Faculty Bibliography

Emerging evidence links the receptor for advanced glycation endproducts (RAGEs) to the pathogenesis of tissue damage in chronic metabolic and inflammatory diseases. In human subjects, multiple reports suggest that in the plasma/serum, circulating levels of distinct forms of soluble RAGEs may be biomarkers of the presence or absence, and the extent of chronic disease. These considerations prompt us to consider in this review, what are soluble RAGEs; how are they formed; what might be their natural functions; and may they serve as biomarkers of inflammatory and metabolic disease activity? In this brief review, we seek to address what is known and suggest new areas for scientific investigation to uncover the biology of soluble RAGEs.

Types 1 and 2 diabetes are on the rise worldwide. Although the treatment of hyperglycemia has benefitted from recent advances, aggressive efforts to maintain euglycemia may be fraught with risk, especially in older subjects or in subjects vulnerable to hypoglycemic unawareness. Hence, strategies to prevent and treat the complications of hyperglycemia are essential. In this review, we summarize recent updates on the biology of the receptor for advanced glycation endproducts (RAGE) in the pathogenesis of both micro- and macrovascular complications of diabetes, insights from the study of mouse models of obesity and diabetic complications, and from associative studies in human subjects. The study of the mechanisms and consequences of the interaction of the RAGE cytoplasmic domain with the formin, mDia1, in RAGE signal transduction, will be discussed. Lastly, we review the 'state-of-the-art' on RAGE-directed therapeutics. Tackling RAGE/mDia1 may identify a novel class of therapeutics preventing diabetes and its complications

Background: The risk of developing Alzheimer's disease (AD) is magnified in individuals with metabolic dysfunction, specifically obesity and type 2 diabetes. In cases of insulin deficiency or resistance, elevated and fluctuating levels of blood glucose lead to the production of Advanced Glycation End Products (AGEs) that bind their receptor, RAGE, with pathological consequences. AGE-RAGE ligand binding induces intracellular signaling cascades, in part via the formin signal transduction effector, diaphanous- 1 (mDia1). Activation of this cellular mechanism activates NF-KB activation, upregulates pro-inflammatory molecules, increases RAGE expression, and ignites a positive feedback loop driving chronic inflammation in the periphery. Given that AGEs are increased in the brains of both diabetic and AD patients, here we investigate AGE-RAGE binding and subsequent mDia1 signal transduction as a possible mechanism of neuroinflammation, which contributes to the pathogenesis of AD. Methods: mDia1 and RAGE expression in microglia was evaluated by Immunoflourescent IHC in temporal cortex brain slices of AD and Non-Demented Aged human tissue. CD11b+ microglia were isolated from young (<10 month) and old (>16 month) APP London mice and aged matched controls and subjected to molecular analysis. BV-2 microglial- like cells were stimulated by the prototypic RAGE ligand, Carboxy Methyl Lysine (CML-AGE) (100 mug/mL) for 24 h, harvested, and Ager (RAGE), Drf1 (mDia1), Cd36, and Cd68RNA transcript levels were analyzed by q-RT-PCR. Results: mDia1 and RAGE are highly expressed in human AD and aged brain and colocalize, at least in part, to CD68+ activated microglia. mDia1 is highly expressed in CD11b+ microglia from APP London mice vs. aged matched controls. CML-AGE stimulated BV-2 cells display a 2-fold increased expression of mDia1, RAGE, CD36, and CD68 vs. vehicle treatment. Conclusions: RAGE and mDia1 are highly expressed in AD human brain, supporting their possible role in mediating pathogenesis. Acute stimulation with RAGE ligands upregulates inflammatory and phagocytosis markers in cultured BV-2 cells. Primary isolates of murine CD11b+ microglia demonstrate high levels of expression of mDia. These data suggest that RAGE may exert its pathogenic effects in AD brain, at least in part via mDia1-mediated neuroinflammation, thereby driving the AD phenotype. (Figure presented)

We investigated treatment with a receptor for advanced glycation endproduct (RAGE) blocking antibody on angiogenic response to hind limb ischemia in diabetic mice. Streptozotocin treated C57BL/6 mice received either murine monoclonal anti-RAGE F(ab')2 intraperitoneally (n=10) or saline (n=9) for 9 weeks. Diabetic plus 10 non-diabetic C57BL/6 mice underwent left femoral artery ligation and 5 days later angiogenesis imaging with (99m)Tc-Arg-Gly-Asp (RGD) nanoSPECT/CT. Twenty-four days later, hind limb blood flow was measured with ultrasound, the mice were euthanized, and tissue was taken for immunohistochemistry. The angiogenic imaging signal in ischemic limbs was higher in RAGE-ab treated versus saline treated mice at day 5 (3.1+/-1.4 vs 1.68+/-0.35, p=0.02) and blood flow was higher at day 24 (1.49+/-0.5 vs 0.61+/-0.39, p=0.04). Immunohistochemistry of ischemic muscles showed greater capillary density in the RAGE-ab treated group versus the vehicle-treated group (p<0.001) (NS from non-diabetic mice). In conclusion, treatment with anti-RAGE F(ab')2 in diabetic mice improves neovascularization in the ischemic leg.

SCOPE: Advanced glycation end-products (AGEs) are endogenously produced and are present in food. N(epsilon) -carboxymethyllysine (CML) is an endothelial activator via the receptor for AGEs (RAGEs) and is a major dietary AGE. This work investigated the effects of a CML-enriched diet and RAGE involvement in aortic aging in mice. METHODS AND RESULTS: After 9 months of a control diet or CML-enriched diets (50, 100, or 200 mugCML /g of food), endothelium-dependent relaxation, RAGE, vascular cell adhesion molecule-1, and sirtuin-1 expression, pulse wave velocity and elastin disruption were measured in aortas of wild-type or RAGE(-/-) male C57BL/6 mice. Compared to the control diet, endothelium-dependent relaxation was reduced in the wild-type mice fed the CML-enriched diet (200 mugCML /g) (66.8 +/- 12.26 vs. 94.3 +/- 2.6%, p < 0.01). RAGE and vascular cell adhesion molecule-1 (p < 0.05) expression were increased in the aortic wall. RAGE(-/-) mice were protected against CML-enriched diet-induced endothelial dysfunction. Compared to control diet, the CML-enriched diet (200 mugCML /g) increased the aortic pulse wave velocity (86.6 +/- 41.1 vs. 251.4 +/- 41.1 cm/s, p < 0.05) in wild-type animals. Elastin disruption was found to a greater extent in the CML-fed mice (p < 0.05). RAGE(-/-) mice fed the CML-enriched diet were protected from aortic stiffening. CONCLUSION: Chronic CML ingestion induced endothelial dysfunction, arterial stiffness and aging in a RAGE-dependent manner.