Hypoxia-induced carbonic anhydrase mediated dorsal horn neuron activation and induction of neuropathic pain.

ABSTRACT: Neuropathic pain, such as that seen in diabetes mellitus, results in part from central sensitisation in the dorsal horn. However, the mechanisms responsible for such sensitisation remain unclear. There is evidence that disturbances in the integrity of the spinal vascular network can be causative factors in the development of neuropathic pain. Here we show that reduced blood flow and vascularity of the dorsal horn leads to the onset of neuropathic pain. Using rodent models (type 1 diabetes and an inducible endothelial-specific vascular endothelial growth factor receptor 2 knockout mouse) that result in degeneration of the endothelium in the dorsal horn, we show that spinal cord vasculopathy results in nociceptive behavioural hypersensitivity. This also results in increased hypoxia in dorsal horn neurons, depicted by increased expression of hypoxia markers such as hypoxia inducible factor 1α, glucose transporter 3, and carbonic anhydrase 7. Furthermore, inducing hypoxia through intrathecal delivery of dimethyloxalylglycine leads to the activation of dorsal horn neurons as well as mechanical and thermal hypersensitivity. This shows that hypoxic signalling induced by reduced vascularity results in increased hypersensitivity and pain. Inhibition of carbonic anhydrase activity, through intraperitoneal injection of acetazolamide, inhibited hypoxia-induced pain behaviours. This investigation demonstrates that induction of a hypoxic microenvironment in the dorsal horn, as occurs in diabetes, is an integral process by which neurons are activated to initiate neuropathic pain states. This leads to the conjecture that reversing hypoxia by improving spinal cord microvascular blood flow could reverse or prevent neuropathic pain.

Quantifying Vascular Remodeling in the Mouse Spinal Cord.

The spinal cord, a compartment of the central nervous system, is made up of a number of architecturally distinct neural centers that influence an array of neurophysiological systems. The primary role of the spinal cord is the modulation of sensory and motor function by acting as a relay station between the periphery and the brain. Inherently these are considered as neural networks, however the functional dynamics of these tissues consist of a heterogenic population of cell types, all working in harmony to maintain physiological function. Part of this cellular diversity comprises of the vascular network that delivers essential nutrients and oxygen to the spinal cord tissue, whilst also protecting it from potentially tissue damaging substances such as foreign entities including toxic pharmacological agents or pathogens. The viability of the spinal cord is dependent upon the harmonious balance between opposing angiogenic processes; vascular remodeling and vascular regression, tipping the balance to either side contributes to neurodegeneration. Exploring vascular remodeling in the central nervous system requires consideration of the anatomical landscape of the spinal cord and the dynamic nature of the microvasculature. Utilizing immunofluorescent staining and 3D image rendering analysis of the endothelium and mural cell population allows for investigation of cellular as well as molecular mediation of vascular remodeling in the spinal cord. This method can be utilized in a range of rodent models (utilizing pharmacological, disease models, transgenic and/or viral approaches) offering extensive appreciation of the blood-spinal cord barrier.

LRG1 as a novel therapeutic target in eye disease.

Retinal and choroidal diseases are major causes of blindness and visual impairment in the developed world and on the rise due to an ageing population and diabetes epidemic. Standard of care is centred around blockade of vascular endothelial growth factor (VEGF), but despite having halved the number of patients losing sight, a high rate of patient non-response and loss of efficacy over time are key challenges. Dysregulation of vascular homoeostasis, coupled with fibrosis and inflammation, are major culprits driving sight-threatening eye diseases. Improving our knowledge of these pathological processes should inform the development of new drugs to address the current clinical challenges for patients. Leucine-rich α-2 glycoprotein 1 (LRG1) is an emerging key player in vascular dysfunction, inflammation and fibrosis. Under physiological conditions, LRG1 is constitutively expressed by the liver and granulocytes, but little is known about its normal biological function. In pathological scenarios, such as diabetic retinopathy (DR) and neovascular age-related macular degeneration (nvAMD), its expression is ectopically upregulated and it acquires a much better understood pathogenic role. Context-dependent modulation of the transforming growth-factor ß (TGFß) pathway is one of the main activities of LRG1, but additional roles have recently been emerging. This review aims to highlight the clinical and pre-clinical evidence for the pathogenic contribution of LRG1 to vascular retinopathies, as well as extrapolate from other diseases, functions which may be relevant to eye disease. Finally, we will provide a current update on the development of anti-LRG1 therapies for the treatment of nvAMD.