Many veterans are burdened with chronic and complex medical conditions ranging from post-traumatic stress disorder, chronic pain, cancer, obesity, pre-diabetes mellitus, frank diabetes, acute kidney injury, chronic kidney disease, end-stage renal disease, to a host of sclerotic disorders. This application is for a Cytation 5 Cell Imaging Multi-Mode Reader Imaging System with connected BioSpa 8 Automated Incubator and Robotic Arm that will be used in experiments addressing the following Aims of Major Users toward combating these health issues: 1. Karin Westlund High, Ph.D. ?Modulating Pain Generators of Chronic Trigeminal Neuropathic Pain?25% Test efficacy of the extended release NA?1R antagonist doxazosin for reduction of chronic pain/anxiety/ depression related behaviors. Chronic pain has poor therapeutic response to analgesics, especially opioids, and remains a significant challenge. Veteran patients report more severe chronic pain than non-veteran patients and are twice as likely to die of an opioid overdose in attempts to find relief. There is an urgent need to identify chronic pain induced brain RNA/protein expression signatures to assist in the development of non-opioid pharmaceutical compounds. The proposed studies are to determine efficacy of the extended release NA?1R antagonist doxazosin when tested in weeks 8-10. Automated imaging of histological samples with the requested imaging system will assess cellular and biochemical changes along the pain neuraxis and significantly facilitate experimental progress. 2. Jenny Mao, M.D. ?Combinations of Grape Seed and Milk Thistle Extracts Against Lung Cancer? 25% Determine the maximum tolerated dose (MTD), PK/PD, and anti-cancer effects of leucoselect phytosome (LP) with siliphos in murine models. Historically, the VA has emphasized oncology research, particularly finding new treatments. Goal is to systematically determine the synergistic, antineoplastic properties of combinations of a standardized grape seed procyanidin extract (GSE), leucoselect phytosome (LP), and milk thistle silymarin extract (MTE), siliphos, against lung cancer using both in vitro and in vivo preclinical models. We hypothesize that oral administration of LP and siliphos will synergistically inhibit cancer growth and induce apoptosis in human lung cancer xenograft models, via favorable modulations of mechanisms associated with lung tumorigenesis/promotion. Effects of varying doses on proliferation and apoptosis in co-cultured cells over time will be determined and correlated to levels of GSE, MTE, and metabolites. The Cytation 5 Imaging System will enable automated repeated imaging of cultured cells with different treatments for PK/PD studies. 3. Manoocher Soleimani, M.D. ?The Role of Collecting Duct Chloride Transporters in Salt Absorption and Blood Pressure Momeostasis? 25%. Ascertain the role of chloride transporter KBAT in salt absorption in the collecting duct in pathophysiologic states. Many veterans are admitted to the hospital with altered systemic blood pressure or excess fluid. The kidney?s distal nephron of the kidney plays a central role in vascular volume homeostasis and blood pressure regulation by regulating the absorption of salt via specialized molecules. We hypothesize that kidney brain anion transporter (KBAT) plays an important role in salt absorption in the entire collecting duct following increased delivery of salt consequent to the inactivation/inhibition of salt absorbing transporter proteins or in response to increased dietary salt intake. Using a combination of genetically engineered mouse models, metabolic studies, blood pressure recording, and molecular studies, we will determine the role of KBAT in salt absorption in pathophysiologic states. The requested imaging system will accelerate acquisition and analysis of transporter protein expression in kidney sections under different experimental conditions. Projects of Minor Users will monitor mitophagy through colocalization of mitochondrial probes with autophagy biomarkers, quantify cellular and molecular mechanisms that lead to gastrointestinal dysbiosis, hospital-acquired fungal infections, chronic kidney disease and opioid use disorder using the Cytation 5 Imaging System.
Many veterans of the United States of America suffer from chronic and complex health problems including chronic pain, cancer, obesity, diabetes, heart, kidney, and skin diseases. The shared equipment requested in this application, the automated Cytation 5 Cell Imaging Multi-Mode Reader with connected BioSpa 8 Automated Incubator and Robotic Arm, will enhance and facilitate experiments currently being undertaken to address chronic pain, cancer treatments, safety of diagnostic drugs and the impact of sex on toxicity, Gulf War syndrome, bacterial infections, and irritable bowel syndrome. The discoveries, using automated, high-throughput 21st Century technology, will serve as the basis to design new and improved ways to serve veteran patients.