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Science Translational Medicine Jul 2019The Joslin Medalist Study characterized people affected with type 1 diabetes for 50 years or longer. More than 35% of these individuals exhibit no to mild diabetic...
The Joslin Medalist Study characterized people affected with type 1 diabetes for 50 years or longer. More than 35% of these individuals exhibit no to mild diabetic retinopathy (DR), independent of glycemic control, suggesting the presence of endogenous protective factors against DR in a subpopulation of patients. Proteomic analysis of retina and vitreous identified retinol binding protein 3 (RBP3), a retinol transport protein secreted mainly by the photoreceptors, as elevated in Medalist patients protected from advanced DR. Mass spectrometry and protein expression analysis identified an inverse association between vitreous RBP3 concentration and DR severity. Intravitreal injection and photoreceptor-specific overexpression of RBP3 in rodents inhibited the detrimental effects of vascular endothelial growth factor (VEGF). Mechanistically, our results showed that recombinant RBP3 exerted the therapeutic effects by binding and inhibiting VEGF receptor tyrosine phosphorylation. In addition, by binding to glucose transporter 1 (GLUT1) and decreasing glucose uptake, RBP3 blocked the detrimental effects of hyperglycemia in inducing inflammatory cytokines in retinal endothelial and Müller cells. Elevated expression of photoreceptor-secreted RBP3 may have a role in protection against the progression of DR due to hyperglycemia by inhibiting glucose uptake via GLUT1 and decreasing the expression of inflammatory cytokines and VEGF.
Topics: 3-O-Methylglucose; Acids; Animals; Cell Movement; Deoxyglucose; Diabetes Mellitus; Diabetic Retinopathy; Endothelial Cells; Ependymoglial Cells; Eye Proteins; Glycolysis; Humans; Intravitreal Injections; Mice, Inbred C57BL; Mice, Transgenic; Photoreceptor Cells, Vertebrate; Protective Agents; Protein Domains; Rats, Inbred Lew; Recombinant Proteins; Reproducibility of Results; Retina; Retinol-Binding Proteins; Signal Transduction; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factor Receptor-2; Vitreous Body
PubMed: 31270273
DOI: 10.1126/scitranslmed.aau6627 -
Investigative Ophthalmology & Visual... Jan 2023Pathologic myopia (PM) is one of the primary causes of blindness. This study aims to explore the possible relations between the composition of microRNA in vitreous...
PURPOSE
Pathologic myopia (PM) is one of the primary causes of blindness. This study aims to explore the possible relations between the composition of microRNA in vitreous exosomes of patients with PM and the progression of myopic maculopathy.
METHODS
Vitreous humor (VH) samples were collected from patients undergoing retinal surgery. A total of 15 and 12 VH samples were obtained from patients with PM and control, respectively. The PM group was divided into PM-L (G2) and PM-H groups (G3 and G4) in order to explore differentially expressed microRNAs (DEMs) that account for the relatively poor prognosis in G3 and G4 myopic maculopathy. A Weighted Gene Co-Expression Network Analysis (WGCNA) was conducted to find the persistently altered key microRNAs in myopic maculopathy progression. The Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genome (KEGG) pathway enrichment analysis were used.
RESULTS
High purity exosomes were extracted from the vitreous fluid of patients with PM and control. The top five downregulated DEMs of PM-H versus PM-L can reflect the tendency of deterioration of PM-H myopic maculopathy. MiR-143-3p and miR-145-5p, which were found in WGCNA, may participate in the development of myopic maculopathy. These microRNAs all relate to the insulin resistance pathway.
CONCLUSIONS
This is the first study to explore the relations between the progression of myopic maculopathy and vitreous exosomal microRNAs. Vitreous exosomal miR-143-3p and miR-145-5p can be considered biomarkers for patients with PM, and the vitreous exosomal DEM associated with PM-H may represent alarming signals of myopic maculopathy deterioration.
Topics: Humans; MicroRNAs; Myopia, Degenerative; Vitreous Body; Body Fluids; Retinal Diseases; Macular Degeneration; Exosomes
PubMed: 36648415
DOI: 10.1167/iovs.64.1.9 -
International Journal of Medical... May 2021We investigated whether nanopore amplicon sequencing of aqueous humor was capable of rapid pathogen identification in infectious endophthalmitis.
OBJECTIVES
We investigated whether nanopore amplicon sequencing of aqueous humor was capable of rapid pathogen identification in infectious endophthalmitis.
METHODS
5 cases of culture-positive bacterial endophthalmitis and 3 cases of fungal endophthalmitis (1 culture-positive and 2 presumed) were included. DNA was extracted from the aqueous humor and vitreous specimen, and PCR of bacterial rDNA (16S) and fungal rDNA (ITS1 and D1/2/3) was performed. Then, nanopore amplicon sequencing was performed for 2 h. The results of amplicon sequencing were compared to those of conventional culture studies.
RESULTS
In all cases, pathogens were identified by amplicon sequencing of aqueous humor specimens. In 3 cases of bacterial endophthalmitis, the identified microbes were confirmed by culture studies of both aqueous humor and vitreous specimens. In 2 cases of bacterial and 1 case of fungal endophthalmitis, the identified pathogens were confirmed only by culture studies of vitreous specimens. In all cases, amplicon sequencing identified pathogen in a shorter turnaround time than culture studies. In 2 cases with negative culture results, amplicon sequencing of aqueous humor identified fungal pathogens.
CONCLUSIONS
Our data demonstrates the potential of amplicon nanopore sequencing using aqueous humor to enable rapid, sensitive and less invasive microbial diagnosis of endophthalmitis.
Topics: DNA, Bacterial; Endophthalmitis; Humans; Nanopore Sequencing; Nanopores; Vitreous Body
PubMed: 33930723
DOI: 10.1016/j.ijmm.2021.151505 -
International Journal of Molecular... Feb 2021Photoreceptors are the light-sensing cells of the retina and the major cell type affected in most inherited retinal degenerations. Different metabolic pathways sustain...
Photoreceptors are the light-sensing cells of the retina and the major cell type affected in most inherited retinal degenerations. Different metabolic pathways sustain their high energetic demand in physiological conditions, particularly aerobic glycolysis. The principal metabolome of the mature retina has been studied, but only limited information is available on metabolic adaptations in response to key developmental events, such as eye opening. Moreover, dynamic metabolic changes due to retinal degeneration are not well understood. Here, we aimed to explore and map the ocular metabolic dynamics induced by eye opening in healthy (wild type) or -mutant (retinal degeneration 1, Rd1) mice, in which photoreceptors degenerate shortly after eye opening. To unravel metabolic differences emerging before and after eye opening under physiological and pathophysiological conditions, we performed nuclear magnetic resonance (NMR) spectroscopy-based metabolome analysis of wild type and Rd1 retina and vitreous/lens. We show that eye opening is accompanied by changes in the concentration of selected metabolites in the retina and by alterations in the vitreous/lens composition only in the retinal degeneration context. As such, we identify NAcetylaspartate as a potential novel vitreous/lens marker reflecting progressive retinal degeneration. Thus, our data can help elucidating mechanisms underlying key events in retinal physiology and reveal changes occurring in pathology, while highlighting the importance of the vitreous/lens in the characterization of retinal diseases.
Topics: Animals; Cyclic Nucleotide Phosphodiesterases, Type 6; Disease Models, Animal; Lens, Crystalline; Metabolome; Mice; Mutation; Retina; Retinal Degeneration; Vitreous Body
PubMed: 33652907
DOI: 10.3390/ijms22052345 -
Retina (Philadelphia, Pa.) Feb 2022The causes of floaters include posterior vitreous detachment and fundus hemorrhage, both of which are risk factors for retinal tears. We observed the vitreous of...
PURPOSE
The causes of floaters include posterior vitreous detachment and fundus hemorrhage, both of which are risk factors for retinal tears. We observed the vitreous of patients with floaters using swept source optical coherence tomography.
METHODS
Fundus examination was performed, and the vitreous was observed using swept source optical coherence tomography in 202 eyes of 202 patients with floaters. Patients with uveitis, diabetic retinopathy, and other fundus diseases were excluded.
RESULTS
Swept source optical coherence tomography revealed posterior vitreous detachment in 145 of 202 eyes (71.8%) and dot reflex like stardust in the vitreous in 42 of 202 eyes (20.8%). Posterior vitreous detachment occurred in 35 of 42 eyes (83.3%) and 110 of 160 eyes (68.8%) in the stardust (+) and stardust (-) groups, respectively; a significant difference was observed (P <0.001). In the stardust (+) group, 11 of 42 eyes (26.2%) had retinal tears with posterior vitreous detachment and 21 of 42 eyes (50.0%) had fundus hemorrhage. Three of 160 eyes (1.9%) and 4 of 160 eyes (2.5%) in the stardust (-) group had retinal tears with posterior vitreous detachment and fundus hemorrhage, respectively. Both tears and fundus hemorrhage were more frequent in the stardust (+) group than in the stardust (-) group (P <0.001).
CONCLUSION
The stardust sign on swept source optical coherence tomography indicates the risk of retinal tear.
Topics: Adult; Aged; Aged, 80 and over; Female; Fundus Oculi; Humans; Male; Middle Aged; Prospective Studies; Retinal Perforations; Tomography, Optical Coherence; Vitreous Body; Vitreous Detachment; Young Adult
PubMed: 35050930
DOI: 10.1097/IAE.0000000000003317 -
Asia-Pacific Journal of Ophthalmology... 2020
Topics: Anti-Bacterial Agents; Antifungal Agents; Bacteria; Coloring Agents; Endophthalmitis; Eye Infections, Bacterial; Eye Infections, Fungal; Humans; Microbial Sensitivity Tests; Staining and Labeling; Vitreous Body
PubMed: 31990737
DOI: 10.1097/01.APO.0000617904.11979.ae -
Ophthalmology. Retina Jan 2024Vitreoretinal lymphoma is a malignancy with high mortality. Incidence is rare, and there is a lack of medical evidence to direct management. This work describes... (Observational Study)
Observational Study
PURPOSE
Vitreoretinal lymphoma is a malignancy with high mortality. Incidence is rare, and there is a lack of medical evidence to direct management. This work describes presentation, diagnostic testing, and first treatment approaches in a recently diagnosed and treated patient cohort.
DESIGN
Clinical registry-based observational study.
SUBJECTS
Forty-eight women and 32 men (age range, 32-91 years; median age, 64 years) diagnosed with vitreoretinal lymphoma.
METHODS
An international network of ophthalmologists reported clinical features and management of patients presenting with vitreoretinal lymphoma between January 1, 2020 and December 31, 2022 via an electronic platform.
MAIN OUTCOME MEASURES
Visual acuity at presentation (logarithm of the minimum angle of resolution [logMAR]); basis for diagnosis; first treatment.
RESULTS
Vitreoretinal lymphoma was bilateral at presentation in 65% of patients (n = 52) and an initial site of lymphoma in 78% (n = 62). Of 127 eyes with lymphoma at presentation, vitreous was involved in 89% (n = 113) and was the only involved eye tissue in 40% (n = 51), and retina was involved in 46% (n = 59) and was the only involved eye tissue in 9% (n = 11). Median logMAR visual acuity of the worse-seeing eye was 0.50. The lymphoma was diagnosed from ocular specimens in 80% of patients (64/80), usually vitreous (57/64 patients [89%]), and on other clinical information in 20% of patients (16/80). Cellular studies were performed on ocular specimens from 59 of 64 patients (92%), most often cytology. Tumor gene analysis was used in 21 of 64 patients (33%), and cytokine assays were used in 13 of 64 patients (20%). For 76 patients (95%), treatment was initiated within 6 months of diagnosis and included ocular (38/76 [48%]), extraocular (17/76 [21%]), and ocular plus extraocular (21/76 [26%]) approaches. Intravitreal methotrexate was the most common ocular treatment (83/87 eyes [95%]).
CONCLUSIONS
Using data collected from 80 patients diagnosed with vitreoretinal lymphoma since 2020, we show that visual impairment is common, and that management often involves diagnosis by cellular tests and treatment with intravitreal chemotherapy.
FINANCIAL DISCLOSURE(S)
Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.
Topics: Male; Humans; Female; Middle Aged; Adult; Aged; Aged, 80 and over; Retinal Neoplasms; Vitreous Body; Eye Neoplasms; Lymphoma; Diagnostic Techniques and Procedures
PubMed: 37648063
DOI: 10.1016/j.oret.2023.08.012 -
Alzheimer's Research & Therapy Sep 2020Neurofilament light chain (NfL) is a promising biomarker of neurodegeneration in the cerebrospinal fluid and blood. This study investigated the presence of NfL in the...
BACKGROUND
Neurofilament light chain (NfL) is a promising biomarker of neurodegeneration in the cerebrospinal fluid and blood. This study investigated the presence of NfL in the vitreous humor and its associations with amyloid beta, tau, inflammatory cytokines and vascular proteins, apolipoprotein E (APOE) genotypes, Mini-Mental State Examination (MMSE) scores, systemic disease, and ophthalmic diseases.
METHODS
This is a single-site, prospective, cross-sectional cohort study. Undiluted vitreous fluid (0.5-1.0 mL) was aspirated during vitrectomy, and whole blood was drawn for APOE genotyping. NfL, amyloid beta (Aβ), total Tau (t-Tau), phosphorylated Tau (p-Tau181), inflammatory cytokines, chemokines, and vascular proteins in the vitreous were quantitatively measured by immunoassay. The main outcome measures were the detection of NfL levels in the vitreous humor and its associations with the aforementioned proteins. Linear regression was used to test the associations of NfL with other proteins, APOE genotypes, MMSE scores, and ophthalmic and systemic diseases after adjustment for age, sex, education level, and other eye diseases.
RESULTS
NfL was detected in all 77 vitreous samples. NfL was not found to be associated with ophthalmic conditions, APOE genotypes, MMSE scores, or systemic disease (p > 0.05). NfL levels were positively associated with increased vitreous levels of Aβ (p = 7.7 × 10), Aβ (p = 2.8 × 10), and t-tau (p = 5.5 × 10), but not with p-tau181 (p = 0.53). NfL also had significant associations with inflammatory cytokines such as interleukin-15 (IL-15, p = 5.3 × 10), IL-16 (p = 2.2 × 10), monocyte chemoattractant protein-1 (MCP1, p = 4.1 × 10), and vascular proteins such as vascular endothelial growth factor receptor-1 (VEGFR1, p = 2.9 × 10), Vegf-C (p = 8.6 × 10), vascular cell adhesion molecule-1 (VCAM-1, p = 5.0 × 10), Tie-2 (p = 6.3 × 10), and intracellular adhesion molecular-1 (ICAM-1, p = 1.6 × 10).
CONCLUSION
NfL is detectable in the vitreous humor of the eye and significantly associated with amyloid beta, t-tau, and select inflammatory and vascular proteins in the vitreous. Additionally, NfL was not associated with patients' clinical eye condition. Our results serve as a foundation for further investigation of NfL in the ocular fluids to inform us about the potential utility of its presence in the eye.
Topics: Alzheimer Disease; Amyloid beta-Peptides; Biomarkers; Cross-Sectional Studies; Humans; Intermediate Filaments; Neurofilament Proteins; Prospective Studies; Vascular Endothelial Growth Factor A; Vitreous Body; tau Proteins
PubMed: 32943089
DOI: 10.1186/s13195-020-00677-4 -
Journal of Acquired Immune Deficiency... May 2023To determine tenofovir (TFV) penetration into intraocular tissues using ultra high-performance liquid chromatography/tandem mass spectrometry (UHPLC-MS/MS). (Observational Study)
Observational Study
OBJECTIVE
To determine tenofovir (TFV) penetration into intraocular tissues using ultra high-performance liquid chromatography/tandem mass spectrometry (UHPLC-MS/MS).
METHODS
Nineteen participants taking tenofovir in combination antiretroviral therapy (cART) regimen who underwent pars plana vitrectomy (PPV) surgery were enrolled in the observational retrospective study between January 2019 and August 2021. The participants were divided into mild, moderate, and severe groups according to retinal manifestations. Basic information was recorded during PPV surgery. Paired blood plasma and vitreous humor samples (n = 19) were collected for UHPLC-MS/MS.
RESULTS
The median plasma and vitreous tenofovir concentrations were 106.00 ng/mL (interquartile range[IQR], 54.6-142.5) and 41.40 ng/mL (IQR 9.4-91.6), respectively. The median vitreous/plasma concentration ratio from the paired samples was 0.42 (IQR 0.16-0.84). The plasma and vitreous tenofovir concentrations were significantly correlated (r = 0.483, P = 0.036). The median vitreous tenofovir concentration was the lowest in the mild group (4.58 ng/mL). Six vitreous samples were below 50% inhibitory concentration (IC50) (11.5 ng/mL), and 2 of them were undetectable. Significant differences were noted in vitreous/plasma and vitreous tenofovir concentrations ( P = 0.035 and P = 0.045, respectively) among the 3 groups but not in plasma tenofovir concentration ( P = 0.577). No correlation was noted between vitreous HIV-1 RNA and vitreous tenofovir concentrations (r = 0.049, P = 0.845).
CONCLUSION
Vitreous tenofovir did not reliably or consistently achieve concentrations sufficient to inhibit viral replication in intraocular tissues due to poor penetration of the blood-retinal barrier (BRB). The higher vitreous tenofovir concentrations were associated with moderate or severe disease compared with mild disease, indicating an association with the severity of BRB disruption.
Topics: Humans; Anti-HIV Agents; HIV Infections; Retrospective Studies; Tandem Mass Spectrometry; Tenofovir; Vitrectomy; Vitreous Body
PubMed: 36881850
DOI: 10.1097/QAI.0000000000003171 -
System-wide vitreous proteome dissection reveals impaired sheddase activity in diabetic retinopathy.Theranostics 2022Diabetic retinopathy (DR) is a major complication of diabetes mellitus causing significant vision loss. DR is a multifactorial disease involving changes in retinal...
Diabetic retinopathy (DR) is a major complication of diabetes mellitus causing significant vision loss. DR is a multifactorial disease involving changes in retinal microvasculature and neuronal layers, and aberrations in vascular endothelial growth factors (VEGF) and inflammatory pathways. Despite the success of anti-VEGF therapy, many DR patients do not respond well to the treatment, emphasizing the involvement of other molecular players in neuronal and vascular aberrations in DR. We employed advanced mass spectrometry-based proteome profiling to obtain a global snapshot of altered protein abundances in vitreous humor from patients with proliferative DR (PDR) in comparison to individuals with epiretinal membrane without active DR or other retinal vascular complications. Global proteome correlation map and protein-protein interaction networks were used to probe into the functional inclination of proteins and aberrated molecular networks in PDR vitreous. In addition, peptide-centric analysis of the proteome data was carried out to identify proteolytic processing, primarily ectodomain shedding events in PDR vitreous. Functional validation experiments were performed using preclinical models of ocular angiogenesis. The vitreous proteome landscape revealed distinct dysregulations in several metabolic, signaling, and immune networks in PDR. Systematic analysis of altered proteins uncovered specific impairment in ectodomain shedding of several transmembrane proteins playing critical roles in neurodegeneration and angiogenesis, pointing to defects in their regulating sheddases, particularly ADAM10, which emerged as the predominant sheddase. We confirmed that ADAM10 protease activity was reduced in animal models of ocular angiogenesis and established that activation of ADAM10 can suppress endothelial cell activation and angiogenesis. Furthermore, we identified the impaired ADAM10-AXL axis as a driver of retinal angiogenesis. We demonstrate restoration of aberrant ectodomain shedding as an effective strategy for treating PDR and propose ADAM10 as an attractive therapeutic target. In all, our study uncovered impaired ectodomain shedding as a prominent feature of PDR, opening new possibilities for advancement in the DR therapeutic space.
Topics: Animals; Diabetes Mellitus; Diabetic Retinopathy; Peptide Hydrolases; Proteome; Vascular Endothelial Growth Factors; Vitreous Body
PubMed: 36185601
DOI: 10.7150/thno.72947