RESUMO
Bietti crystalline corneoretinal dystrophy is an inherited retinal disease caused by mutations in CYP4V2, which results in blindness in the working-age population, and there is currently no available treatment. Here, we report the results of the first-in-human clinical trial (NCT04722107) of gene therapy for Bietti crystalline corneoretinal dystrophy, including 12 participants who were followed up for 180-365 days. This open-label, single-arm exploratory trial aimed to assess the safety and efficacy of a recombinant adeno-associated-virus-serotype-2/8 vector encoding the human CYP4V2 protein (rAAV2/8-hCYP4V2). Participants received a single unilateral subretinal injection of 7.5 × 1010 vector genomes of rAAV2/8-hCYP4V2. Overall, 73 treatment-emergent adverse events were reported, with the majority (98.6%) being of mild or moderate intensity and considered to be procedure- or corticosteroid-related; no treatment-related serious adverse events or local/systemic immune toxicities were observed. Compared with that measured at baseline, 77.8% of the treated eyes showed improvement in best-corrected visual acuity (BCVA) on day 180, with a mean ± standard deviation increase of 9.0 ± 10.8 letters in the 9 eyes analyzed (p = 0.021). By day 365, 80% of the treated eyes showed an increase in BCVA, with a mean increase of 11.0 ± 10.6 letters in the 5 eyes assessed (p = 0.125). Importantly, the patients' improvement observed using multifocal electroretinogram, microperimetry, and Visual Function Questionnaire-25 further supported the beneficial effects of the treatment. We conclude that the favorable safety profile and visual improvements identified in this trial encourage the continued development of rAAV2/8-hCYP4V2 (named ZVS101e).
Assuntos
Distrofias Hereditárias da Córnea , Família 4 do Citocromo P450 , Dependovirus , Terapia Genética , Doenças Retinianas , Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Adulto , Distrofias Hereditárias da Córnea/genética , Distrofias Hereditárias da Córnea/terapia , Distrofias Hereditárias da Córnea/patologia , Dependovirus/genética , Família 4 do Citocromo P450/genética , Vetores Genéticos/genética , Acuidade VisualRESUMO
PURPOSE: The purpose of this article was to describe the successful diagnosis and management of clinically atypical, unilateral, gelatinous drop-like corneal dystrophy (GDLD) in a pediatric patient. METHODS: This study was a case report. RESULTS: A 7-year-old Japanese girl was referred to our clinic with right corneal opacity for over 3 years. Slitlamp examination revealed a white, protruding, paracentral corneal opacity with an irregular surface and tiny stromal lattice figures with subepithelial opacities. No trichiasis or epiblepharon was observed, and the patient's right corrected distance visual acuity (CDVA) was 18/20. The contralateral cornea was intact but demonstrated fluorescein uptake. After 8 months, the right CDVA worsened from 18/20 to 6/20, and corneal epithelial scraping was performed. Histopathological analysis revealed amyloid nodules in the subepithelial layer and in the anterior corneal stroma stained with Congo red, which reoccurred 2 months after the procedure, and corneal dystrophy was suspected. Isolation and sequencing of the genomic DNA revealed a homozygous p.Gln118Ter. mutation in TACSTD2 in the patient and heterozygous p.Gln118Ter. mutations in both parents. GDLD was diagnosed; bilateral use of therapeutic soft contact lenses was prescribed after the first corneal scraping. No additional surgical intervention was required for the right eye for 4.5 years. CDVA of the contralateral left eye has been successfully maintained at 30/20 over this period, without emergence of nodular lesions or corneal opacities. CONCLUSIONS: We encountered a patient with early, atypical GDLD, who was definitively diagnosed using genomic DNA sequencing. GDLD should be a part of the differential diagnosis in patients presenting with unilateral, recurrent amyloid deposition.
Assuntos
Distrofias Hereditárias da Córnea , Opacidade da Córnea , Amiloidose Familiar , Antígenos de Neoplasias/genética , Moléculas de Adesão Celular/genética , Criança , Vermelho Congo , Distrofias Hereditárias da Córnea/diagnóstico , Distrofias Hereditárias da Córnea/genética , Distrofias Hereditárias da Córnea/patologia , Opacidade da Córnea/diagnóstico , Opacidade da Córnea/genética , DNA/genética , Feminino , Fluoresceínas , Gelatina , HumanosRESUMO
Corneal dystrophies describe a clinically and genetically heterogeneous group of inherited disorders. The International Classification of Corneal Dystrophies (IC3D) lists 22 types of corneal dystrophy, 17 of which have been demonstrated to result from pathogenic variants in 19 identified genes. In this study, we investigated the diagnostic yield of genetic testing in a well-characterised cohort of 58 individuals from 44 families with different types of corneal dystrophy. Individuals diagnosed solely with Fuchs endothelial corneal dystrophy were excluded. Clinical details were obtained from the treating ophthalmologist. Participants and their family members were tested using a gene candidate and exome sequencing approach. We identified a likely molecular diagnosis in 70.5% families (31/44). The detection rate was significantly higher among probands with a family history of corneal dystrophy (15/16, 93.8%) than those without (16/28, 57.1%, p = .015), and among those who had undergone corneal graft surgery (9/9, 100.0%) compared to those who had not (22/35, 62.9%, p = .041). We identified eight novel variants in five genes and identified five families with syndromes associated with corneal dystrophies. Our findings highlight the genetic heterogeneity of corneal dystrophies and the clinical utility of genetic testing in reaching an accurate clinical diagnosis.
Assuntos
Distrofias Hereditárias da Córnea , Austrália , Estudos de Coortes , Distrofias Hereditárias da Córnea/diagnóstico , Distrofias Hereditárias da Córnea/genética , Distrofias Hereditárias da Córnea/patologia , Testes Genéticos , HumanosRESUMO
Material can be deposited in the cornea as a result of a wide range of systemic and ophthalmic diseases, as well as local and systemic therapies. Causes include local infection or trauma, systemic malignancy, a wide range of medications and a host of genetic and metabolic diseases. Some of these can be acutely life threatening, so generalists caring for both children and adults should have a basic awareness of the pattern and distribution of corneal deposits to facilitate timely diagnosis, investigation, management or onward referral to avoid significant morbidity or mortality. This article outlines causes of corneal deposits found in patients presenting to primary care, ophthalmic clinics or encountered on the wards to help generalists avoid missing serious pathology. It also provides insight into the natural history of underlying causative conditions and their possible treatments.
Assuntos
Distrofias Hereditárias da Córnea , Adulto , Criança , Córnea/patologia , Distrofias Hereditárias da Córnea/diagnóstico , Distrofias Hereditárias da Córnea/genética , Distrofias Hereditárias da Córnea/patologia , HumanosRESUMO
BACKGROUND: Macular corneal dystrophy is a rare inherited disease of the cornea leading to deposits mainly in the stroma. Affected patients suffer from progressive loss of visual acuity which should be treated with penetrating keratoplasty. This is the first case report describing the clinical and histopathological findings of corneal tissue after failed phototherapeutic keratectomy (PTK) in a patient with macular corneal dystrophy. CASE PRESENTATION: A 32-year-old man presented with visual impairment, blurred vision and increasing glare sensitivity in both eyes in 2014. All symptoms had existed for several years and had recently increased sharply. A corneal dystrophy was diagnosed and penetrating keratoplasty was recommended but the patient was hesitant to undergo surgery. In 2018, in contrast to current guidelines, a PTK was performed in both eyes in Turkey for unknown reasons. In May 2019, he presented again in our clinic. Best corrected visual acuity was markedly reduced in both eyes. Slit-lamp examination revealed multiple dense, poorly circumscribed grey-white patchy changes in the stroma accompanied by corneal opacity in both eyes. In February 2020, the patient decided to have penetrating keratoplasty performed at the University Eye Hospital in Tübingen. The explanted cornea was stained for acid mucopolysaccharides (AMP) and periodic acid-Schiff staining (PAS). The histopathological examination revealed destruction of Bowman's layer and a subepithelial fibrosis band due to the PTK previously performed. The AMP staining demonstrated blue deposits typical of macular corneal dystrophy, mainly in the stroma but also in the endothelium. Interestingly, the acidic mucopolysaccharides were found increased in the PTK-induced subepithelial fibrosis band. The postoperative course after keratoplasty was favourable with a significant increase in visual acuity and a clear graft. CONCLUSIONS: This report presents the first case of a histologically evident exacerbation of macular corneal dystrophy after PTK and emphasizes the relevance of thorough pre-interventional diagnosis and patient selection to consider other therapeutic approaches, such as penetrating keratoplasty.
Assuntos
Distrofias Hereditárias da Córnea , Ceratectomia Fotorrefrativa , Adulto , Humanos , Masculino , Córnea/patologia , Córnea/cirurgia , Distrofias Hereditárias da Córnea/diagnóstico , Distrofias Hereditárias da Córnea/patologia , Distrofias Hereditárias da Córnea/cirurgia , Fibrose , Seguimentos , Glicosaminoglicanos , Ceratectomia , Lasers de ExcimerRESUMO
Thiel-Behnke corneal dystrophy (TBCD) is an epithelial-stromal TGFBI dystrophy caused by mutations in the TGFBI (transforming growth factor beta induced) gene, though the underlying mechanisms and pathogenesis of TBCD are still obscure. The study identifies a novel mutation in the TGFBI gene (p.Gly623_His626del) in a TBCD pedigree. Characteristics of the typical vacuole formation, irregular corneal epithelial thickening and thinning, deposition of eosinophilic substances beneath the epithelium, and involvement of the anterior stroma were observed in this pedigree via transmission electron microscopy (TEM) and histological staining. Tgfbi-p.Gly623_Tyr626del mouse models of TBCD were subsequently generated via CRISPR/Cas9 technology, and the above characteristics were further verified via TEM and histological staining. Lysosomal dysfunction and downregulation of differential expression protein CTSD (cathepsin D) were observed using LysoTracker Green DND-26 and proteomic analysis, respectively. Hence, lysosomal dysfunction probably leads to autophagic flux obstruction in TBCD; this was supported by enhanced LC3-II and SQSTM1 levels and decreased CTSD. TFEB (transcription factor EB) was prominently decreased in TBCD corneal fibroblasts and administration of ATP-competitive MTOR inhibitor torin 1 reversed this decline, resulting in the degradation of accumulated mut-TGFBI (mutant TGFBI protein) via the ameliorative lysosomal function and autophagic flux owing to elevated TFEB activity as measured by western blot, confocal microscopy, and flow cytometry. Transfected HEK 293 cells overexpressing human full-length WT-TGFBI and mut-TGFBI were generated to further verify the results obtained in human corneal fibroblasts. Amelioration of lysosome dysfunction may therefore have therapeutic efficacy in the treatment of TBCD.Abbreviations AS-OCT: anterior segment optical coherence tomography; ATP: adenosine triphosphate; Cas9: CRISPR-associated protein 9; CLEAR: coordinated lysosomal expression and regulation; CRISPR: clustered regularly interspaced short palindromic repeats; CTSB: cathepsin B; CTSD: cathepsin D; CTSF: cathepsin F; CTSL: cathepsin L; DNA: deoxyribonucleic acid; ECM: extracellular matrix; Fas1: fasciclin 1; FC: flow cytometry; GAPDH: glyceraldeyde-3-phosphate dehydrogenase; GCD2: granular corneal dystrophy type 2; HE: hematoxylin and eosin; LAMP2: lysosomal-associated membrane protein; MT: mutation type; MTOR: mechanistic target of rapamycin kinase; MTORC1: MTOR complex 1; mut-TGFBI: mutant TGFBI protein; SD: standard deviation; TBCD: Thiel-Behnke corneal dystrophy; TEM: transmission electron microscopy; TFEB: transcription factor EB; TGFBI: transforming growth factor beta induced; WT: wild type.
Assuntos
Catepsina D , Distrofias Hereditárias da Córnea , Trifosfato de Adenosina/metabolismo , Animais , Autofagia/genética , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos , Proteínas Sanguíneas , Catepsina D/metabolismo , Distrofias Hereditárias da Córnea/genética , Distrofias Hereditárias da Córnea/metabolismo , Distrofias Hereditárias da Córnea/patologia , Células HEK293 , Humanos , Lisossomos/metabolismo , Camundongos , Proteínas Associadas aos Microtúbulos/metabolismo , Proteínas Mutantes/metabolismo , Proteômica , Serina-Treonina Quinases TOR/metabolismo , Fator de Crescimento Transformador beta/metabolismoRESUMO
A 17-year-old girl with a few years' history of declining vision, photophobia, and dry eye symptoms was referred to our clinic. She noted that the vision in the right eye declined significantly over the past several months. On her last year examination, her uncorrected distance visual acuity (UDVA) was recorded as 20/25 in both eyes with a corrected distance visual acuity (CDVA) of 20/20 in both eyes with minimal refractive error, with a diagnosis of bilateral Salzmann nodular degeneration. The patient was given artificial tears and was encouraged to wear sunglasses. On examination now, UDVA was 20/70 in the right eye and 20/40 in the left eye. The manifest refraction was -2.00 + 1.25 × 96 in the right eye and -1.00 + 2.00× 34 in the left eye, with a CDVA of 20/50 and 20/30, respectively. Slitlamp examination revealed superficial reticular stromal scar with clear intervening spaces involving the anterior 75 µm of the stromal cornea in the central 6.0 mm optical zone (Figure 1).JOURNAL/jcrs/04.03/02158034-202104000-00021/figure1/v/2021-04-19T183640Z/r/image-tiffJOURNAL/jcrs/04.03/02158034-202104000-00021/figure2/v/2021-04-19T183640Z/r/image-tiff The rest of the anterior and posterior segment examination was completely normal and noncontributory. Anterior segment optical coherence tomography (AS-OCT) revealed subepithelial lesion involving the central aspect of the cornea in the right eye more than that in the left eye (Figure 2). Family history was significant for an older sister with a similar problem who never required medical attention. She also has mild photophobia and dry eye symptoms. What is your differential diagnosis? What diagnostic test will help you in your diagnosis and clinical decision-making? What is the most likely diagnosis in this case? Do you recommend medical and/or surgical intervention in the right eye, realizing that there has been exacerbation of her ocular condition in the most recent year? What is the long-term prognosis and future plan for a patient with this potential condition?
Assuntos
Cicatriz , Distrofias Hereditárias da Córnea , Adolescente , Cicatriz/patologia , Córnea/patologia , Distrofias Hereditárias da Córnea/patologia , Feminino , Humanos , Tomografia de Coerência Óptica , Acuidade VisualRESUMO
Harboyan syndrome or corneal dystrophy and progressive deafness (MIM #217400) is characterized by congenital hereditary endothelial dystrophy (CHED) and progressive, sensorineural hearing loss. Mutations in SLC4A11 are responsible for this rare genetic syndrome. Eight patients from seven unrelated families affected with Harboyan Syndrome with mean follow-up of 12.0 ± 0.9 years were thoroughly investigated for the ocular, hearing, and kidney function abnormalities and the outcome of penetrating keratoplasty (PK). Mutation analysis of SLC4A11 was performed. All patients presented with bilateral cloudy corneas since birth. Sensorineural hearing loss was detected in all patients. Seven patients (11 eyes) underwent PK with the median age at surgery of 10.1 years (7.1-22.9). The overall corneal graft survival rate after primary PK was 72.7% (8/11 eyes). The mean graft survival time was 94.6 months (95% CI 83.1-126.0). All patients had unremarkable kidney function. The c.2264G>A (p.Arg755Gln) mutation in SCL4A11 was detected in most patients (87.5%). All unrelated Karen tribe patients had p.Arg755Gln mutation, suggestive of founder effect. We found the allele frequency of this variant in the Karen population to be 0.01. The c.2263C>T (p.Arg755Trp) mutation was found in one patient with mild phenotype and the novel truncating protein mutation c.2127delG (p.Gly710fsx*25) in SCL4A11 was identified in two Thai sisters. Visual outcome and graft survival after PK were satisfactory. Our study shows that all studied patients with SLC4A11 mutations had CHED and sensorineural hearing loss, and SLC4A11 mutations were not related to the onset and severity of hearing loss or outcome of keratoplasty.
Assuntos
Proteínas de Transporte de Ânions/genética , Antiporters/genética , Distrofias Hereditárias da Córnea/genética , Distrofias Hereditárias da Córnea/patologia , Transplante de Córnea/métodos , Perda Auditiva Neurossensorial/genética , Perda Auditiva Neurossensorial/patologia , Mutação , Fenótipo , Adolescente , Adulto , Proteínas de Transporte de Ânions/química , Antiporters/química , Criança , Pré-Escolar , Distrofias Hereditárias da Córnea/cirurgia , Feminino , Efeito Fundador , Perda Auditiva Neurossensorial/cirurgia , Humanos , Lactente , Masculino , Linhagem , Conformação Proteica , Adulto JovemRESUMO
Gelatinous drop-like corneal dystrophy (GDLD) is a severe inherited corneal dystrophy characterized by subepithelial corneal amyloid deposition. We had previously succeeded in identifying the responsible gene, TACSTD2, and subsequently found that the epithelial barrier function is significantly decreased. As with GDLD patients, the knockout mice showed severe loss of tight junction, progressive opacity, and neovascularization in the cornea. We devised an easy method to confirm the loss of the corneal barrier function even before corneal opacity is observed. Furthermore, by using knockout mice, we were able to verify clinical findings, such as the wound healing delay and light-induced acceleration of the disease. This mouse model should prove to be a highly useful tool for investigating the pathology of GDLD and for developing new therapies.
Assuntos
Amiloidose Familiar/patologia , Antígenos de Neoplasias/genética , Moléculas de Adesão Celular/genética , Distrofias Hereditárias da Córnea/patologia , Animais , Distrofias Hereditárias da Córnea/genética , Modelos Animais de Doenças , Gelatina/genética , Gelatina/metabolismo , Camundongos , Mutação/genéticaRESUMO
Neovascular retinal degeneration is a leading cause of blindness in advanced countries. Anti-vascular endothelial growth factor (VEGF) drugs have been used for neovascular retinal diseases; however, anti-VEGF drugs may cause the development of chorioretinal atrophy in chronic therapy as they affect the physiological amount of VEGF needed for retinal homeostasis. Hypoxia-inducible factor (HIF) is a transcription factor inducing VEGF expression under hypoxic and other stress conditions. Previously, we demonstrated that HIF was involved with pathological retinal angiogenesis in murine models of oxygen-induced retinopathy (OIR), and pharmacological HIF inhibition prevented retinal neovascularization by reducing an ectopic amount of VEGF. Along with this, we attempted to find novel effective HIF inhibitors. Compounds originally isolated from mushroom-forming fungi were screened for prospective HIF inhibitors utilizing cell lines of 3T3, ARPE-19 and 661W. A murine OIR model was used to examine the anti-angiogenic effects of the compounds. As a result, 2-azahypoxanthine (AHX) showed an inhibitory effect on HIF activation and suppressed Vegf mRNA upregulation under CoCl2-induced pseudo-hypoxic conditions. Oral administration of AHX significantly suppressed retinal neovascular tufts in the OIR model. These data suggest that AHX could be a promising anti-angiogenic agent in retinal neovascularization by inhibiting HIF activation.
Assuntos
Inibidores da Angiogênese/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Fator 1 Induzível por Hipóxia/antagonistas & inibidores , Neovascularização Retiniana/metabolismo , Fator A de Crescimento do Endotélio Vascular/biossíntese , Células 3T3 , Inibidores da Angiogênese/química , Animais , Hipóxia Celular/efeitos dos fármacos , Cobalto/toxicidade , Distrofias Hereditárias da Córnea/induzido quimicamente , Distrofias Hereditárias da Córnea/metabolismo , Distrofias Hereditárias da Córnea/patologia , Modelos Animais de Doenças , Fator 1 Induzível por Hipóxia/metabolismo , Camundongos , Neovascularização Retiniana/induzido quimicamente , Neovascularização Retiniana/patologiaRESUMO
BACKGROUND: Most transforming growth factor beta-induced (TGFBI) corneal dystrophies are associated with a characteristic phenotype, clinical course, and a conserved mutation in the TGFBI gene. However, we report a novel TGFBI missense mutation associated with a late-onset, variant Bowman layer dystrophy. METHODS: Participants underwent slit-lamp examination and multimodal imaging. Polymerase chain reaction amplification and Sanger sequencing were performed on saliva-derived genomic DNA to screen TGFBI exons 4 and 12 as well as COL17A1 exon 46. PolyPhen-2 and SIFT were used to predict the functional impact of any identified variants. RESULTS: A 56-year-old Thai woman reported a four-year history of decreased vision and intermittent eye irritation, suggestive of recurrent epithelial erosions, in both eyes. Slit-lamp exam revealed bilateral, irregular, limbal-sparing Bowman layer opacities, which were also noted on anterior segment optical coherence tomography. Phototherapeutic keratectomy was performed in the right eye, improving the best-corrected visual acuity from 20/50 to 20/30. Sequencing of the TGFBI gene revealed a novel heterozygous, missense mutation in exon 12 (c.1571 C > G; p.Ser524Cys), which was present in an affected son and absent in an unaffected son, and was predicted to be damaging by PolyPhen-2 and SIFT. The patient was diagnosed with a variant Bowman layer dystrophy given the late onset of an atypical phenotype and the identification of a novel TGFBI mutation. CONCLUSIONS: A novel TGFBI missense mutation is associated with a late-onset Bowman layer dystrophy. Given the atypical clinical appearance and course, molecular genetic analysis was utilized to establish a definitive diagnosis.
Assuntos
Lâmina Limitante Anterior/patologia , Distrofias Hereditárias da Córnea/patologia , Epitélio Corneano/patologia , Proteínas da Matriz Extracelular/genética , Mutação , Fenótipo , Fator de Crescimento Transformador beta/genética , Idade de Início , Autoantígenos/genética , Lâmina Limitante Anterior/metabolismo , Distrofias Hereditárias da Córnea/genética , Epitélio Corneano/metabolismo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Colágenos não Fibrilares/genética , Linhagem , Recidiva , Microscopia com Lâmpada de Fenda , Colágeno Tipo XVIIRESUMO
Bietti's crystalline dystrophy (BCD) is an incurable retinal disorder caused by the polypeptide 2 of cytochrome P450 family 4 subfamily V (CYP4V2) mutations. Patients with BCD present degeneration of retinal pigmented epithelial (RPE) cells and consequent blindness. The lack of appropriate disease models and patients' RPE cells limits our understanding of the pathological mechanism of RPE degeneration. In this study, using CYP4V2 mutant pluripotent stem cells as disease models, we demonstrated that RPE cells with CYP4V2 mutations presented a disrupted fatty acid homeostasis, which were characterized with excessive accumulation of poly-unsaturated fatty acid (PUFA), including arachidonic acid (AA) and eicosapentaenoic acid (EPA). The PUFA overload increased mitochondrial reactive oxygen species, impaired mitochondrial respiratory functions, and triggered mitochondrial stress-activated p53-independent apoptosis in CYP4V2 mutant RPE cells. Restoration of the mutant CYP4V2 using adeno-associated virus 2 (AAV2) can effectively reduce PUFA deposition, alleviate mitochondria oxidative stresses, and rescue RPE cell death in BCD RPE cells. Taken together, our results highlight a role of PUFA-induced mitochondrial damage as a central node to potentiate RPE degeneration in BCD patients. AAV2-mediated gene therapy may represent a feasible strategy for the treatment of BCD.
Assuntos
Distrofias Hereditárias da Córnea/metabolismo , Células Epiteliais/metabolismo , Ácidos Graxos Insaturados/farmacologia , Mitocôndrias/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Células-Tronco Pluripotentes/metabolismo , Degeneração Retiniana/metabolismo , Doenças Retinianas/metabolismo , Epitélio Pigmentado da Retina/metabolismo , Animais , Células Cultivadas , Distrofias Hereditárias da Córnea/genética , Distrofias Hereditárias da Córnea/patologia , Família 4 do Citocromo P450/deficiência , Família 4 do Citocromo P450/genética , Células Epiteliais/patologia , Feminino , Técnicas de Inativação de Genes , Humanos , Camundongos , Camundongos SCID , Mitocôndrias/metabolismo , Mutação , Células-Tronco Pluripotentes/efeitos dos fármacos , Degeneração Retiniana/genética , Degeneração Retiniana/patologia , Doenças Retinianas/genética , Doenças Retinianas/patologia , Epitélio Pigmentado da Retina/patologiaAssuntos
Distrofias Hereditárias da Córnea/diagnóstico , Distrofias Hereditárias da Córnea/patologia , Cistos/diagnóstico , Cistos/patologia , Diagnóstico Diferencial , Oftalmopatias/diagnóstico , Oftalmopatias/patologia , Humanos , Limbo da Córnea/diagnóstico por imagem , Limbo da Córnea/patologia , Microscopia com Lâmpada de FendaRESUMO
ABSTRACT Lisch corneal dystrophy is a rare corneal disease characterized by the distinctive feature of highly vacuolated cells. Although this feature is important, the nature of these vacuoles within corneal cells remains unknown. Here, we sought to analyze corneal cells from a patient diagnosed with Lisch dystrophy to characterize the vacuoles within these cells. Analyses using histopathology examination, confocal microscopy, and transmission electron microscopy were all consistent with previous descriptions of Lisch cells. Importantly, the vacuoles within these cells appeared to be autophagosomes and autolysosomes, and could be stained with an anti-microtubule-associated protein 1A/1B-light chain 3 (LC3) antibody. Taken together, these findings indicate that the vacuoles we observed within superficial corneal cells of a patient with Lisch corneal dystrophy constituted autophagosomes and autolysosomes; this finding has not been previously reported and suggests a need for further analyses to define the role of autophagy in this ocular disease.
RESUMO A distrofia corneana de Lisch é uma doença rara, caracterizada principalmente pela presença de células altamente vacuoladas. Embora esta característica seja importante, a natureza desses vacúolos dentro das células da córnea permanece des conhecida. Aqui, procuramos analisar as células da córnea de um paciente diagnosticado com distrofia de Lisch para caracte rizar os vacúolos dentro dessas células. Análises utilizando exame histopatológico, microscopia confocal e microscopia eletrônica de transmissão foram todas consistentes com descrições previas de células de Lisch. Importante, os vacúolos dentro dessas células pareciam ser autofagossomos e autolisossomos, e po deriam ser corados com um anticorpo proteico 1A/1B-cadeia leve 3 (LC3) da proteína anti-microtúbulo associado a microtúbulos. Em conjunto, esses achados indicam que os vacúolos observados nas células superficiais da córnea de um paciente com distrofia corneana de Lisch constituíram autofagossomos e autolisossomos. Esse achado não foi relatado anteriormente e sugere a necessidade de mais análises para definir o papel da autofagia nessa doença ocular.
Assuntos
Humanos , Feminino , Adulto , Vacúolos/patologia , Distrofias Hereditárias da Córnea/patologia , Autofagossomos/patologia , Distrofias Hereditárias da Córnea/diagnóstico por imagem , Microscopia Confocal/métodos , Opacidade da Córnea/patologia , Opacidade da Córnea/diagnóstico por imagem , Tomografia de Coerência Óptica/métodos , Microscopia Eletrônica de Transmissão/métodos , MicroautofagiaRESUMO
Protein aggregation has been one of the leading triggers of various disease conditions, such as Alzheimer's, Parkinson's and other amyloidosis. TGFBI-associated corneal dystrophies are protein aggregation disorders in which the mutant TGFBIp aggregates and accumulates in the cornea, leading to a reduction in visual acuity and blindness in severe cases. Currently, the only therapy available is invasive and there is a known recurrence after surgery. In this study, we tested the inhibitory and amyloid dissociation properties of four osmolytes in an in-vitro TGFBI peptide aggregation model. The 23-amino acid long peptide (TGFBIp 611-633 with the mutation c.623 G>R) from the 4th FAS-1 domain of TGFBIp that rapidly forms amyloid fibrils was used in the study. Several biophysical methods like Thioflavin T (ThT) fluorescence, Circular Dichroism (CD), fluorescence microscopy and Transmission electron microscopy (TEM) were used to study the inhibitory and amyloid disaggregation properties of the four osmolytes (Betaine, Raffinose, Sarcosine, and Taurine). The osmolytes were effective in both inhibiting and disaggregating the amyloid fibrils derived from TGFBIp 611-633 c.623 G>R peptide. The osmolytes did not have an adverse toxic effect on cultured human corneal fibroblast cells and could potentially be a useful therapeutic strategy for patients with TGFBIp corneal dystrophies.
Assuntos
Amiloide , Córnea , Proteínas da Matriz Extracelular , Fibroblastos , Peptídeos , Agregação Patológica de Proteínas , Fator de Crescimento Transformador beta , Amiloide/química , Amiloide/genética , Amiloide/metabolismo , Linhagem Celular , Córnea/metabolismo , Córnea/patologia , Distrofias Hereditárias da Córnea/genética , Distrofias Hereditárias da Córnea/metabolismo , Distrofias Hereditárias da Córnea/patologia , Proteínas da Matriz Extracelular/química , Proteínas da Matriz Extracelular/genética , Proteínas da Matriz Extracelular/metabolismo , Fibroblastos/metabolismo , Fibroblastos/patologia , Humanos , Peptídeos/química , Peptídeos/genética , Peptídeos/metabolismo , Agregação Patológica de Proteínas/genética , Agregação Patológica de Proteínas/metabolismo , Agregação Patológica de Proteínas/patologia , Fator de Crescimento Transformador beta/química , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/metabolismoRESUMO
Purpose: To analyse the changing trends in penetrating keratoplasty indications between January 2011 and December 2018, at the Department of Ophthalmology, Saarland University Medical Center, Homburg/Saar, Germany. Patients and Methods: This is a retrospective review of 2123 corneal buttons of 1776 patients (1993 eyes, 56.0% males, age at the time of surgery 57.6 ± 18.7 years), who underwent penetrating keratoplasty (PKP) between January 2011 and December 2018. The classification was performed based on histological analysis and using the priority scheme of Brady et al., supplemented by a group of "failed endothelial keratoplasty grafts". Our groups were the following: pseudophakic or aphakic bullous keratopathy, regraft, failed endothelial keratoplasty graft, acute necrotizing and ulcerative keratitis, keratoconus, Fuchs' dystrophy, corneal dystrophy other than Fuchs', corneal scars and other diagnoses. Results: Between 2011 and 2018, keratoconus was the leading indication for PKP in 455 (21.5%) cases, followed by acute necrotizing and ulcerative keratitis in 384 (18.1%), regraft in 367 (17.3%), corneal scars in 350 (16.5%), pseudophakic or aphakic bullous keratopathy in 225 (10.6%), Fuchs' dystrophy in 194 (9.1%), other diagnoses in 64 (3.0%), corneal dystrophy other than Fuchs' in 52 (2.4%), and failed endothelial keratoplasty graft in 32 (1.5%) cases. Conclusions: With the introduction of posterior lamellar keratoplasty, keratoconus remains the leading PKP indication in our center with excimer laser-trephination on a routine basis. A trend towards increasing numbers can be observed regarding acute necrotizing and ulcerative keratitis patients and regrafts. However, the incidence of Fuchs' dystrophy decreased dramatically within PKP patients, with the introduction of posterior lamellar keratoplasty.
Assuntos
Doenças da Córnea/epidemiologia , Ceratoplastia Penetrante/tendências , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Doenças da Córnea/patologia , Doenças da Córnea/cirurgia , Distrofias Hereditárias da Córnea/epidemiologia , Distrofias Hereditárias da Córnea/patologia , Distrofias Hereditárias da Córnea/cirurgia , Úlcera da Córnea/epidemiologia , Úlcera da Córnea/patologia , Úlcera da Córnea/cirurgia , Feminino , Alemanha/epidemiologia , Sobrevivência de Enxerto , Humanos , Incidência , Lactente , Ceratocone/epidemiologia , Ceratocone/patologia , Ceratocone/cirurgia , Masculino , Pessoa de Meia-Idade , Reoperação , Estudos RetrospectivosRESUMO
Mutations in transforming growth factor-beta-induced (TGFBI) gene cause clinically distinct types of corneal dystrophies. To delineate the mechanisms driving these dystrophies, we focused on the R124C mutation in TGFBI that causes lattice corneal dystrophy type1 (LCD1) and generated novel transgenic mice harbouring a single amino acid substitution of arginine 124 with cysteine in TGFBI via ssODN-mediated base-pair substitution using CRISPR/Cas9 technology. Eighty percent of homozygous and 9.1% of heterozygous TGFBI-R124C mice developed a corneal opacity at 40 weeks of age. Hematoxylin and eosin and Masson trichrome staining showed eosinophilic deposits in subepithelial corneal stroma that stained negative for Congo-red. Although amyloid deposition was not observed in TGFBI-R124C mice, irregular amorphous deposits were clearly observed via transmission electron microscopy near the basement membrane. Interestingly, we found that the corneal deposition of TGFBI protein (TGFBIp) was significantly increased in homozygous TGFBI-R124C mice, suggesting a pathogenic role for the mutant protein accumulation. Furthermore, as observed in the LCD1 patients, corneal epithelial wound healing was significantly delayed in TGFBI-R124C mice. In conclusion, our novel mouse model of TGFBI-R124C corneal dystrophy reproduces features of the human disease. This mouse model will help delineate the pathogenic mechanisms of human corneal dystrophy.
Assuntos
Distrofias Hereditárias da Córnea/genética , Modelos Animais de Doenças , Proteínas da Matriz Extracelular/genética , Fator de Crescimento Transformador beta/genética , Substituição de Aminoácidos , Animais , Arginina/genética , Sistemas CRISPR-Cas , Distrofias Hereditárias da Córnea/patologia , Substância Própria/patologia , Substância Própria/ultraestrutura , Cisteína/genética , Feminino , Heterozigoto , Homozigoto , Humanos , Masculino , Camundongos , Camundongos Transgênicos , Microscopia Eletrônica de Transmissão , Mutação , Reparo de DNA por RecombinaçãoRESUMO
BACKGROUND: Hereditary gelsolin (AGel) amyloidosis is an autosomal dominantly inherited systemic amyloidosis that manifests with the characteristic triad of progressive ophthalmological, neurological and dermatological signs and symptoms. The National Finnish Gelsolin Amyloidosis Registry (FIN-GAR) was founded in 2013 to collect clinical data on patients with AGel amyloidosis, including altogether approximately one third of the Finnish patients. We aim to deepen knowledge on the disease burden and life span of the patients using data from the updated FIN-GAR registry. We sent an updated questionnaire concerning the symptoms and signs, symptomatic treatments and subjective perception on disease progression to 240 members of the Finnish Amyloidosis Association (SAMY). We analyzed the lifespan of 478 patients using the relative survival (RS) framework. RESULTS: The updated FIN-GAR registry includes 261 patients. Symptoms and signs corresponding to the classical triad of ophthalmological (dry eyes in 93%; corneal lattice amyloidosis in 89%), neurological (numbness, tingling and other paresthesias in 75%; facial paresis in 67%), and dermatological (drooping eyelids in 86%; cutis laxa in 84%) manifestations were highly prevalent. Cardiac arrhythmias were reported by 15% of the patients and 5% had a cardiac pacemaker installed. Proteinuria was reported by 13% and renal failure by 5% of the patients. A total of 65% of the patients had undergone a skin or soft tissue surgery, 26% carpal tunnel surgery and 24% at least unilateral cataract surgery. As regards life span, relative survival estimates exceeded 1 for males and females until the age group of 70-74 years, for which it was 0.96. CONCLUSIONS: AGel amyloidosis causes a wide variety of ophthalmological, neurological, cutaneous, and oral symptoms that together with repeated surgeries cause a clinically significant disease burden. Severe renal and cardiac manifestations are rare as compared to other systemic amyloidoses, explaining in part the finding that AGel amyloidosis does not shorten the life span of the patients at least for the first 75 years.
Assuntos
Neuropatias Amiloides Familiares/mortalidade , Neuropatias Amiloides Familiares/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Amiloidose/mortalidade , Amiloidose/patologia , Amiloidose Familiar/mortalidade , Amiloidose Familiar/patologia , Criança , Distrofias Hereditárias da Córnea/mortalidade , Distrofias Hereditárias da Córnea/patologia , Efeitos Psicossociais da Doença , Progressão da Doença , Feminino , Finlândia , Gelsolina , Humanos , Masculino , Pessoa de Meia-Idade , Sistema de Registros , Adulto JovemRESUMO
Collagen corneal cross-linking (CXL) is an invasive pharmacological treatment strategy used for corneal ectatic disorders and is currently the only treatment capable of halting the progression of the disease. In the past 20 years, the conservative management of progressive corneal ectasia has changed, thanks to this procedure that produces strengthening of the corneal tissue through the photochemical reaction generated by the combined action of riboflavin and ultraviolet A radiation. Many modified protocols have been implemented to halt the progression of the disease and to delay or prevent visual loss and surgical procedures such as corneal transplantation. Because of the variety of different protocols that are currently used, the results that are being reported are very variable, and could generate some confusion in relation to the true efficacy of the procedure. The aim of this review was to provide an overview of the aforementioned protocols that are designed to maintain the efficacy of CXL in halting the progression of the disease but avoiding the major limitations of the procedure.
Assuntos
Colágeno/uso terapêutico , Córnea/efeitos dos fármacos , Distrofias Hereditárias da Córnea/tratamento farmacológico , Reagentes de Ligações Cruzadas/uso terapêutico , Animais , Córnea/patologia , Distrofias Hereditárias da Córnea/patologia , HumanosRESUMO
Patients with gelatinous drop-like corneal dystrophy need to be effectively managed as the disease is severely debilitating in view of associated pho-tophobia and glare. Here, we report a rare case of gelatinous drop-like corneal dystrophy effectively managed by intraoperative anterior segment optical coherence tomography-guided manual deep anterior lamellar keratoplasty in 1 eye and sutureless fibrin glue-aided, microkeratome-assisted automated lamellar therapeutic keratoplasty in the other eye. The patient, a 22-year old man, presented with gradual diminution of vision associated with foreign body sensation, glare, photophobia, and watering due to corneal lesions, which were consistent with a diagnosis of gelatinous drop-like corneal dystrophy. Visual acuity at pre-sentation was 4/60 and 3/60 in the right and left eye, respectively. The patient received customized component lamellar keratoplasty in both eyes, and host tissue was sent for histopathologic examination. Treatment resulted in a best-corrected distance visual acuity of 6/9 and 6/12 in the right and left eye, respectively. The graft was clear and well apposed, with minimal interface haze bilaterally. The histopathologic report suggested intralamellar amyloid deposition in the form of homogenous, acellular eosinophilic deposits in the epithelium and anterior corneal stroma. This is a first report of the exclusive use of a fibrin-aprotinin tissue adhesive to stabilize a donor corneal lamellar graft as a treatment modality for a patient with gelatinous drop-like corneal dystrophy, suggesting that this treatment could supplant the need for sutures.