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Introduction. Aspergillus flavus and Fusarium keratoplasticum are common causative pathogens of fungal keratitis (FK), a severe corneal disease associated with significant morbidity and vision loss. Escalating incidence of antifungal resistance to available antifungal drugs poses a major challenge to FK treatment. Cold atmospheric plasma (CAP) is a pioneering nonpharmacologic antimicrobial intervention that has demonstrated potential as a broad-spectrum antifungal treatment.Gap statement. Previous research highlights biofilm-associated resistance as a critical barrier to effective FK treatment. Although CAP has shown promise against various fungal infections, its efficacy against biofilm and conidial forms of FK pathogens remains inadequately explored.Aim. This study aims to investigate the antifungal efficacy of CAP against clinical fungal keratitis isolates of A. flavus and F. keratoplasticum in vitro.Methodology. Power parameters (22-27 kVpp, 300-400 Hz and 20-80 mA) of a dielectric barrier discharge CAP device were optimized for inactivation of A. flavus biofilms. Optimal applied voltage and total current were applied to F. keratoplasticum biofilms and conidial suspensions of A. flavus and F. keratoplasticum. The antifungal effect of CAP treatment was investigated by evaluating fungal viability through means of metabolic activity, c.f.u. enumeration (c.f.u. ml-1) and biofilm formation.Results. For both fungal species, CAP exhibited strong time-dependent inactivation, achieving greater than 80â% reduction in metabolic activity and c.f.u. ml-1 within 300 s or less, and complete inhibition after 600 s of treatment.Conclusion. Our findings indicate that CAP is a promising broad-spectrum antifungal intervention. CAP treatment effectively reduces fungal viability in both biofilm and conidial suspension cultures of A. flavus and F. keratoplasticum, suggesting its potential as an alternative treatment strategy for fungal keratitis.
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Antifúngicos , Aspergillus flavus , Biopelículas , Fusarium , Queratitis , Gases em Plasma , Esporas Fúngicas , Aspergillus flavus/efectos de los fármacos , Fusarium/efectos de los fármacos , Biopelículas/efectos de los fármacos , Gases em Plasma/farmacología , Esporas Fúngicas/efectos de los fármacos , Antifúngicos/farmacología , Queratitis/microbiología , Infecciones Fúngicas del Ojo/microbiología , Humanos , Fusariosis/microbiología , Viabilidad Microbiana/efectos de los fármacosRESUMEN
Equine fungal keratitis represents a substantial portion of keratitis cases in horses, with fungal involvement identified in approximately half of all infectious keratitis cases. Despite its prevalence, more comprehensive retrospective analyses are needed to better understand this condition. Outcomes vary, with approximately two-thirds of cases achieving complete healing with retained vision, although enucleation is often necessary. Predominant pathogens include Aspergillus and Fusarium, with yeast reported in a minority of cases. Resistance to common antifungal agents among filamentous fungi poses a significant challenge. Advances in diagnostics, including repeat culture and antifungal susceptibility testing, as well as the incorporation of PCR technology, hold promise for improving detection and guiding treatment decisions. Newer antifungals, combination therapies, and innovative modalities such as photodynamic therapy offer hope for improved outcomes. Continued research efforts are essential to further elucidate the epidemiology, pathogenesis, and optimal management strategies for this condition.
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OBJECTIVE: To determine the predictive value of corneal ultrasound biomicroscopy (UBM) findings for the outcome of equine corneal disease. ANIMALS: 46 horses with a clinical diagnosis of either infectious ulcerative keratitis or stromal abscess. METHODS: Corneal UBM (VevoMD; UHF70; VisualSonics) of horses with infectious corneal disease presenting to the North Carolina State University Equine Ophthalmology Service from 2019 to 2023 were evaluated. Size and depth of lesion, presence of Descemet membrane disruption (DMD), corneal thickness, and aqueous humor cell counts (AHCC) were assessed. Comparisons of UBM and clinical exam findings, presence of infectious organisms, and outcome (healed or enucleated) were performed. RESULTS: The UBMs from 46 horses were evaluated. Increased AHCC was significantly associated with increased size and depth of corneal lesions on UBM but not with DMD. Deep lesions and DMD were significantly associated with an enucleation outcome. Horses treated with systemic antibiotics had significantly lower AHCC on UBM, but there were no differences in AHCC with the use of other systemic or topical medications. There was no significant correlation between infectious disease results, clinical findings (aqueous flare or cells), outcome, and UBM AHCC. CLINICAL RELEVANCE: Parameters on UBM, such as depth of lesion, DMD, and AHCC, may be useful diagnostic and prognostic tools to augment the ophthalmic exam of horses with corneal disease. The UBM findings of deep corneal lesions and DMD suggest a poor prognosis and warrant aggressive surgical intervention.
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Fungal keratitis (FK) is an invasive infection of the cornea primarily associated with Aspergillus and Fusarium species. FK is treated empirically with a limited selection of topical antifungals with varying levels of success. Though clinical infections are typically characterized by a dense network of mature mycelium, traditional models used to test antifungal susceptibility of FK isolates exclusively evaluate susceptibility in fungal cultures derived from asexual spores known as conidia. The purpose of this study was to characterize differences in fungal response when topical antifungal treatment is initiated at progressive phases of fungal development. We compared the efficacy of voriconazole and luliconazole against in vitro cultures of A. flavus and F. keratoplasticum at 0, 24, and 48 h of fungal development. A porcine cadaver corneal model was used to compare antifungal efficacy of voriconazole and luliconazole in ex vivo tissue cultures of A. flavus and F. keratoplasticum at 0, 24, and 48 h of fungal development. Our results demonstrate phase-dependent susceptibility of both A. flavus and F. keratoplasticum to both azoles in vitro as well as ex vivo. We conclude that traditional antifungal susceptibility testing with conidial suspensions does not correlate with fungal susceptibility in cultures of a more advanced developmental phase. A revised method of antifungal susceptibility testing that evaluates hyphal susceptibility may better predict fungal response in the clinical setting where treatment is often delayed until days after the initial insult.
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Congenital stationary night blindness (CSNB) is an ocular disorder characterized by nyctalopia. An autosomal recessive missense mutation in glutamate metabotropic receptor 6 (GRM6 c.533C>T, p.(Thr178Met)), called CSNB2, was previously identified in one Tennessee Walking Horse and predicted to reduce binding affinity of the neurotransmitter glutamate, impacting the retinal rod ON-bipolar cell signaling pathway. Thus, the first aim was to identify the allele frequency (AF) of CSNB2 in breeds with reported cases of CSNB and breeds closely related to the Tennessee Walking Horse. The second aim was to perform ocular examinations in multiple breeds to confirm the link between genotype and CSNB phenotype. In evaluating 3518 horses from 14 breeds, the CSNB2 allele was identified in nine previously unreported breeds. The estimated AF was highest in pacing Standardbreds (0.17) and lowest in American Quarter Horses (0.0010). Complete ophthalmic examinations and electroretinograms (ERG) were performed on 19 horses from three breeds, including one CSNB2 homozygote from each breed. All three CSNB2/CSNB2 horses had an electronegative ERG waveform under scotopic light conditions consistent with CSNB. The remaining 16 horses (seven CSNB2/N and nine N/N) had normal scotopic ERG results. All horses had normal photopic ERGs. This study provides additional evidence that GRM6 c.533C>T homozygosity is likely causal to CSNB in Tennessee Walking Horses, Standardbreds, and Missouri Fox Trotting Horses. Genetic testing is recommended for breeds with the CSNB2 allele to limit the production of affected horses. This study represents the largest across-breed identification of CSNB in the horse and suggests that this disorder is likely underdiagnosed.
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Purpose: The objective of this study is to develop and characterize electrospun corneal bandage infused with Noggin protein and evaluate its therapeutic potential in the treatment of superficial nonhealing corneal ulceration. Methods: Electrospun nanofibrous scaffolds were created with different blend ratios of polycaprolactone and gelatin and coated with different concentrations of Noggin protein. Morphologic, mechanical, degradation, and surface chemistry of the developed scaffold was assessed. Biocompatibility of the developed scaffold with corneal epithelial cells was evaluated by looking at cell viability, proliferation, and immunostaining. In vitro wound healing in the presence of Noggin-coated scaffold was evaluated by measuring wound closure rate after scratch. Results: Uniform nanofibrous scaffolds coated with Noggin were constructed through optimization of electrospinning parameters and demonstrated mechanical properties better than or similar to commercially available contact lenses used in corneal wound healing. In the presence of Noggin-coated scaffold, corneal epithelial cells showed higher proliferation and wound-healing rate. Conclusions: This Noggin-coated electrospun scaffold represents a step toward, expanding treatment options for patients with indolent corneal ulcers. Translational Relevance: In this study, the feasibility of Noggin-coated electrospun scaffold as a therapeutic for indolent corneal ulcer was evaluated. This study also provides a better perspective for understanding electrospun scaffolds as a tunable platform to infuse topical therapeutics and use as a corneal bandage.
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Lesiones de la Cornea , Andamios del Tejido , Humanos , Lentes de Contacto , Córnea , Lesiones de la Cornea/terapia , Células EpitelialesRESUMEN
The aim of this study was to assess the transmission of the ultraviolet (UV) radiation (200-400 nm) through intact enucleated globes of different species (dogs, cats, pigs, rabbits, horses, and humans) using spectrophotometry. Globes of cats (n = 6), dogs (n = 18), pigs (n = 10), rabbits (n = 6), horses (n = 10), and humans (n = 4) were analyzed. A 5-10 mm circular area of sclera and choroid from the posterior aspect of the globe was removed under a surgical microscope, leaving the retina intact in all species except the horse. Glass coverslips were added in horses and rabbits due to retinal and globe fragility. The %T of wavelengths from 200 to 800 nm were measured through the ocular media (cornea, aqueous humor, lens, and vitreous humor) and retina, and compared between species. The globes of cats and dogs allowed the most amount of UV radiation transmission, while those of pigs and humans allowed the least amount of UV radiation transmission. A small amount of UV radiation transmission through the ocular media was detected in the rabbit and horse. Results from this study will support further vision research that may be used to train companion, working, and service animals.
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Cristalino , Rayos Ultravioleta , Conejos , Caballos , Gatos , Humanos , Porcinos , Animales , Perros , Retina , Cuerpo Vítreo , CórneaRESUMEN
Ocular graft versus host disease (OGvHD) develops after allogeneic hematopoietic stem cell transplantation (HSCT) and manifests as ocular surface inflammatory disease. This study evaluated the efficacy of adeno-associated virus (AAV) gene therapy encoding human leukocyte antigen G (HLA-G) to inhibit OGvHD. A major histocompatibility mismatch chronic OGvHD murine model was evaluated. 7 days after HSCT, mice were dosed subconjunctivally with scAAV8-HLA-G1/5 (1 x 109 vg/eye), topical cyclosporine (twice daily), or left untreated. Body weights and tear production (red thread test) were recorded, and eyelid, corneal opacity, and corneal fluorescein retention were scored through day 44 after HSCT. Tissues were collected for vector biodistribution, ocular histology, and immunofluorescence. Compared with untreated HSCT eyes, those dosed with scAAV8-HLA-G1/5 had significantly reduced clinical inflammatory signs of OGvHD. On histology, eyes that received scAAV8-HLA-G1/5 or cyclosporine had a significantly lower mean limbal mononuclear cell count when compared with non-treated HSCT eyes. HLA-G immunofluorescence was detected in the subconjunctiva and peripheral cornea in HSCT animals treated with scAAV8-HLA-G1/5. Vector genomes were detected in the lacrimal gland, but not in the other tested organs. These results provide evidence that subconjunctival AAV targets ocular surface and corneal disease and support that HLA-G-based gene therapy may be an effective treatment for OGvHD.
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OBJECTIVE: The objective of the study was to determine the effect of gentamicin on CD3+ T-lymphocyte proliferation and cell viability using an in vitro cell culture model as a means of investigating the mechanism of action of low-dose intravitreal gentamicin injection. ANIMALS STUDIED: Three adult horses with no evidence of ophthalmic or systemic disease. PROCEDURE: Peripheral blood lymphocytes were treated with gentamicin at concentrations 37.5 µg/mL, 112.5 µg/mL, 187 µg/mL, 375 µg/mL, or 750 µg/mL then stimulated to proliferate with concanavalin A (ConA). 4',6-diamidino-2-phenylindole (DAPI) and carboxyfluoroscein succinimidyl ester (CSFE) were used as markers of cell viability and cell proliferation, respectively. Following 5-day culture, live cell counts and CSFE fluorescent intensity data were collected via automated cell count and flow cytometry. The experimental design was duplicated using preservative-free gentamicin and a proprietary brand formulation. Statistical analysis was performed using two-way ANOVA with Tukey's multiple comparison test. RESULTS: No statistically significant comparisons in CD3+ T-lymphocyte live cell counts and geometric mean fluorescent intensity of CSFE were identified between gentamicin concentrations or formulations. CONCLUSIONS: Gentamicin had no effect on equine peripheral blood CD3+ T-lymphocyte cell viability and proliferation in concentrations ranging from "safe" to "retinotoxic" in relation to intravitreal injection volumes. Low-dose intravitreal gentamicin may not suppress the Th1- and Th17-mediated immune response.
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Enfermedades de los Caballos , Uveítis , Animales , Caballos , Gentamicinas/farmacología , Gentamicinas/uso terapéutico , Proyectos de Investigación , Uveítis/tratamiento farmacológico , Uveítis/veterinaria , Linfocitos T , Proliferación Celular , Enfermedades de los Caballos/tratamiento farmacológicoRESUMEN
PURPOSE: To evaluate the normal refractive state in horses in NCSU and ECMR and determine the prevalence of naturally occurring refractive errors and their association with breed, age, coat color, iris color, sex, and geographic location. METHODS: Horses from NCSU (January 2009-November 2012) and ECMR (January 2013-September 2016) underwent ophthalmic examination and streak retinoscopy. Location, color, breed, sex, and iris color were recorded. Gross and net refractive values for each meridian (horizontal and vertical), spherical refraction, astigmatism for both eyes, and anisometry were recorded, and statistical analyses were performed. RESULTS: There is excellent agreement in refraction between the eyes of the same horse (ICC = 0.89). The median net horizontal (H), vertical (V), and spherical refraction for the total population (n = 690) were H: +0.25 D (min. -6.50 D, max. +2.34 D), V: +0.25 D (min. -7.13 D, max. +2.75D), and spherical: +0.25 D (min. -6.82 D, max. +2.17 D), all with interquartile ranges of -0.25 to 0.25 D. Emmetropia (>-0.50 D and <+0.50 D; >-0.75 D and <+0.75 D) was present in 769/1380 eyes (55.7%) and 926/1380 eyes (67.1%), respectively. Anisometropia was present in 86/690 horses (12.5%). Sex, iris color, and location were significantly associated with refraction values, whereas age, breed, and coat color were not. CONCLUSIONS: Most eyes evaluated are emmetropic, or shifted myopically, with excellent agreement between eyes of the same horse. Sex, iris color, and geographic location appear to impact refraction in horses. SUPPORT: None.
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Enfermedades de los Caballos , Errores de Refracción , Caballos , Animales , Prevalencia , North Carolina , Errores de Refracción/epidemiología , Errores de Refracción/veterinaria , Refracción Ocular , Alemania/epidemiología , IrisRESUMEN
In this article, which is part of the Currents in One Health series, the role of naturally occurring ocular disease in animals is reviewed with emphasis on how the understanding of these ocular diseases contributes to one health initiatives, particularly the pathogenesis and treatment of ocular diseases common to animals and humans. Animals spontaneously develop ocular diseases that closely mimic those in humans, especially dry eye disease, herpes virus infection (cats), fungal keratitis (horses), bacterial keratoconjunctivitis, uveitis, and glaucoma. Both uveitis and glaucoma are common in domestic animals and humans, and many similarities exist in pathogenesis, genetics, and response to therapy. Furthermore, the study of inherited retinal disease in animals has particularly epitomized the one health concept, specifically the collaborative efforts of multiple disciplines working to attain optimal health for people and animals. Through this study of retinal disease in dogs, innovative therapies such as gene therapy have been developed. A unique opportunity exists to study ocular disease in shared environments to better understand the interplay between the environment, genetics, and ocular disease in both animals and humans. The companion Currents in One Health by Gilger, AJVR, December 2022, addresses in more detail recent studies of noninfectious immune-mediated animal ocular disease and their role in advancing ocular health globally.
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Enfermedades de los Gatos , Úlcera de la Córnea , Enfermedades de los Perros , Glaucoma , Enfermedades de los Caballos , Enfermedades de la Retina , Uveítis , Humanos , Caballos , Gatos , Animales , Perros , Ojo , Uveítis/epidemiología , Uveítis/terapia , Uveítis/veterinaria , Glaucoma/veterinaria , Enfermedades de la Retina/veterinaria , Úlcera de la Córnea/veterinaria , Enfermedades de los Perros/epidemiología , Enfermedades de los Perros/terapia , Enfermedades de los Caballos/patologíaRESUMEN
This review, which is part of the "Currents in One Health" series, describes the importance of the study of immune-mediated ocular disease in the development of innovative therapeutics, such as cell and gene therapy for the eye. Recent examples of cell and gene therapy studies from the author's laboratory are reviewed to emphasize the importance of One Health initiatives in developing innovative therapies for ocular diseases. Spontaneous immune-mediated corneal disease is common in horses, cats, dogs, and humans. Autologous bone marrow-derived mesenchymal stem cells (BM-MSCs) injected subconjunctivally resulted in the resolution of naturally occurring immune-mediated keratitis (IMMK) without adverse effects. These results support that autologous subconjunctival BM-MSC therapy may be a viable treatment alternative for IMMK. Furthermore, the use of subconjunctival MSCs may be an effective method to treat ocular surface immune-mediated diseases in humans and other species, including herpetic stromal keratitis and immunologic dry eye disease. Furthermore, the use of adeno-associated viral (AAV) vectors to deliver the immunosuppressive transgene cDNA of equine interleukin 10 (eqIL-10) or human leukocyte antigen G injected intravitreally was shown to be safe and inhibited the development of uveitis in the experimental autoimmune uveitis rat model. Efficacy and safety studies of ocular gene therapy in models will pave the way for clinical trials in animals with naturally occurring immune-mediated diseases, such as a therapeutic clinical trial for AAV-eqIL-10 in horses with equine recurrent uveitis.
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Enfermedades de los Gatos , Enfermedades de los Perros , Enfermedades de los Caballos , Queratitis , Células Madre Mesenquimatosas , Enfermedades de los Roedores , Uveítis , Caballos , Animales , Humanos , Ratas , Gatos , Perros , Animales Domésticos , Ojo , Queratitis/metabolismo , Queratitis/veterinaria , Uveítis/terapia , Uveítis/veterinaria , Enfermedades de los Perros/metabolismo , Enfermedades de los Caballos/terapia , Enfermedades de los Caballos/metabolismo , Enfermedades de los Roedores/metabolismoRESUMEN
Equine recurrent uveitis (ERU) is a spontaneous, painful, and vision threatening disease affecting up to 25% of equine populations worldwide. Current treatments of ERU are non-specific and have many side effects which limits them to short-term use. In order to develop an effective therapy for ERU, we investigated the use of adeno-associated virus (AAV) gene therapy, exploiting a natural immune tolerance mechanism induced by equine interleukin-10 (Equine-IL10). The purpose of this study was to evaluate the therapeutic efficacy of a single intravitreal (IVT) dose of AAV8-Equine-IL10 gene therapy for inhibition of experimental autoimmune uveitis (EAU) in rats. Each rat was dosed intravitreally (IVT) in both eyes with either balanced salt solution (BSS) (control; n = 4), AAV8-Equine-IL10 at a low dose (2.4x109 vg; n = 5) or high dose (2.4x1010 vg; n = 5). EAU was induced in all groups of rats 7 days after IVT injections and euthanized 21 days post-injection. Ophthalmic examination and aqueous humor (AH) cell counts were recorded with the observer blinded to the treatment groups. Histopathology and qPCR were performed on selected ocular tissues. Data presented herein demonstrate that AAV8-Equine-IL10 treated rats exhibited a significant decrease in clinical inflammatory scores and AH cell counts compared to BSS-treated EAU eyes on days 10, 12 and 14 post EAU induction at both administered vector doses. Mean cellular histologic infiltrative scores were also significantly less in AAV8-Equine-IL10 dosed rats compared to the BSS group. Intravitreal injection of AAV8-Equine-IL10 resulted in Equine-IL10 cDNA expression in the ciliary body, retina, cornea, and optic nerve in a dose-dependent manner. A single IVT injection of AAV8-Equine-IL10 appeared to be well-tolerated and inhibited EAU even at the lowest administered dose. These results demonstrate safety and efficacy of AAV8-Equine-IL10 to prevent EAU and support continued exploration of AAV gene therapy for the treatment of equine and perhaps human recurrent uveitis.
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Enfermedades Autoinmunes , Uveítis , Animales , Dependovirus/genética , Terapia Genética , Caballos/genética , Humanos , Interleucina-10/genética , Interleucina-10/uso terapéutico , RatasRESUMEN
Deficiency and dysfunction of corneal cells leads to the blindness observed in corneal diseases such as limbal stem cell deficiency (LSCD) and bullous keratopathy. Regenerative cell therapies and engineered corneal tissue are promising treatments for these diseases [1]. However, these treatments are not yet clinically feasible due to inadequate cell sources. The discovery of induced pluripotent stem cells (iPSCs) by Shinya Yamanaka has provided a multitude of opportunities in research because iPSCs can be generated from somatic cells, thus providing an autologous and unlimited source for corneal cells. Compared to other stem cell sources such as mesenchymal and embryonic, iPSCs have advantages in differentiation potential and ethical concerns, respectively. Efforts have been made to use iPSCs to model corneal disorders and diseases, drug testing [2], and regenerative medicine [1]. Autologous treatments based on iPSCs can be exorbitantly expensive and time-consuming, but development of stem cell banks with human leukocyte antigen (HLA)- homozygous cell lines can provide cost- and time-efficient allogeneic alternatives. In this review, we discuss the early development of the cornea because protocols differentiating iPSCs toward corneal lineages rely heavily upon recapitulating this development. Differentiation of iPSCs toward corneal cell phenotypes have been analyzed with an emphasis on feeder-free, xeno-free, and well-defined protocols, which have clinical relevance. The application, challenges, and potential of iPSCs in corneal research are also discussed with a focus on hurdles that prevent clinical translation.
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Enfermedades de la Córnea , Células Madre Pluripotentes Inducidas , Humanos , Diferenciación Celular/genética , Córnea , Línea Celular , Enfermedades de la Córnea/terapiaRESUMEN
OBJECTIVE: To describe the use of 1% polidocanol as the sole treatment for a superficial orbital venous malformation in a horse. ANIMAL: A 23-year-old Welsh Cobb cross gelding with a distensible swelling affecting the left lower eyelid, and secondary palpebral margin abnormalities and superficial keratitis. PROCEDURE: Color flow Doppler ultrasonography revealed non-pulsatile blood flow within the tortuous vascular network most consistent with a superficial orbital venous malformation appearing to involve the lateral palpebral and transverse facial veins. An intravenous catheter was placed within the lateral aspect of the venous malformation, and agitated saline was slowly injected into the vessel while simultaneously ultrasound imaging the medial aspect in which the bubbles were observed coursing across, consistent with lateral to medial flow. Contrast venography confirmed a corkscrew vessel along the ventral aspect of the orbital rim. Under standing sedation, 1% polidocanol solution was administered slowly through the intravenous catheter while manual pressure was applied on the medial and lateral aspects of the venous malformation. RESULTS: Ultrasonography performed immediately after administration of polidocanol confirmed venous stasis, and formation of a thrombus. No adverse side effects were noted. The venous malformation and associated palpebral margin abnormalities and superficial keratitis resolved at the time of re-examination at 4 months. CONCLUSION AND CLINICAL RELEVANCE: Polidocanol as the sole treatment for a superficial orbital venous malformation in a horse was well tolerated and led to clinical resolution. Sclerosant monotherapy may be a safe treatment option for superficial orbital venous malformations.
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Enfermedades de los Caballos , Enfermedades Orbitales , Animales , Enfermedades de los Caballos/inducido químicamente , Enfermedades de los Caballos/diagnóstico por imagen , Enfermedades de los Caballos/tratamiento farmacológico , Caballos , Masculino , Enfermedades Orbitales/veterinaria , Polidocanol/uso terapéutico , Polietilenglicoles , Soluciones Esclerosantes/efectos adversos , Soluciones Esclerosantes/uso terapéutico , Escleroterapia/veterinaria , Resultado del Tratamiento , Venas/anomalíasRESUMEN
The purpose of this paper is to review human leukocyte antigen G (HLA-G) in the eye, its role in immune tolerance, and the potential therapeutic use of AAV gene transfer and expression of HLA-G in various ocular tissues. Several studies are reviewed that demonstrate efficacy in animal models of disease, including intracorneal delivery of AAV-HLA-G to treat corneal inflammation and prevent corneal graft rejection, subconjunctival injection of AAV-HLA-G for ocular graft vs. host disease and potentially dry eye disease, and intravitreal injection of AAV-HLA-G to inhibit uveitis. Furthermore, due to the anti-vascular function of HLA-G, AAV-HLA-G may be an effective therapy for posterior ocular diseases, such as neovascular age-related macular degeneration, diabetic retinopathy, and choroidal neovascularization. Therefore, AAV-mediated gene transfer of HLA-G may be an effective treatment for common immune-mediated, inflammatory, and neovascular diseases of the eye.
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Neovascularización Coroidal , Dependovirus , Animales , Neovascularización Coroidal/genética , Dependovirus/genética , Terapia Genética , Vectores Genéticos/genética , Antígenos HLA-G/genéticaRESUMEN
Purpose: To determine if inhibition of Myristoylated Alanine Rich C Kinase Substrate (MARCKS) protein, using novel MARCKS inhibitor peptides, will reduce the severity of endotoxin-induced uveitis (EIU) in rats. Methods: EIU was induced in Lewis rats using subcutaneous administration of lipopolysaccharide. In the first phase of the study, 3 different novel MARCKS inhibitor peptides that mimic the N-terminal region of MARCKS (BIO-11006, or lower molecular weight analogs BIO-91201 or BIO-91202; Biomarck Pharmaceuticals, Ltd., Newtown, PA) were administered intravitreally (IVT) at 50 and 100 µM. In the second phase, BIO-91201 was administered IVT at 10, 50, and 100 µM and topically at the 100 µM concentration. The efficacy of MARCKS inhibitor peptides was assessed by clinical examination using slit lamp biomicroscopy, optical coherence tomography (OCT) anterior chamber cell counts, histopathology, and aqueous humor cytokine analysis. Results: Clinical scores were significantly reduced 24 h following uveitis induction in the first phase of the study in the following treatment groups: BIO-11006 50 µM IVT and 100 µM IVT, BIO-91201 50 µM IVT, and BIO-91202 100 µM IVT (P < 0.05). OCT anterior chamber cell counts were significantly reduced in the first phase of the study in all treatment groups (P < 0.001). OCT anterior chamber cell counts and histopathology scores were significantly reduced in the second phase of the study in the BIO-91201 50 µM IVT group (P < 0.05). No effect was seen with topical administration. Conclusion: MARCKS inhibitor peptides were effective in reducing the severity of ocular inflammation and cellular influx in EIU.
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Endotoxinas , Uveítis , Animales , Humor Acuoso/metabolismo , Endotoxinas/toxicidad , Lipopolisacáridos/metabolismo , Lipopolisacáridos/farmacología , Sustrato de la Proteína Quinasa C Rico en Alanina Miristoilada/metabolismo , Péptidos/farmacología , Péptidos/uso terapéutico , Ratas , Ratas Endogámicas Lew , Uveítis/inducido químicamente , Uveítis/tratamiento farmacológico , Uveítis/patologíaRESUMEN
BACKGROUND: Superficial chronic corneal epithelial defects (SCCEDs) are spontaneous corneal defects in dogs that share many clinical and pathologic characteristics to recurrent corneal erosions (RCE) in humans. Boxer dogs are predisposed to SCCEDs, therefore a search for a genetic defect was performed to explain this susceptibility. DNA was extracted from blood collected from Boxer dogs with and without SCCEDs followed by whole genome sequencing (WGS). RNA sequencing of corneal tissue and immunostaining of corneal sections from affected SCCED Boxer dogs with a deletion in the NOG gene and affected non-Boxer dogs without the deletion were performed. RESULTS: A 30 base pair deletion at a splice site in Noggin (NOG) (Chr 9:31453999) was identified by WGS and was significantly associated (P < 0.0001) with Boxer SCCEDs compared to unaffected non-Boxer dogs. NOG, BMP4, MMP13, and NCAM1 all had significant fold reductions in expression and SHH was significantly increased in Boxers with the NOG deletion as identified by RNA-Seq. Corneal IHC from NOG deletion dogs with SCCEDs had lower NOG and significantly higher scores of BMP2. CONCLUSIONS: Many Boxer dogs with SCCED have a genetic defect in NOG. NOG is a constitutive protein in the cornea which is a potent inhibitor of BMP, which likely regulate limbal epithelial progenitor cells (LEPC). Dysregulation of LEPC may play a role in the pathogenesis of RCE.
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Proteínas Morfogenéticas Óseas/metabolismo , Enfermedades de los Perros/genética , Predisposición Genética a la Enfermedad , Animales , Proteínas Morfogenéticas Óseas/genética , Enfermedad Crónica , Enfermedades de la Córnea/veterinaria , Enfermedades de los Perros/patología , Perros , Epitelio Corneal/patología , Regulación de la Expresión Génica , Secuenciación Completa del GenomaRESUMEN
OBJECTIVE: Equine recurrent uveitis (ERU), a chronic, immune-mediated intraocular inflammatory disease, is a common cause of blindness in horses. The severity and recurrent nature of ERU makes it difficult to treat with current therapeutics leading to a poor visual prognosis. The suprachoroidal space (SCS), a potential space between the choroid and sclera surrounding the ocular posterior segment, offers a promising alternative site for drug application to the eye. Corticosteroid administration within this space is hypothesized to be safe and effective at controlling intraocular inflammation, especially in horses with poorly responsive ERU. ANIMAL STUDIED: Horses with active, poorly responsive ERU. PROCEDURE(S): A retrospective study was performed with 29 horses (36 total eyes) that received SCS injection of triamcinolone acetonide (TA) with ERU not well controlled with standard uveitis treatment. A standardized ocular inflammation score (OIS) was used to assess inflammation at the time of injection and at follow-ups. RESULTS: Standardized OIS revealed a significant decrease in ocular inflammation over time after SCS TA administration (p < .004). Adverse effects after injections occurred in <20% of the horses at follow-up, but some of these effects were attributed to chronic inflammation prior to effective treatment, long-term topical corticosteroid use, or complications from hospitalization rather than the SCS injections. Most horses (86.7%) in this study remained visual greater than 3 months after SCS injection. CONCLUSIONS: Based on these results, SCS TA injections appear to be a safe and possible effective treatment modality for managing poorly responsive ERU; further clinical study is warranted.