Characterization of the Vitreous Proteome in Diabetes without Diabetic Retinopathy and Diabetes with Proliferative Diabetic Retinopathy

Gao, Benbo; Chen, Xiaohong; Timothy, Nigel H.; Aiello, Lloyd Paul; Feener, Edward P.

doi:10.1021/pr800112g

Cited by 215 publications

(251 citation statements)

References 34 publications

Supporting

Mentioning

234

Contrasting

Order By: Relevance

“…The vitreous levels of RAS components, including prorenin, angiotensinogen and angiotensin II, have been reported to increase in PDR eyes [33,34,40,41]. We confirmed the increased levels of prorenin (Fig.…”

Section: Discussionsupporting

confidence: 82%

See 1 more Smart Citation

(Pro)renin receptor is associated with angiogenic activity in proliferative diabetic retinopathy

et al. 2012

View full text Add to dashboard Cite

Aims/hypothesis The renin-angiotensin system (RAS) potentially has a role in the development of end-organ damage, and tissue RAS activation has been suggested as a risk factor for diabetic retinopathy. We have recently shown significant involvement of (pro)renin receptor ([P]RR) in retinal inflammation in a rodent model of early diabetes. In this study we aim to elucidate the (P)RR-associated pathogenesis of fibrovascular proliferation, a late-stage angiogenic complication in human diabetic retinopathy. Methods Vitreous fluids from 23 eyes of patients with proliferative diabetic retinopathy (PDR) and 16 eyes of controls with non-diabetic, idiopathic macular diseases (macular hole and epiretinal membrane) were collected. Protein levels of soluble (P)RR were measured by ELISA, and immunofluorescence was performed to assess the localisation of (P)RR and related molecules in fibrovascular tissues from PDR eyes. Results (P)RR immunoreactivity was detected in neovascular endothelial cells, colocalised with prorenin, phosphorylated extracellular signal-regulated kinase (ERK) and vascular endothelial growth factor (VEGF). Prorenin application to human retinal microvascular endothelial cells significantly upregulated mRNA expression of VEGF, especially the VEGF165 isoform, which was abolished by (P)RR or ERK signalling blockade. Proteases known to cleave (P)RR, including furin, were positive in endothelial cells in fibrovascular tissues. Protein levels of soluble (P)RR in vitreous fluids were higher in PDR eyes than in non-diabetic control eyes, and correlated significantly with vitreous prorenin and VEGF levels and the vascular density of fibrovascular tissues. Conclusions/interpretation Our data using human samples provide the first evidence that (P)RR is associated with angiogenic activity in PDR.

show abstract

Section: Discussionsupporting

confidence: 82%

“…Activated prorenin (Fig. 6c) together with angiotensinogen [34,41] is thought to trigger tissue RAS activation causing angiotensin II generation [34,41] in PDR. Therefore s(P)RR accumulation in vitreous fluids, in concert with fl(P)RR in fibrovascular tissues, is likely to enhance tissue RAS activation in PDR.…”

Section: Discussionmentioning

confidence: 99%

(Pro)renin receptor is associated with angiogenic activity in proliferative diabetic retinopathy

et al. 2012

View full text Add to dashboard Cite

show abstract

“…Some previous studies pointed to reductions in the levels of vitreous PEDF in patients suffering from PDR [6,18-20]. Conversely, elevated levels of PEDF were detected in some studies [13,21]. We have detected reduced levels of PEDF in the vitreous fluid of diabetic patients with PDR by both proteomic analysis and Western blotting.…”

Section: Discussionmentioning

confidence: 43%

“…Previous studies have identified some proteins that are differentially expressed in the vitreous of PDR patients. Many of the proteins are involved in such processes as angiogenesis, cellular proliferation, acute phase response, vascular permeability changes, and oxygen-induced vessel loss [7,13]. These studies involved the use of vitreous from non-diabetic patients with macular hole (MH) or macular epiretinal membrane (MEM) as control groups.…”

Section: Introductionmentioning

confidence: 99%

Characterisation of the vitreous proteome in proliferative diabetic retinopathy

Wang

Hu³

et al. 2012

Proteome Sci

View full text Add to dashboard Cite

BackgroundDiabetes can lead to serious microvascular complications such as proliferative diabetic retinopathy (PDR), which results in severe vision loss. The diabetes-induced alterations in the vitreous protein composition in diabetic patients with PDR may be responsible for the presence of PDR. The vitreous humour can be utilised in a variety of studies aimed toward the discovery of new targets for the treatment or prevention of PDR and the identification of novel disease mechanisms. The aim of this study was to compare the protein profile of vitreous humour from diabetic patients with PDR with that of vitreous humour from normal human eyes donated for corneal transplant.ResultsVitreous humour from type 2 diabetic patients with PDR (n = 10) and from normal human eyes donated for corneal transplant (n = 10) were studied. The comparative proteomic analysis was performed using two-dimensional fluorescence difference gel electrophoresis (2-D DIGE). Differentially produced proteins (abundance ratio > 2 or < -2, p < 0.01) were identified by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF MS) and MALDI-TOF tandem mass spectrometry. A total of 1242 protein spots were detected on the 2-D master gel of the samples, and 57 spots that exhibited statistically significant variations were successfully identified. The spots corresponded to peptide fragments of 29 proteins, including 8 proteins that increased and 21 proteins that decreased in PDR. Excluding the serum proteins from minor vitreous haemorrhage, 19 proteins were found to be differentially produced in PDR patients compared with normal subjects; 6 of these proteins have never been reported to be differentially expressed in PDR vitreous: N(G),N(G)-dimethylarginine dimethylaminohydrolase 1 (DDAH 1), tubulin alpha-1B chain, gamma-enolase, cytosolic acyl coenzyme A thioester hydrolase, malate dehydrogenase and phosphatidylethanolamine-binding protein 1 (PEBP 1). The differential production of pigment epithelium-derived factor (PEDF) and clusterin was confirmed by Western blot analysis.ConclusionsThese data provide an in-depth analysis of the human vitreous proteome and reveal protein alterations that are possibly involved in the pathogenesis of PDR. Further investigation of these special proteins may provide potential new targets for the treatment or the prevention of PDR.

show abstract

“…1,2 Protein biomarkers in the vitreous core have been associated with diseases such as diabetic retinopathy. [3][4][5] How these proteins are differentially expressed in each of the substructures, and in many cases the specific protein identities, are not known. These details may give insight to the origin of proteins associated with specific vitreoretinal diseases and help target future therapies.…”

Section: Discussionmentioning

confidence: 99%

Dissection of Human Vitreous Body Elements for Proteomic Analysis

Skeie¹,

Mahajan²

2011

JoVE

View full text Add to dashboard Cite

The vitreous is an optically clear, collagenous extracellular matrix that fills the inside of the eye and overlies the retina.1,2 Abnormal interactions between vitreous substructures and the retina underlie several vitreoretinal diseases, including retinal tear and detachment, macular pucker, macular hole, age-related macular degeneration, vitreomacular traction, proliferative vitreoretinopathy, proliferative diabetic retinopathy, and inherited vitreoretinopathies. 1,2 The molecular composition of the vitreous substructures is not known. Since the vitreous body is transparent with limited surgical access, it has been difficult to study its substructures at the molecular level. We developed a method to separate and preserve these tissues for proteomic and biochemical analysis. The dissection technique in this experimental video shows how to isolate vitreous base, anterior hyaloid, vitreous core, and vitreous cortex from postmortem human eyes. One-dimensional SDS-PAGE analyses of each vitreous component showed that our dissection technique resulted in four unique protein profiles corresponding to each substructure of the human vitreous body. Identification of differentially compartmentalized proteins will reveal candidate molecules underlying various vitreoretinal diseases. Video LinkThe video component of this article can be found at https://www.jove.com/video/2455/ Protocol 1. Anterior Segment Dissection.1. The cornea is removed by first making an incision at the limbus into the anterior chamber using a 15° blade (Figure 1). Then one blade of curved cornea-scleral scissors is inserted into the anterior chamber. Circumferential cuts are made in the cornea, just anterior to the limbus. 2. 0.12 Colibri forceps and Westcott scissors are used to cut the iris circumferentially anterior to the ciliary body. 3. The lens nucleus is removed with 0.12 Colibri forceps or a 15° blade (supersharp) and the cortex and capsule with forceps. Vitreous Core Aspiration.1. A 23-gauge needle on a 5-cc syringe is inserted into the mid-vitreous (Figure 1). 2. Approximately 1 mL or more of vitreous core is gently aspirated. 3. The sample is then placed in a microfuge tube and then into liquid nitrogen. Anterior Hyaloid Dissection.1. The anterior hyaloid is seen as a semi-transparent ring by adjusting the incident light from a goose neck microscope light. Previous aspiration of the vitreous core separates the anterior hyaloid from other structures for easier identification. 2. The sheet of elastic tissue is carefully pulled away from the ciliary body with Colibri or dressing forceps and cut with Vannas scissors. This maneuver is repeated. 3. The sample is then placed in a microfuge tube and then into liquid nitrogen. Vitreous Base Dissection.1. Using 0.12 forceps to stabilize the eye cup, Westcott scissors are used to "flower" the eye by making four equally spaced stress-relieving cuts from the ciliary body towards the optic nerve head. 2. The pars plicata of the ciliary body (Figure 2) is removed from each of the quadrants using ...

show abstract

Characterization of the Vitreous Proteome in Diabetes without Diabetic Retinopathy and Diabetes with Proliferative Diabetic Retinopathy

Cited by 215 publications

References 34 publications

(Pro)renin receptor is associated with angiogenic activity in proliferative diabetic retinopathy

(Pro)renin receptor is associated with angiogenic activity in proliferative diabetic retinopathy

Characterisation of the vitreous proteome in proliferative diabetic retinopathy

Dissection of Human Vitreous Body Elements for Proteomic Analysis

Contact Info

Product

Resources

About