Abstract TMP81: Immunocompetent Neural Organoids Respond To Blood Stimulation: A Novel Model Of Intracerebral Hemorrhage

Abstract

Introduction: The lack of treatments for intracerebral hemorrhage (ICH) suggests the need for novel preclinical models of this disease. To that end, this study aimed to explore the utility of an ICH neural organoid model. Methods: Human neural organoids consisting of several cell types, including microglia, were provided by Stem Pharm, Inc. On DIV 21, organoids were treated with human whole blood (n=3 organoids in each condition) for 24 hours. To serve as positive and negative controls, one organoid was treated with lipopolysaccharide (LPS) for 24 hours and three remained untreated. Human IL-6 concentrations were measured in the culture media prior to and after treatments using an enzyme-linked immunosorbent assay (ELISA) and the fold-change from baseline was normalized to the negative control condition. A one-way ANOVA with Dunnett’s test was then performed to compare IL-6 secretion between the control and treatment conditions. Results: LPS and blood stimulation both induced IL-6 secretion after 24 hours. The greatest fold increase in IL-6 concentration relative to the control group was observed with 10% blood stimulation (29.76 +/- 7.91, p<0.0001) followed by LPS (23.43 +/- 11.18, p<0.001) and 5% blood (22.56 +/- 7.91, p<0.0001). Significant fold increases in IL-6 secretion were also observed in the lower concentration conditions of 2.5% (15.75 +/- 7.91, p<0.001), 1.25% (11.46 +/- 7.91, p<0.01), and 0.6% blood (11.60 +/- 7.91, p<0.01). Conclusion: This preliminary study is the first to explore a neural organoid model of ICH. Comparable to LPS, the observed trends in IL-6 secretion suggest that whole blood is a potent, dose-dependent inducer of organoid inflammation. Given the blood-induced neuroinflammation that occurs during ICH, these initial results support a potential role for neural organoids in ICH modeling. Future work will aim to further characterize the organoid inflammatory response and validate this ICH model system via gene expression analyses.