Biology subjects

Schaefbauer, K.

Publications and source records attributed to Schaefbauer, K..

3 recordsLinked to original sources

bioRxiv2022

Blood Chemistry Analysis in an Active Pneumocystis Pneumonia (PCP) Model Treated with Selective CARD9 Inhibitor BRD5529

BackgroundPneumocystis pneumonia (PCP) in AIDS and other immunosuppressive states that result from absence of CD4 lymphocytic immunity, continues to be a significant cause of morbidity and mortality. We and others have shown the importance of CARD9 in PCP and other fungal infections, respectively. BRD5529 has been shown to be an effective in vitro and in vivo (18 hour) inhibitor of Pneumocystis {beta}-glucans induced proinflammatory response. These recent results, along with recent general safety and toxicology assessments suggests the application of BRD5529 in an active PCP mouse model of infection to assess initial blood toxicology parameters. MethodsTo assess preliminary blood toxicology, mice were injected intraperitoneally (IP) daily either with vehicle or BRD5529 at 1.0 mg/kg for one week starting at the 6th week of the PCP mouse model. After one week, mice were sacrificed, and blood collection postmortem was performed for blood chemistry analysis. ResultsAnalysis of blood chemistry showed a significant reduction in blood urea nitrogen (BUN) in the BRD5529 IP treated PCP mice cohort compared to the vehicle control group. All other blood chemistry parameters were not significantly different between the two groups. ConclusionsBRD5529 in this preliminary PCP treatment model displayed only significant changes in BUN levels in the BRD5529 treatment group versus the vehicle control group. All other blood chemistry parameters were statistically similar between the two groups.

immunology↗

bioRxiv2021

Preclinical and Toxicology Studies of BRD5529, a Selective Inhibitor of CARD9

BackgroundExuberant inflammation during Pneumocystis pneumonia leads to lung injury. CARD9 is a central mediator of inflammatory signaling mediated by C-type lectin receptors. CARD9 inhibitor BRD5529 has been shown to be an effective in vitro inhibitor of Pneumocystis {beta}-glucan-induced proinflammatory signaling and downstream TNF-alpha production, suggesting its viability as a candidate for preliminary drug testing as an anti- inflammatory agent in the rodent Pneumocystis pneumonia model (PCP). MethodsTo assess for potential toxicity, mice were injected intraperitoneally (IP) daily either with vehicle or BRD5529 at 0.1 mg/kg or 1.0 mg/kg for two weeks. Mouse weights were taken daily. At day 14, mice were euthanized, weighed, and analyzed by flexiVent for lung stiffness. Lungs, liver, and kidney were then harvested for H&E staining and pathology scoring. Lung samples were further analyzed for proinflammatory cytokines via ELISA and extracellular matrix generation via quantitative PCR (q-PCR). Blood collection postmortem was performed for blood chemistry analysis. ResultsBRD5529 at both doses of IP administration resulted in no significant changes in daily or final weight gain. Analysis of lung stiffness by flexiVent showed no significant differences between the control or treated groups. Furthermore, ELISA results for proinflammatory IL-1 Beta, IL-6, and TNF-alpha showed no major differences in the respective groups. qPCR analysis of extracellular matrix transcripts collagen type I, alpha 1 (Col1a1) and fibronectin (Fn) were statically similar as well in the treated and control groups. Examination and pathology scoring of H&E slides from lung, liver, and kidney from the each of the mice in all groups and subsequent pathology scoring showed no significant change. Blood chemistry analysis revealed similar, non-significant patterns. ConclusionsBRD5529 in our initial general safety and toxicology assessments displayed no inherent safety concerns in the analyzed parameters. These data support broader in vivo testing of the inhibitor as a timed adjunct therapy to the deleterious proinflammatory host immune response often associated with anti-Pneumocystis therapy.

pharmacology and toxicology↗

bioRxiv2019

The Vertebrate Codex Gene Breaking Protein Trap Library For Genomic Discovery and Disease Modeling Applications

The zebrafish is a powerful model to explore the molecular genetics and expression of the vertebrate genome. The gene break transposon (GBT) is a unique insertional mutagen that reports the expression of the tagged member of the proteome while generating Cre-revertible genetic alleles. This 1000+ locus collection represents novel codex expression data from the illuminated mRFP protein trap, with 36% and 87% of the cloned lines showcasing to our knowledge the first described expression of these genes at day 2 and day 4 of development, respectively. Analyses of 183 molecularly characterized loci indicate a rich mix of genes involved in diverse cellular processes from cell signaling to DNA repair. The mutagenicity of the GBT cassette is very high as assessed using both forward and reverse genetic approaches. Sampling over 150 lines for visible phenotypes after 5dpf shows a similar rate of discovery of embryonic phenotypes as ENU and retroviral mutagenesis. Furthermore, five cloned insertions were in loci with previously described phenotypes; embryos homozygous for each of the corresponding GBT alleles displayed strong loss of function phenotypes comparable to published mutants using other mutagenesis strategies (ryr1b, fras1, tnnt2a, edar and hmcn1). Using molecular assessment after positional cloning, to date nearly all alleles cause at least a 99+% knockdown of the tagged gene. Interestingly, over 35% of the cloned loci represent 68 mutants in zebrafish orthologs of human disease loci, including nervous, cardiovascular, endocrine, digestive, musculoskeletal, immune and integument systems. The GBT protein trapping system enabled the construction of a comprehensive protein codex including novel expression annotation, identifying new functional roles of the vertebrate genome and generating a diverse collection of potential models of human disease.

genomics↗