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A novel molecular imaging for early diagnosis of diabetic retinopathy

Session Details

Session Title: FP-15 Vascular Diseases and Diabetic Retinopathy V

Session Date/Time: Sunday 14/09/2014 | 08:00-10:00

Paper Time: 09:20

Venue: Boulevard D

First Author: : D.Sun CHINA

Co Author(s): :                  

Abstract Details

Purpose:

The incidence of Diabetic retinopathy (DR), a microvascular manifestation of diabetes, is worldwide rapidly rising. Biomarkers of DR are urgently needed, as vision loss can be prevented with early diagnosis and treatment. Here, we introduce a novel biomarker of early diabetic micro-vasculopathy. The dynamics of growth factor expression is of paramount interest. Using our unique molecular imaging approach, we investigate early endothelial changes in retinas of living diabetic animals.

Setting:

the 2nd affiliated Hospital of Harbin medical university

Methods:

Molecular imaging probes were custom made by covalently conjugating carboxylated fluorescent microspheres (MSs, 2μm) with mouse IgG, anti-VEGFR-2, or VEGF. MSs (6x108) were injected into the tail vein of each animal. MS interaction in the fundus vessels was studied using a scanning laser ophthalmoscope (SLO, HRA2). 30min after MS injection, rats were perfused with PBS and rhodamine-labeled conA. Flatmounts were prepared for ex vivo evaluation of accumulated MS and leukocytes. Western blotting was used to quantify the amount of VEGFR-2 in retinal vessels of normal and diabetic animals. Diabetes was induced by intraperitoneal injection of Streptozotocin in male Long-Evans rats (180-200g).

Results:

The number of a-VEGFR-2-Ab and VEGF molecules on our imaging probes was quantified by flow-cytometry, to be 27453 and 27094, respectively. Western blot analysis showed significantly higher VEGFR-2 expression in diabetic retinas (P=0.04). Immunohistochemistry showed no VEGFR-2 staining in normal retinal vessels, while retinal capillaries were distinctively positive for VEGFR-2. In vivo molecular imaging showed significantly higher a-VEGFR-2-Ab-conjugated imaging probes in diabetic retinas (P=0.02), compared to normal controls. Nearly all VEGFR-2 signal was detected in retinal capillaries.

Conclusions:

The present work introduces a novel biomarker of early diabetic microvasculopathy. The pattern and dynamic of VEGFR-2 in diabetic retinal capillaries might herald the progression of the diseases into the proliferative stage. Our molecular imaging approach to in vivo VEGFR-2 detection could be further developed to detect subclinical signs of diabetic retinopathy in humans, impacting clinical management of DR.

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