TRP-2 / gp100 DNA vaccine and PD-1 checkpoint blockade combination for the treatment of intracranial tumors.

Intracranial tumors present a significant therapeutic challenge due to their physiological location. Immunotherapy presents an attractive method for targeting these intracranial tumors due to relatively low toxicity and tumor specificity. Here we show that SCIB1, a TRP-2 and gp100 directed ImmunoBody® DNA vaccine, generates a strong TRP-2 specific immune response, as demonstrated by the high number of TRP2-specific IFNγ spots produced and the detection of a significant number of pentamer positive T cells in the spleen of vaccinated mice. Furthermore, vaccine-induced T cells were able to recognize and kill B16HHDII/DR1 cells after a short in vitro culture. Having found that glioblastoma multiforme (GBM) expresses significant levels of PD-L1 and IDO1, with PD-L1 correlating with poorer survival in patients with the mesenchymal subtype of GBM, we decided to combine SCIB1 ImmunoBody® with PD-1 immune checkpoint blockade to treat mice harboring intracranial tumors expressing TRP-2 and gp100. Time-to-death was significantly prolonged, and this correlated with increased CD4+ and CD8+ T cell infiltration in the tissue microenvironment (TME). However, in addition to PD-L1 and IDO, the GBM TME was found to contain a significant number of immunoregulatory T (Treg) cell-associated transcripts, and the presence of such cells is likely to significantly affect clinical outcome unless also tackled.

The Role of Vascular-Immune Interactions in Modulating Chemotherapy Induced Neuropathic Pain.

Chemotherapy causes sensory disturbances in cancer patients that results in neuropathies and pain. As cancer survivorships has dramatically increased over the past 10 years, pain management of these patients is becoming clinically more important. Current analgesic strategies are mainly ineffective and long-term use is associated with severe side effects. The issue being that common analgesic strategies are based on ubiquitous pain mediator pathways, so when applied to clinically diverse neuropathic pain and neurological conditions, are unsuccessful. This is principally due to the lack of understanding of the driving forces that lead to chemotherapy induced neuropathies. It is well documented that chemotherapy causes sensory neurodegeneration through axonal atrophy and intraepidermal fibre degeneration causing alterations in pain perception. Despite the neuropathological alterations associated with chemotherapy-induced neuropathic pain being extensively researched, underlying causes remain elusive. Resent evidence from patient and rodent studies have indicated a prominent inflammatory cell component in the peripheral sensory nervous system in effected areas post chemotherapeutic treatment. This is accompanied by modulation of auxiliary cells of the dorsal root ganglia sensory neurons such as activation of satellite glia and capillary dysfunction. The presence of a neuroinflammatory component was supported by transcriptomic analysis of dorsal root ganglia taken from mice treated with common chemotherapy agents. With key inflammatory mediators identified, having potent immunoregulatory effects that directly influences nociception. We aim to evaluate the current understanding of these immune-neuronal interactions across different cancer therapy drug classes. In the belief this may lead to better pain management approaches for cancer survivors.

The consequence of endothelial remodelling on the blood spinal cord barrier and nociception.

Nociception is a fundamental acute protective mechanism that prevents harm to an organism. Understanding the integral processes that control nociceptive processing are fundamental to our appreciation of which cellular and molecular features underlie this process. There is an extensive understanding of how sensory neurons interpret differing sensory modalities and intensities. However, it is widely appreciated that the sensory neurons do not act alone. These work in harmony with inflammatory and vascular systems to modulate pain perception. The spinal cord has an extensive interaction with the capillary network in the form of a blood spinal cord barrier to ensure homeostatic control of the spinal cord neuron milieu. However, there is an extensive appreciation that disturbances in the blood spinal cord barrier contribute to the onset of chronic pain. Enhanced vascular permeability and impaired blood perfusion have both been highlighted as contributors to chronic pain manifestation. Here, we discuss the evidence that demonstrates alterations in the blood spinal cord barrier influences nociceptive processing and perception of pain.

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.