Normalisation of vascular microenvironment through improvement of neuroinflammation in diabetic retinopathy
Diabetes is an increasingly prevalent condition globally and prevalence is expected to rise from an estimated 366 million people in 2011 to 522 million by 2030. The prevalence of diabetic retinopathy in diabetes patients is 22-37% and it is one of the leading causes of blindness in developed countries. This project focuses on the causal links between neuroinflammation and microvasculopathy in the diabetic retina, particularly through glutamate excitotoxicity and interrogation of whether neuropeptides (SP and CGRP) in combination with an anti-inflammatory agent (minocyclines) can normalise vascular microvascular environment and ameliorate diabetic retinopathy.
We characterize structural and functional neurodegeneration in diabetic neuropathy including expression of neuropeptides, perivascular sensory nerve defects and apoptosis of neural cells by means of qPCR, ELISA, immunostaining, TUNEL assay and microelectrode array using both human donor retina and streptozotocin-induced diabetic mice retina.
The effect of glutamate, advanced glycation end products and hyperglycemic insult on perivascular sensory nerves, retinal inflammation and microvessel integrity is interrogated via intravitreal injections in a murine model prior to tissue harvesting for qPCR and immunostaining. In vitro, mouse aortic ring assay, HUVEC tubule formation assay, choroidal and retina explant culture with different treatment will be tested.
To investigate mobilisation and homing of EPC´s, cells are isolated and purified from bone marrow by magnetic separation before in vitro transwell migration assays, and in vivo cell labelling and trafficking studies are performed in diabetic mice.
Finally, SP, CGRP, anti-glutamate receptor and minocycline are delivered locally or systemically to evaluate their possible therapeutic efficacy in diabetic retinopathy with respect to reduction of neuroinflammation and reduction of microvascular abnormality.