Proven and less studied hematopoietic and vasoactive growth factors in retinal capillary hemangioma

Pathogenesis of retinal capillary hemangioma has not been sufficiently studied at the present time. Therefore, the study of cytokine levels in biological fluids seems to be very relevant in order to increase knowledge about the mechanisms of the disease development and searching for targeted therapies. The content of hematopoietic and vasoactive growth factors in blood serum, lacrimal fluid, and vitreous body was studied in patients with retinal capillary hemangioma. A total of 26 patients with retinal angiomatosis were examined. The samples of blood serum (n = 23) and lacrimal fluid (n = 10) from practically healthy people aged 22 to 46 (27.4±1.4 years) were used as a control. To perform comparative assessment of cytokine concentrations in the vitreous body of patients with retinal capillary hemangioma, were used samples of the vitreous body from 6 patients (average age 33±4.7 years; from 21 to 49 years) with rhegmatogenous retinal detachment. To measure the cytokine concentrations, we applied multiplex analysis technique using the xMAP platform with LuminexxPONENT 3.1 program and ProcartaPlex sets (eBioscience, Austria). A detailed characteristic of vasoactive factors in capillary retinal hemangioma was obtained as a result of this work. Some disorders in chemokine regulation were identified. There was a significant increase in serum concentrations of three vasoactive factors, i.e., PDGF-BB, HGF, and PIGF-1, with a decrease in chemokines (MCP-1, MIP-1α, and MIP-1β). The frequencies of PIGF-1 and MIP-1α detection also significantly differed from the control group. SCF was significantly more often determined in patients with retinal angiomatosis only at the systemic level. Correlations between PDGF-BB and PIGF-1, as well as PIGF-1 and MIP-1β were shown. A significant increase in VEGF-A, HGF, VEGF-D, as well as MCP-1 concentrations was shown in the lacrimal fluid. The inversion of PDGF-BB concentrations in serum and lacrimal fluid was noted. Analysis of intraocular cytokine levels revealed a significant increase in VEGF-A and HGF concentrations, with marked decrease in MIP-1α and MIP-1β. PDGF-BB in 100% of cases was determined only in vitreous body of patients with retinal angiomatosis. With respect to the revealed characteristic shifts of HGF/SF intraocular production in retinal capillary hemangioma, it seems relevant to search ways for its inhibition, thus providing potential basis for a new therapeutic strategy in treatment of retinal angiomatosis

Screening of cytokines in blood serum and lacrimal liquid in wet and atrophic forms of age-related macular degeneration

Cytokines play an integral role in pathogenesis of age-related macular degeneration (AMD). Of particular interest are the late stages of this disease, which causes progressive visual impairment. Therapy-induced effects of post-treatment cytokine concentrations also need to be studied, both at long and short observation terms. These studies are of vital importance if the atrophy occurs during antiangiogenic therapy. Our purpose was to study an array of 45 cytokines, in blood serum (BS) and lacrimal liquid (LL) of the patients with wet and atrophic AMD.The study included 70 people (85 eyes) with stage 3-4 AMD according to AREDS. Depending on the form of AMD, 3 groups were discerned: I group (n = 24) included the patients with “geographic atrophy”; II group (n = 22), consisted of the patients with macular atrophy treated with antiangiogenic therapy of wet AMD; III group (n = 24), comprised the patients with a wet AMD who did not previously receive the treatment. Control group consisted of healthy volunteers (n = 25). All the groups were comparable for age and gender. The patients underwent a comprehensive ophthalmological examination to make a diagnosis. A multiplex study of the local (in the BS) and systemic (in the LL) cytokine status was carried out on a MAGPIX device (platform хMAP, Luminex Corporation, USA) in the Luminexx PONENT 3.1 software, using Procarta Plex kits (eBioscience, Austria). We determined 45 cytokines causing various biological effects, i.e., IL-1á, IL-1â, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12p70, IL-13, IL-15, IL-17A, IL-18, IL-21, IL-22, IL-23, IL-27, IL-31, IFNá, IFNã, IL-8/CXCL8, IP-10/CXCL10, SDF-1á/CXCL12, MCP-1/CCL2, MIP-1á/CCL3, MIP-1â/ CCL4, RANTES/CCL5, Eotaxin/CCL11, TNFá, TNFâ, GM-CSF, VEGF-A, VEGF-D, FGF-2, EGF, PDGF-BB, HGF, SCF, GRO-á, NGF-â, BDNF, LIF, PIGF-1.Screening of a wide range of cytokines showing various biological effects was carried out in BS and LL of patients with atrophic and wet forms of AMD. It has been shown that the late stages of the disease are associated with local and systemic changes of pro / anti-inflammatory mediators (IL-1â, IL-1ra, IL-18, LIF), chemoattractant cytokines (IL-8/CXCL8, IP-10/CXCL10, MCP-1/CCL2, MIP-1á/CCL3, MIP-1â/ CCL4, RANTES/CCL5, Eotaxin/CCL11), hematopoietic regulators (IL-7), and growth factors with known angiogenic activity (EGF, HGF, PDGF-BB, VEGF-A). Altered concentrations of numerous chemokines, e.g., IP-10/CXCL10, SDF-1á/CXCL12, MIP-1á/CCL3, MIP-1â/CCL4, RANTES/CCL5 and Eotaxin/CCL11 (p < 0.05) in BS of the patients with atrophic and wet AMD may be of interest for the search of biomarkers associated with various clinical phenotypes of the disease and may be also helpful for development of new therapeutic strategies

EXAMINING LOCALLY EXPRESSED mRNA OF INFLAMMATORY MEDIATOR GENES IN A MODEL OF RETINAL PIGMENT EPITHELIUM ATROPHY AND RETINAL DEGENERATION INDUCED BY SUBRETINAL SALINE INJECTION IN RABBITS

Degenerative-dystrophic retinal diseases, particularly age-related macular degeneration (AMD), are now considered to be the lead cause of blindness and low vision in developed countries, with a steadily increasing trend. Recent publications provide evidence for the involvement of inflammatory mechanisms in TMD development and progression unveiled due to advances in innate and adaptive immunity research. However, the immunopathogenesis of atrophic AMD form, “geographic atrophy” (GA) remains largely unstudied. Objective: to investigate local mRNA expression of inflammatory cytokines IL-1β, IL-18, CCL2/MCP-1 in a model of RPE atrophy induced after subretinal injection of 0.9% sodium chloride solution in experimental rabbits. The investigation was carried out in tissue complex retina-RPE-choroid (TC) samples isolated from eyes of 23 albino New Zealand rabbits after modeling RPE atrophy by subretinal injection of 0.9% sodium chloride solution and 5 healthy rabbits lacking eye lesions. Animals in the experimental group (one week before surgical intervention, in the early period, and in the period of sustained RPE atrophy formation) and controls were subjected to optical coherence tomography (OCT) and ocular fundus autofluorescence (FAF). Evaluation of proinflammatory cytokine gene expression levels in TC was performed by RT-PCR. Results. Subretinal injection of 0.01 ml of 0.9% sodium chloride solution induced experimental RPE atrophy development in rabbits vs. control that was associated with multidirectional changes of IL-1β, IL-18, MCP-1/CCL2 gene mRNA expression. Three types of response in the TC, formed during development of atrophic changes and determined by the value of local cytokine gene expression were characterized: 1) hypo/ no response – decreased/no expression; 2) normal response – moderate increase; 3) hyper response – overexpression. 69.6% of animals with persistent atrophy had a moderate to hypertrophic increase in locally expressed mRNA MCP-1/CCL2, whereas 30% cases had significantly increased IL-1β mRNA expression – factors damaging the blood-retinal barrier and contributing to posterior segment immune privilege. It should be taken into account while developing new strategies for treatment of ophthalmic pathology, in particular the currently actively studied and tested options for RPE stem cell transplantation into subretinal space. The data obtained may be useful to investigate various types of RPE atrophy and develop new strategies of ophthalmopathology treatment in preclinical studies

Surgical treatment of von Hippel-Lindau disease complicated by retinal detachment: a clinical case

A clinical case of surgical treatment of a patient with von Hippel-Lindau disease complicated by retinal detachment is presented. Intravitreal injection of ranibizumab was performed 7 days before vitrectomy. Laser photodestruction of retinal capillary hemangiomas was performed intraoperatively with an increased exposure and reduced inter-pulse interval. A 1 year follow-up showed a stable functional and anatomic result, and lack of relapse. Details of surgical treatment and advantages of the method proposed are discussed. For citation: Neroev V.V., Ilyukhin P.A., Ryabina M.V., Novikova A.Yu. Surgical treatment of von Hippel-Lindau disease complicated by retinal detachment: a clinical case. Russian ophthalmological journal. 2018; 11 (1):85-92. doi: 10.21516/2072-0076-2018-11-1-85-92 (In Russian)

Assessment of effect of intravitreal ranibizumab injection on the ocular blood flow in patients with neovascular age-related macular degeneration

Purpose: To evaluate the effect of intravitreal ranibizumab injection on the ocular blood flow in patients with neovascular agerelatedmacular degeneration (AMD).Methods: 35 patients with wet AMD undergone intravitreal 0.5 mg ranibizumab injection. Color Doppler Imaging (CDI) and dopplerographywere used to measure hemodynamic parameters including the peak systolic velocity (Vsyst, cm / s), end-diastolic velocity (V diast, cm / s), and resistance index (RI) of blood flow in the central retinal artery (CRA), the short posterior ciliary arteries (PCA), and the ophthalmic artery (OA). All patients were examined before and after injection on day 1‑7 and 30 day during the 3‑month follow up period.Results: Before intravitreal injection Vsyst was decreased in short PCA (p<0.05), RI in CRA and in short PCA significantly increased in comparison with normal index in same vessels. The peak systolic velocity in OA, in CRA and in short PCA was not significantly changed. After second injection resistance index in CRA and in short PCA was normalized.Conclusion: There was not impairment of ocular blood flow in retinal and choroidal after monthly intravitreal injection of ranibizumab during the 3‑month follow up period

CYTOKINES IN LACRIMAL FLUID AND BLOOD SERUM: EARLY BIOMARKERS OF AGE-RELATED MACULAR DEGENERATION

The article presents results of multiplex cytokine assays in blood serum and lacrimal fluid at the initial and intermediate stages of age-related macular degeneration (AMD). Some features of local and systemic disturbances in the cytokine profile were detected in these patients. It was revealed that the initial stage of AMD was associated with elevated IL-17 levels in lacrimal fluid, along with imbalance between the local increase and systemic decrease of TGF-β1 amounts. Intermediate-stage AMD was associated with increased levels of the most cytokines assayed (except of TGF-β1) in blood serum and lacrimal fluid, thus suggesting stimulation of both pro-inflammatory and angiogenic responses, like as activation of anti-inflammatory and anti-infective factors

Modeling the atrophy of retinal pigment epithelium

Purpose: to develop easy-to make and reproducible models of retinal pigment epithelium atrophy (RPE) and retinal degeneration using two types of solution (0.9 % sodium chloride and bevacizumab) and to evaluate these models using clinical instrumental and pathomorphological studies. Material and methods. To create the two models, we used 60 New Zealand albino rabbits divided into 2 groups of 30 animals each (30 eyes). In group 1, 0.01 ml of 0.9 % sodium chloride solution was delivered into the subretinal space at a distance of 1–1.5 mm downwards from the optic disc forming a subretinal bladder, whilst group 2 received 0.01 ml of bevacizumab solution which contained 0.025 mg of the drug. Optical coherence tomography (OCT) and fundus autofluorescence imaging were performed in live rabbits’ eyes before and after the procedure on the 2nd, 7th, 14th, 24th, and 30th day using Heidelberg Spectralis™ SD-OCT (Heidelberg Engineering, Germany). The enucleated eyes were histologically evaluated 14 and 30 days after RPE atrophy modeling. Results. Two easily reproducible experimental models of RPE atrophy have been developed. Clinical and morphological indications of RPE atrophy are described. Histological analysis revealed a more aggressive action of 0.9% sodium chloride solution on the retina and the choroid as compared with the model obtained with a similarly delivered subretinal angiogenesis inhibitor. Conclusion. The obtained experimental models may be useful in investigating various types of RPE atrophy, including those arising from the use of angiogenesis inhibitors

Differential diagnosis of retinal capillary hemangiomas and vasoproliferative tumors

Purpose. To propose the major differential diagnostic criteria of retinal capillary hemangioma (RCH) and vasoproliferative tumor (VPT) aimed at increasing the efficiency of the diagnosis of these diseases.Material and methods. A comprehensive ophthalmologic examination off 19 (26 eyes) patients, aged 19 to 66 (ave. 31.89 ± 11.30 yrs), with vascular neoplasms of peripheral localization included, beside the standard techniques, fundus photoregistration, fluorescent angiography (FAG), optical coherence tomography of the retina and a combined ultrasound testing using B scanning, echographic densitometry, colour Doppler imaging (CDI), and pulse Doppler imaging.Results. Based on the analysis of the clinical picture and the results of the tests performed, the following differential diagnostic criteria were proposed: the age at which RCH and VPT developed, their localization in the eye fundus, and the value of the maximal systolic blood flow velocity in the central vein of the retina.Conclusion. Differential diagnostics of RCH and VPT requires combined approaches and should take into account, in addition to case history and the clinical picture, also the results of instrumental testing techniques: FAG, echography and CDI

Association of mutations and polymorphic variants of the VHL gene with the clinical characteristics of retinal capillary hemangioma

Purpose. To study the association of mutations and polymorphic variants of the VHL gene with the clinical characteristics of retinal capillary hemangioma.Material and methods. 29 DNA samples isolated from peripheral blood leukocytes of patients with retinal capillary hemangiomas were tested. The analysis of mutations and polymorphic markers of the VHL gene was carried out by high resolution melting curve analysis.Results. A significant correlation was found between the frequency of tyrosine mutation in the 156th position of the VHL gene and the localization of the capillary hemangioma in the middle periphery of the fundus, the presence of local exudative retinal detachment and fibroglial proliferation in the tumor region (p = 0.02; RR = 1.86). An association was established between arginine mutation frequency in the 200th position of the gene with the presence of cerebellar hemangioblastoma and spinal cord. A statistically significant association of the GA genotype of the polymorphic marker rs779805 of the VHL gene with the localization of capillary hemangioma in the middle periphery of the fundus was established (p = 0.03; RR = 4.8). It has been shown that the carriage of the T allele of the polymorphic marker rs779808 may be associated with the characteristic localization of the tumor at the extreme periphery of the retina in the upper temporal quadrant. A tendency to increasing frequency of occurrence of the AC genotype of the polymorphic marker rs1678607 in patients with hemorrhagic complications of capillary hemangioma, as well as to increasing frequency of the TT genotype of the polymorphic marker rs1642742 in patients with cerebellar hemangioblastomas was determined.Conclusion. Further research involving more clinical material is needed, as our results may be important for early diagnosis of Hippel — Lindau disease, prediction of the onset and clinical course of retinal capillary hemangiomas, as well as of the probability of systemic manifestations of the disease

Possibilities of Complex Ultrasound Examination in the Assessment of Hemodynamic and Structural Features of Retinal Capillary Hemangioma

Purpose: to study the possibilities of complex ultrasound examination in the assessment of hemodynamic and structural features of retinal capillary hemangioma (RCH).Patients and methods. 26 patients (35 eyes) with RCHs at the age of 11 to 53 years (mean 26.6 ± 9.4 years) were examined. Ultrasound examination was performed using B–mode image, echodensitometry, Color Doppler Imaging (CDI) and pulsed Doppler (PD). The peak systolic velocity (Vsyst), the end-diastolic velocity (Vdiast) of the blood flow, and the resistance index (RI) in the orbital vessels were examined using CDI and PD. Qualitative and quantitative assessment of blood flow in a feeding artery and a draining vein of the RCHs were performed. The follow-up efficiency evaluation of the laser coagulation of RCH was performed in 12 eyes. Results. The elevation of RCH was 1.7 ± 0.7 mm (from 0.5 to 3.3 mm) on average, the base diameter — 5.4 ± 2.1 mm (from 1.8 to 11.5 mm). The acoustic density varied from 42 to 176 (mean 109.6 ± 30.9) in relative units. The indices of Vsyst were from 5.5 to 21.4 cm/s (mean value — 10.6 ± 4.2 cm/s) in the feeding arteries, from 3.06 to 14.3 cm/s — in veins (mean value — 4.9 ± 2.4 cm/s). A significant increase in the blood flow velocity in the central retinal vein (CRV), a decrease in RI in the central retinal artery (CRA) and short posterior ciliary arteries (SPCAs) were determined. In 3 (8.6 %) eyes, in the absence of echographic manifestations of small RCHs of optic nerve head (ONH), there was an increase of the blood flow velocity in the CRV (the mean value was 8.95 ± 1.15 cm/s). After treatment, B-scan ultrasonogram showed no visualization of RCHs in 3 eyes. A significant decrease (31 %) of prominence compared to the baseline values was registered in 9 eyes (66.7 %). A decrease of the tumor base the diameter by an average of 37.7 % was also registered in 7 eyes (58.3 %). A significant decrease (26.3 %) in Vsyst in arterial-type feeding vessels was determined. Conclusion. Significant changes in regional ocular blood flow in RCH were registered. The increase of blood flow in the CRV at the localization of RCH on the ONH can be an additional differential diagnostic criterion of the disease. The hemodynamic instability in the retinal vessels feeding the RCH and the positive dynamics of the blood flow after laser coagulation were determined