, 279, Jan. 2013, [Peer-reviewed]
English, Scientific journal - ALPHAB-CRYSTALLIN EXPRESSION IN EPIRETINAL MEMBRANE OF HUMAN PROLIFERATIVE DIABETIC RETINOPATHY
Zhenyu Dong, Satoru Kase, Ryo Ando, Junichi Fukuhara, Wataru Saito, Atsuhiro Kanda, Miyuki Murata, Kousuke Noda, Susumu Ishida
RETINA-THE JOURNAL OF RETINAL AND VITREOUS DISEASES, 32, 6, 1190, 1196, Jun. 2012, [Peer-reviewed]
English, Scientific journal - Soluble Vascular Adhesion Protein-1 Accumulates in Proliferative Diabetic Retinopathy
Miyuki Murata, Kousuke Noda, Junichi Fukuhara, Atsuhiro Kanda, Satoru Kase, Wataru Saito, Yoko Ozawa, Satsuki Mochizuki, Shioko Kimura, Yukihiko Mashima, Yasunori Okada, Susumu Ishida
INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE, 53, 7, 4055, 4062, Jun. 2012, [Peer-reviewed]
English, Scientific journal - Amelioration of ultraviolet-induced photokeratitis in mice treated with astaxanthin eye drops
Anton Lennikov, Nobuyoshi Kitaichi, Risa Fukase, Miyuki Murata, Kousuke Noda, Ryo Ando, Takeshi Ohguchi, Tetsuya Kawakita, Shigeaki Ohno, Susumu Ishida
MOLECULAR VISION, 18, 49, 455, 464, Feb. 2012, [Peer-reviewed]
English, Scientific journal - Immunolocalization of Vascular Adhesion Protein-1 in Human Conjunctival Tumors
Junichi Fukuhara, Satoru Kase, Kousuke Noda, Miyuki Murata, Mika Noda, Ryo Ando, Zhenyu Dong, Atsuhiro Kanda, Susumu Ishida
OPHTHALMIC RESEARCH, 48, 1, 33, 37, 2012, [Peer-reviewed]
English, Scientific journal - Tissue factor expression in human pterygium
Ryo Ando, Satoru Kase, Tsutomu Ohashi, Zhenyu Dong, Junichi Fukuhara, Atsuhiro Kanda, Miyuki Murata, Kousuke Noda, Nobuyoshi Kitaichi, Susumu Ishida
MOLECULAR VISION, 17, 8-10, 63, 69, Jan. 2011, [Peer-reviewed]
English, Scientific journal - Requirement of STAT3 activation for maximal collagenase-1 (MMP-1) induction by epidermal growth factor and malignant characteristics in T24 bladder cancer cells
M Itoh, T Murata, T Suzuki, M Shindoh, K Nakajima, K Imai, K Yoshida
ONCOGENE, 25, 8, 1195, 1204, Feb. 2006, [Peer-reviewed]
English, Scientific journal - E1AF degradation by a ubiquitin-proteasome pathway
A Takahashi, F Higashino, M Aoyagi, K Yoshida, M Itoh, M Kobayashi, Y Totsuka, T Kohgo, M Shindoh
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 327, 2, 575, 580, Feb. 2005, [Peer-reviewed]
English, Scientific journal - EWS/ETS fusions activate telomerase in Ewing's tumors
A Takahashi, F Higashino, M Aoyagi, K Yoshida, M Itoh, S Kyo, T Ohno, T Taira, H Ariga, K Nakajima, M Hatta, M Kobayashi, H Sano, T Kohgo, M Shindoh
CANCER RESEARCH, 63, 23, 8338, 8344, Dec. 2003, [Peer-reviewed]
English, Scientific journal - Nuclear export of glucocorticoid receptor is enhanced by c-Jun N-terminal kinase-mediated phosphorylation
M Itoh, M Adachi, H Yasui, M Takekawa, H Tanaka, K Imai
MOLECULAR ENDOCRINOLOGY, 16, 10, 2382, 2392, Oct. 2002, [Peer-reviewed]
English, Scientific journal - Involvement of p27(KIP1) degradation by Skp2 in the regulation of proliferation in response to wounding of corneal epithelium
K Yoshida, K Nakayama, H Nagahama, T Harada, C Harada, J Imaki, A Matsuda, K Yamamoto, M Ito, S Ohno, K Nakayama
INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE, 43, 2, 364, 370, Feb. 2002, [Peer-reviewed]
English, Scientific journal
Other Activities and Achievements
- ニューラルネットワークによる眼底写真の網膜動静脈検出
福津佳苗, 齋藤理幸, 野田航介, 村田美幸, 加瀬諭, 柴涼介, 磯貝直己, 道家充, 石田晋, 日本眼循環学会講演抄録集, 36th, 2019 - 眼科のトランスレーショナルリサーチ 多機能蛋白質に着目した糖尿病網膜症に対する創薬研究
野田 航介, 村田 美幸, 稲福 沙織, 松田 剛, 吉田 志帆, 董 陽子, 木下 哲志, 安藤 亮, 藤谷 顕雄, 齋藤 理幸, 董 震宇, 森 祥平, 加瀬 諭, 吉澤 史子, 齋藤 航, 神田 敦宏, 石田 晋, 眞島 行彦, 笹瀬 智彦, 天野 麻穂, 大橋 哲, 西村 伸一郎, 今川 貴仁, 白仁田 明生, 日本眼科学会雑誌, 122, 3, 223, 248, Mar. 2018
(公財)日本眼科学会, Japanese - 増殖糖尿病網膜症患者の硝子体中アクロレイン結合蛋白FDP-lysine濃度
森 祥平, 村田 美幸, 野田 航介, 鈴木 智浩, 齋藤 理幸, 安藤 亮, 加瀬 諭, 齋藤 航, 神田 敦宏, 石田 晋, 眼科臨床紀要, 10, 11, 920, 921, Nov. 2017
眼科臨床紀要会, Japanese - 網膜グリア細胞に対するアクロレインの作用
野田 航介, 村田 美幸, 吉田 志帆, 神田 敦宏, 石田 晋, 糖尿病合併症, 31, Suppl.1, 347, 347, Oct. 2017
(一社)日本糖尿病合併症学会, Japanese - 糖尿病網膜症におけるポリアミン代謝の病態関与
野田 航介, 村田 美幸, 神田 敦宏, 石田 晋, 糖尿病合併症, 30, Suppl.1, 248, 248, Sep. 2016
(一社)日本糖尿病合併症学会, Japanese - 糖尿病網膜症の硝子体および血漿におけるN型糖鎖プロファイル
稲福 沙織, 野田 航介, 天野 麻穂, 大橋 哲, 齋藤 航, 村田 美幸, 神田 敦宏, 西村 紳一郎, 石田 晋, 日本糖尿病眼学会誌, 20, 114, 114, Mar. 2016
日本糖尿病眼学会, Japanese - Alteration of N-glycan Profiles in Diabetic Retinopathy
Saori Inafuku, Kousuke Noda, Maho Amano, Miyuki Murata, Wataru Saito, Tetsu Ohashi, Atsuhiro Kanda, Shin-Ichiro Nishimura, Susumu Ishida, INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE, 56, 7, Jun. 2015
English, Summary international conference - 糖尿病網膜症の硝子体および血漿におけるN型糖鎖の網羅的解析
稲福 沙織, 野田 航介, 天野 麻穂, 大橋 哲, 吉澤 史子, 齋藤 航, 村田 美幸, 神田 敦宏, 西村 紳一郎, 石田 晋, 日本眼科学会雑誌, 119, 臨増, 160, 160, Mar. 2015
(公財)日本眼科学会, Japanese - 糖負荷刺激によるシアル酸転移酵素の誘導
稲福 沙織, 野田 航介, 天野 麻穂, 大橋 哲, 齋藤 航, 村田 美幸, 神田 敦宏, 西村 紳一郎, 石田 晋, 日本眼科学会雑誌, 119, 臨増, 199, 199, Mar. 2015
(公財)日本眼科学会, Japanese - SRPK阻害薬による脈絡膜血管新生の抑制効果
董 震宇, 野田 航介, 斉藤 航, 木下 哲志, 福原 淳一, 安藤 亮, 村田 美幸, 神田 敦宏, 萩原 正敏, 石田 晋, 日本眼科学会雑誌, 117, 臨増, 289, 289, Mar. 2013
(公財)日本眼科学会, Japanese - 増殖糖尿病網膜症におけるVEGF阻害剤の硝子体内VAP-1濃度への影響
安藤 亮, 野田 航介, 村田 美幸, 難波 志帆, 木下 哲志, 福原 淳一, 董 震宇, アントン・レニコフ, 齋藤 航, 神田 敦宏, 石田 晋, 日本眼科学会雑誌, 116, 臨増, 265, 265, Mar. 2012
(公財)日本眼科学会, Japanese - ヒト翼状片におけるTissue Factorの発現
安藤 亮, 加瀬 諭, 大橋 勉, 董 震宇, 福原 淳一, アントン・レニコフ, 神田 敦宏, 村田 美幸, 野田 航介, 石田 晋, 日本眼科学会雑誌, 115, 臨増, 203, 203, Apr. 2011
(公財)日本眼科学会, Japanese - アスタキサンチン点眼による紫外線急性角膜炎の軽症化
アントン・レニコフ, 北市 伸義, 深瀬 理沙, 村田 美幸, 野田 航介, 大野 重昭, 石田 晋, 日本眼科学会雑誌, 115, 臨増, 227, 227, Apr. 2011
(公財)日本眼科学会, Japanese - 増殖糖尿病網膜症の線維血管膜におけるalphaB-crystallinの発現
董 震宇, 加瀬 諭, 安藤 亮, 福原 淳一, アントン・レニコフ, 神田 敦宏, 村田 美幸, 野田 航介, 斎藤 航, 石田 晋, 日本眼科学会雑誌, 115, 臨増, 231, 231, Apr. 2011
(公財)日本眼科学会, Japanese - 増殖糖尿病網膜症患者の硝子体液におけるVAP-1濃度
福原 淳一, 野田 航介, 村田 美幸, 齋藤 航, 董 震宇, 安藤 亮, アントン・レニコフ, 神田 敦宏, 石田 晋, 日本眼科学会雑誌, 115, 臨増, 231, 231, Apr. 2011
(公財)日本眼科学会, Japanese
Affiliated academic society
Research Themes
- 滲出型加齢黄斑変性の網膜色素上皮障害における毒性アルデヒドアクロレインの関与
科学研究費助成事業
01 Apr. 2024 - 31 Mar. 2027
村田 美幸, 野田 航介
日本学術振興会, 基盤研究(C), 北海道大学, 24K12777 - Role of alphaB-crystallin in epithelial-mesenchymal transition in retinal Muller cells
Grants-in-Aid for Scientific Research
01 Apr. 2024 - 31 Mar. 2027
加瀬 諭, 村田 美幸
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 24K12797 - 加齢黄斑変性におけるセリン/スレオニンキナーゼLRRK2の病態意義解明
科学研究費助成事業
01 Apr. 2021 - 31 Mar. 2025
野田 航介, 村田 美幸, 矢部 一郎, 加瀬 諭
加齢黄斑変性(AMD)の病態基盤には網膜色素上皮細胞(RPE)の変性や細胞死が関与している。興味深いことに、AMD患者ではパーキンソン病の発症リスクが有意に高いとする報告が近年相次いでおり、パーキンソン病が神経変性疾患であることから、この2つの疾患には共通病態が存在する可能性があると仮説を立てた。本研究の目的は、「AMDにおけるパーキンソン病との共通病態を探索すること」である。
研究開始初年度は、まずヒト摘出眼球切片を用いてRPEにおけるαシヌクレインおよびリン酸化αシヌクレインの局在を免疫組織染色によって検討した。その結果、ヒト眼球組織のRPEにおいてαシヌクレインとリン酸化αシヌクレインの比較的強い染色シグナルが確認された。次に、RPEにおけるαシヌクレインおよびリン酸化αシヌクレインの産生機序を検討するため、in vitro実験系を用いて検討を行なった。血清除去条件下でヒト培養RPE株hTERT RPE-1を24時間培養した後のαシヌクレインおよびリン酸化αシヌクレインの変化をwestern blottingで検討した。その結果、血清除去によってhTERT RPE-1におけるαシヌクレインおよびリン酸化αシヌクレインのタンパク量は増加していた。また同条件下では、パーキンソン病発症に重要な役割を演じているとされるleucine-rich repeat kinase 2 (LRRK2)のmRNA発現およびタンパク量も増加した。
リン酸化αシヌクレインはオリゴマーを形成することで神経細胞の変性を引き起こすとされる。また、αシヌクレインのリン酸化およびその蓄積には前述のLRRK2が関与するとする報告がある。以上のことは、パーキンソン病と同様に、加齢などの要因によるRPEの変性にもリン酸化αシヌクレインが関与している可能性を示している。
日本学術振興会, 基盤研究(B), 北海道大学, 21H03091 - 糖尿病網膜症病態におけるアクロレインの炎症誘導機構
科学研究費助成事業
01 Apr. 2021 - 31 Mar. 2024
村田 美幸, 野田 航介
糖尿病網膜症は重篤な視機能障害をきたすため、その病態の解明は眼科学における重要な課題である。我々は過去に、増殖糖尿病網膜症患者の 線維血管組織において網膜グリア細胞に不飽和アルデヒドの一種であるアクロレインが結合した蛋白が蓄積していることを報告した。アクロレ インは反応性が高く、様々な蛋白と結合することでその機能異常を惹起するため、神経変性疾患や悪性腫瘍など多様な疾患病態で注目されてい る分子である。本研究では、糖尿病網膜症におけるアクロレインの炎症誘導機構について検討をおこなう。
昨年度までに、アクロレインがラット培養網膜グリア細胞株TR-MUL5において、単球走化性因子(monocyte chemoattractant protein-1、MCP-1)を含む複数の炎症関連分子の発現を誘導することが明らかになった。アクロレインによるMCP-1の発現誘導は酸化ストレス阻害剤であるN-acetyl-cysteine の添加により抑制されたことから、アクロレインによる酸化ストレスの亢進がMCP-1の誘導に関わることが示唆された。トランスウェルの下方にTR-MUL5を播種し、アクロレインを添加して24時間後に上層のウェルにマクロファージ細胞株RAW264.7を播種したところ、アクロレイン無添加群に比べ有意にRAW264.7の遊走の亢進が認められた。MCP-1の受容体であるCCR2の阻害剤をRAW264.7に添加したところ遊走能の亢進が抑制されたことから、アクロレイン刺激によりTR-MUL5からの分泌が増加したMCP-1が、RAW264.7の遊走を促進していることが示された。これらのことから、アクロレインに網膜グリア細胞からのMCP-1の分泌を亢進させ、同細胞へのマクロファージの遊走を促すという作用があることが明らかになった。
日本学術振興会, 基盤研究(C), 北海道大学, 21K09667 - Glycer-AGEによる網膜神経病態の機序解明と糖尿病網膜症に対する創薬
科学研究費助成事業
01 Apr. 2020 - 31 Mar. 2024
石田 晋, 村田 美幸, 神田 敦宏
糖尿病網膜症は重篤な視機能障害をきたす疾患であり、その病態理解は眼科学における重要課題である。これまでの糖尿病網膜症治療はその血管病態を制御することを目的とされてきたが、近年、糖尿病網膜症における「神経病態」が注目されている。しかし、糖尿病網膜症における神経保護的治療法は現在のところ確立されていない。
グリセルアルデヒド由来の終末糖化産物(Glycer-AGE)は、その細胞障害性の強さからToxic AGE(TAGE)とも呼ばれ、糖尿病網膜症を含む様々な疾患での病態形成への関与が報告されている。申請者グループの検討により、このTAGEが糖尿病モデルマウスの視細胞変性に寄与していることが明らかになった。本研究では、TAGEによる網膜神経病態の機序を解明し、さらにTAGEを標的とした創薬開発を行い、糖尿病網膜症の新規治療法開発を目指す。
本研究におけるこれまでの検討により、ヒト培養網膜色素上皮細胞にグリセルアルデヒドを添加してTAGE化反応を誘導し、抗TAGEモノクローナル抗体で免疫沈降をおこなって質量分析に供し、グリセルアルデヒドを添加しないコントロール群と比較することで、網膜色素上皮細胞内でTAGE化される複数のタンパク質を同定している。また、ヒト培養網膜色素上皮細胞にグリセルアルデヒドと牛血清アルブミン(BSA)を反応させて作製したTAGE化BSAを添加し、TAGE負荷が網膜色素上皮細胞機能に及ぼす影響について現在解析を進めている。
日本学術振興会, 基盤研究(B), 北海道大学, 20H03837 - The Involvement of Inflammatory Cell Migration and Unsaturated Aldehydes in the Pathogenesis of Diabetic Retinopathy
Bayer Academic Support
Apr. 2021 - Mar. 2022
Kanae Fukutsu, Miyuki Murata, Kousuke Noda
Bayer, Basic research, Department of Ophthalmology Faculty of Medicine and Graduate School of Medicine Hokkaido University, BASJ20210401022 - Mechanism of acrolein generation in diabetic retinopathy
Grants-in-Aid for Scientific Research
01 Apr. 2017 - 31 Mar. 2020
Murata Miyuki
Vascular adhesion protein-1 (VAP-1) expresses in vascular endothelial cells and involved in inflammation as a leukocyte adhesion molecule. In addition to its role as an adhesion molecule, VAP-1 also has enzymatic activity as an amine oxidase. Acrolein is a highly reactive unsaturated aldehyde and is included in cigarettes and so on as an exogenous pollutant. Recently it has been shown that acrolein is also produced endogenously as a consequence of polyamine oxidation. We found that the levels of the acrolein-conjugated protein are elevated in the vitreous fluid of patients with diabetic retinopathy (DR). VAP-1 protein levels are also elevated in the vitreous fluid of DR patients and correlated with acrolein-conjugated protein levels. In this study, we showed that VAP-1 generates acrolein by oxidizing the spermine, a kind of polyamine.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 17K11442 - 網膜の恒常性維持における胎盤増殖因子(PIGF)の役割の解析
科学研究費助成事業
2015 - 2015
村田 美幸
研究目的
血管新生や血管形成に関与する増殖因子には、血管内皮増殖因子(vascular endothelial growth factor, VEGF)-A、VEGF-B、VEGF-C、VEGF-D、VEGF-EおよびPIGF-1、PIGF-2の7種類が知られ、VEGFファミリーと呼ばれている。この中で、VEGF-Aは糖尿病網膜症や加齢黄斑変性などにおいて眼内血管新生を促し病態を悪化させる重要な因子であることが明らかとなり、VEGF-Aを標的とする阻害剤がすでに良好な治療結果をもたらしている。近年開発されたVEGF阻害薬はその標的分子も多様化し、VEGF-AのみならずPIGFを含む複数のVEGFファミリーを阻害する薬剤である。PIGFは血管新生や炎症などに関与する事が報告されているが、その生理的作用は未だ明らかになっていない。本研究では、網膜の恒常性維持に重要な働きをする網膜色素上皮細胞(retinal pigment epithelium, RPE)におけるPIGFの役割を解析した。
研究方法
培養ヒトRPE細胞にPIGF shRNAを導入して遺伝子発現を抑制し、non target shRNAを導入したコントロールと比較をおこなった。
研究結果
PIGF発現を抑制したヒトRPE株では、VEGF受容体(VEGFR)2のタンパク発現が低下していること、VEGFR2下流シグナルである細胞外シグナル調節キナーゼ(extracellular signal-regulated kinase, ERK)1/2やv-akt murine thymoma viral oncogene homolog(AKT)のリン酸化が抑制されることが明らかになった。VEGFR2のmRNA発現に変化は見られなかったことから、PIGFはVEGFR2タンパクの安定性に関与している可能性が示された。
日本学術振興会, 奨励研究, 北海道大学, 15H00579 - 網膜色素上皮細胞における上皮間葉移行関連転写因子の探索
科学研究費助成事業
2010 - 2011
村田 美幸
加齢黄斑変性(AMD)の病型の一つ、滲出型AMDの病態を特徴づける脈絡膜新生血管(CNV)はその終末期病態として線維化をきたし、視力障害の原因となる。近年,組織の線維化においてEpithelial-Mesenchymal Transition(上皮間葉移行:EMT)が着目されているが、ヒトCNV組織検体を用いた我々の検討結果ではEMTを制御する転写因子の一つSnailがヒトCNV中の網膜色素上皮細胞(RPE)に高発現しており,線維化をきたしている組織ほどその陽性率が高いことが明らかとなった。この結果はCNV中に含まれるRPEがEMTを生じていること、EMTの転写因子SnailがCNVの線維化に関与する可能性を示唆していた。
本研究では、培養ヒトRPE細胞株ARPE19を用いたin vitro実験系により、EMT関連転写因子snailの制御機構を解析した。CNV組織において発現が確認されている各種サイトカイン(transforming growth factor (TGF)-b, tumor-necrosis factor (TNF)-a, vascular endothelial growth factor (VEGF), connective tissue growth factor (CTGF)など)を用いてARPE19細胞を刺激したところ、TGF-b刺激による転写因子Snailの発現増加がRT-PCRおよび免疫組織染色によって確認された。本結果は、ヒトCNV中に存在するTGF-bがEMT転写因子Snailの発現を誘導し、CNV組織においてRPEにEMT変化を惹起するということを示唆していた。
日本学術振興会, 若手研究(B), 北海道大学, 22791644
