- by Pattison, L. A., Dannawi, M., Smith, E. S. J.GPR65 is a proton-sensing G protein-coupled receptor implicated in inflammatory pain. In fibroblast-like synoviocytes (FLS), GPR65 activation promotes the release of proinflammatory cytokines capable of sensitizing sensory neurons. Following stimulation by protons, the synthetic agonist BTB09089, and the glycosphingolipid psychosine GPR65 undergoes internalization; however, the contribution of this trafficking to downstream signaling remains unclear. Using heterologous cell systems, the molecular mechanisms governing GPR65 internalization were first defined. Pharmacological and genetic inhibition of internalization revealed that intracellular trafficking is required for […]
- by Seymore, T., Hoffmann, S., Louro, P., Gardner, C., Goedken, M., Stapleton, P.Fetal health is heavily dictated by the maternal environment. Inhaling airborne pollutants, like particulate matter, is associated with pregnancy complications and fetal developmental pathologies, including fetal growth restriction (FGR). Because fetal growth is dependent on the placental transfer of nutrients from the maternal circulation, particularly glucose, investigating glucose transport capacity is critical to understanding the development of FGR associated with gestational inhalation of particulate matter. Pregnant Sprague Dawley rats were exposed to titanium dioxide nanoparticles (9.8 +/- 1.0 mg/m3) as […]
- by Eyal, S., Dallal, N., Rainish, A., Ziaikin, E., Malach, E., Niv, M. Y.Bitter taste receptors (TAS2Rs) are G- protein coupled receptors that detect chemically diverse compounds, including many clinically used drugs. TAS2R14 is expressed in many extraoral tissues and is activated by hundreds of ligands, including pharmaceutical drugs. Recent cryo-EM structures revealed a previously unrecognized intracellular binding pocket in TAS2R14, raising new questions regarding ligand binding modes. Here, we investigated the activation of TAS2R14 by Tamoxifen, Carbimazole, and Lidocaine using cell-based assays measuring proximal G-protein recruitment (BRET2) and downstream signaling (IP-One). Tamoxifen […]
- by Udoubom, I. A., Etim, O. E., Agu, G. E., Akpan, A. A., Jonah, U. I., James, E.- A. U., Patrick, I.Malaria remains one of the most pressing health problems, especially in Sub-Saharan Africa. Various antimalarial drugs, used to combat this debilitating illness, may directly or indirectly affect blood indices in humans. This study aims to evaluate the toxicological effects of sequential administration of Artemether-lumefantrine and sulfadoxine-pyrimethamine in male Wistar rats. Thirty (30) mature male Albino Wistar rats weighing between 190-280g were randomly divided into five groups comprising six (6) rats each. Group 1 served as control, Group 2 received Artemether-lumefantrine […]
- by Andres, J., Phengpol, N., Burmakin, M., Olauson, H., Patrakka, J., Moor, M. B.Acute kidney injury (AKI) affects millions of patients annually and is associated with high morbidity and mortality, to date no curative treatment exists. Drug-induced nephrotoxicity accounts for up to 25% of AKI cases, but individual susceptibility remains hard to predict. While genetic factors are suspected to play a part in this variability, the pharmacogenomics of nephrotoxin-induced kidney injury remain largely unknown. To investigate genetically determined susceptibility, we used precision-cut kidney slices (PCKS) from the two founder strains of the BXD […]
- by Polloni, L., Bieniasz, K. D., Gonteri, I., Frost, J. M.We present a generative machine learning pipeline for the design of linear antimicrobial peptides (AMPs). To extend diversity beyond synthetically validated peptide datasets ($sim$7,000 entries), we apply transfer learning by training a Generative Pre-trained Transformer (GPT) on the genomically derived AMPSphere dataset ($sim$863,000 entries), before fine-tuning on the Database of Antimicrobial Activity and Structure of Peptides (DBAASP). We assess the filtered sequences with a committee of Minimum Inhibitory Concentration (MIC) predictive models built with a Bi-LSTM architecture, and ESM-2 and […]
- by Jacobs, M. N., Kubickova, B., Person, E., Kamstra, J. H., Cabaton, N., Hoffmann, S., Jamin, A., Lacroix, M., Legler, J., Munic-Kos, V., Nijmeijer, S. M., Sinnige, T. L., Urien, L., Zalko, D.Cytochrome P450 (CYP) enzymes play a key role in the metabolism of both xenobiotics and endogenous compounds, and the activity of some CYP isoforms are susceptible to induction and/or inhibition by certain chemicals. As CYP induction and inhibition can significantly alter the in vivo fate of xenobiotics i.e., levels of parent chemicals and/or metabolites, and thus toxicity, CYP induction/inhibition data is needed for regulatory chemical toxicity hazard assessment. Utilizing available human in vivo pharmaceutical data, a successful validation was previously […]
- by Quartermain, E., Zhang, J., Marczylo, T., Gant, T. W., Jacobs, M. N.Cytochrome P450 (CYP)-mediated biotransformation of endogenous and xenobiotic substances can lead to altered exposure, toxicological impact, or adverse drug reactions. CYP induction data are fundamental to regulatory chemical toxicity hazard assessment because they directly affect the in vivo fate of xenobiotics, potentially influencing their safety and efficacy of pharmaceuticals, and impacting the safety assessment of industrial chemicals, and environmental contaminants. Here we report on the third laboratory supplementary validation of an established and previously validated human HepaRG TM in vitro […]
- by Etemadi, Y., Fields, T. A., Ramachandran, A., Jaeschke, H.Acetaminophen (APAP) overdose is the leading cause of acute liver failure (ALF), with acute kidney injury (AKI) contributing substantially to morbidity and mortality in those patients. To determine whether APAP-induced AKI depends on hepatic CYP2E1-mediated bioactivation, we used CYP2E1^flox/flox^ mice treated with AAV8-TBG-Cre to selectively delete hepatic CYP2E1 while preserving renal metabolism. Male and female mice received APAP (600 mg/kg) and were evaluated up to 48 hours for liver and kidney injury. Liver-specific CYP2E1 deletion reduced APAP hepatotoxicity, confirming the […]
- by Dall'Ara, E., sreenivasan, D., Oliviero, S., Boudiffa, M., Miller, R., Juarez, M., Bellantuono, I.Geroprotectors extend lifespan and improve several aspects of healthspan, yet their effects on skeletal ageing remain poorly understood. They hold potential advantages over current bone-targeted osteoporosis therapies, as they may simultaneously improve bone, neuromuscular function, and vision, thereby reducing the risk of falls, the major cause of fractures. Here we examined, for the first time, the long-term effects of rapamycin, acarbose, and 17-estradiol, administered at lifespan-extending doses on trabecular and cortical bone architecture in male and female UM-HET3 mice measured […]
- by Sharma, S., Tsang, Y. P., Unadkat, J. D.Pregnancy induces or represses hepatic drug metabolism. Whether pregnancy affects hepatic drug transport is unexplored. We previously showed that a cocktail of pregnancy-related hormones (PRHC) induces mRNA expression and activity of sodium/taurocholate cotransporting polypeptide (NTCP), organic anion transporter 2 (OAT2), and organic cation transporter 1 (OCT1, mRNA only) in differentiated HepaRG cells. Here, using HepaRG cells, we identified cortisol as the hormone primarily responsible for this induction and explored the underlying mechanisms. Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9-mediated knockdown […]
- by Kilbourn, E. A., Lowe, M. R., Panda, K., Bhaskaran, A., Zheng, G., Aalati, A. R., Malave, A., White, S., Graber, A., Zulkowski, N., Pepin, R., Salamova, A., Nemkov, T., D'Alessandro, A., Yadlapalli, S., Reddy, P., Meyhofer, E., Tennessen, J. M.Per- and polyfluoroalkyl substances (PFAS) are persistent environmental contaminants with poorly understood sublethal effects on insects. Perfluorooctanoic acid (PFOA), one of the most widely distributed legacy PFAS is increasingly recognized for altering organismal physiology beyond traditional toxicity endpoints. Here, we use the fruit fly Drosophila melanogaster as a model to examine how PFOA exposure during larval (juvenile) development reshapes insect life-history progression and metabolic homeostasis. Our studies reveal that at environmentally relevant concentrations (nM to low {micro}M), PFOA induces precocious […]
- by Sun, V. L., Wang, K., Kelly, E. J., Arnold, S. L. M.Transporters localized to the small intestinal epithelium govern the absorption of many orally administered drugs and nutrients, and thorough characterization of transporter protein abundance is key to accurate prediction of small molecule absorption. Previous studies have reported more than 100 solute carrier and ATP-binding cassette superfamilies expressed in the intestine, but only a small subset of these transporter proteins have been quantified in native intestinal tissue. In addition, a majority of published transporter abundance data for the intestine was collected […]
- by Bartoli, C., Anthony, A., Desetty, R.BackgroundThe CXCR2 receptor pathway plays a major role in inflammatory and invasive angiogenesis in human disease. ObjectiveWe evaluated AZD5069, a selective CXCR2 antagonist, as an angiogenesis inhibitor in human cell culture. MethodsHuman Umbilical Venous Endothelial Cells (HUVECs), Human Aortic Endothelial Cells (HAECs), and Human Pulmonary Artery Endothelial Cells (HPAECs) were cultured with standard in vitro techniques. AZD5069 (0, 8, 16, 32, 64, 128, 256 M) was evaluated as an angiogenesis inhibitor with fluorescent-labeled 5-Ethynyl-2-deoxyuridine (EdU) uptake to quantify endothelial cell […]
- by Kim, Y.-J., Woo, D. H.Mancozeb, a widely used fungicide composed of manganese ethylene-bis-dithiocarbamate with zinc salts, has raised concerns due to its potential neurotoxic effects. In this study, we investigated how chronic oral administration of mancozeb affects astrocyte function and neurobehavior in mice, focusing on store-operated Ca{superscript 2} entry (SOCE), mediated by Orai1 and STIM1. Mancozeb treatment at 0.5 {micro}g/kg/day for 4 weeks reduced glial fibrillary acidic protein (GFAP) expression in the hippocampus and corpus callosum of mice, indicating astrocyte atrophy. Further, administration at […]
- by Cai, D., Nguyen, H., Zhang, Y., Sharma, S., Schilke, A., Raychouni, R., Heredia, E., Abel-Santos, E., Firestine, S., Liu, W.Metabolic dysfunction-associated steatotic liver disease (MASLD) and Clostridioides difficile (C. difficile) infection (CDI) are clinically associated, yet there is limited effective treatment for both diseases. Bile salt analogs (BSAs) have demonstrated potential in treating either MASLD or CDI. We screened a library of BSAs (n=112) previously synthesized as potential inhibitors of C. difficile spore germination, for their therapeutic potential in reducing intracellular accumulation of fatty acids in HepG2 cells as candidates for prevention and treatment of both MASLD and CDI. […]
- by Abdel-Rahman, S., Gabr, M.The inhibitory microglial receptor LILRB4 (ILT3) suppresses amyloid-beta clearance in Alzheimers disease (AD) through ApoE-dependent signaling but remains undrugged by small molecules. Here, we report the AI-guided discovery of small molecule inhibitors that directly disrupt the LILRB4-ApoE interaction. Ultralarge-scale screening of [~]500 million compounds identified small molecules that bind LILRB4 with nanomolar affinity and competitively block ApoE engagement, as validated across orthogonal biophysical assays. Structural and mutational analyses define a tractable interdomain pocket that mediates ligand recognition. In human iPSC-derived […]
- by Shalaby, M.-A. W., Beeralingappa, N. C., Shrinidhi, A., Makafe, G. G., Nece, E., Patwardhan, A., Low-Beer, T., Kuki, A., Sheinerman, F., Weinrick, B., Flaherty, D. P., Chojnacki, M.New antitubercular agents acting through previously unexploited mechanisms are urgently needed. Using a drug-repurposing platform, we identified TI-374, a hydroxamic acid containing compound that inhibits Mycobacterium tuberculosis (Mtb) with sub-micromolar potency. Systems analysis, resistance mapping, supplementation assays, and biochemical studies showed that TI-374 inhibits two PLP-dependent aminotransferases, AlaA and HisC1. However, its activity is driven primarily by irreversible inhibition of AlaA, whereas HisC1 inhibition is only partially reversible, revealing differential reversibility between the two targets. Optimization yielded TI-801, a low-nanomolar […]
- by Coffin, D. J., Bhandari, S., Wittle, L. E., Ocius, K. L., Ongwae, G. M., Pires, M.While lipidation is a widely observed strategy to promote membrane permeation, whether the factors governing lipid-driven accumulation are shared across the divergent membranes of mammalian and Gram-negative cells remains unresolved. Here, we apply the Chloroalkane Azide-based Membrane Penetration (CHAMP) assay to a systematically designed library of lipid conjugates in both HeLa and E. coli cells. CHAMP, developed by our group, pairs a minimally disruptive azide tag with a cytosolically anchored HaloTag to quantify cytosolic accumulation directly. The two systems show […]
- by Abdel-Rahman, S., Gabr, M.Alzheimers disease (AD) remains a major unmet clinical challenge, with limited therapeutic strategies capable of effectively modulating neuroimmune dysfunction. Leukocyte immunoglobulin-like receptor B4 (LILRB4/ILT3) has recently emerged as an inhibitory microglial immune checkpoint implicated in ApoE-mediated suppression of amyloid-{beta} (A{beta}) clearance and inflammatory signaling, supporting its potential as a therapeutic target in AD. Here, we applied DNA-encoded library (DEL) screening of approximately 3.6 billion compounds to identify small molecule binders of LILRB4. Biophysical validation identified APX1 as a direct LILRB4 […]
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