Transcriptomic Profile of Neuroinflammation in Irradiated Brain Following C5aR1 Inhibition, With and Without Brain Tumor

Expression of 757 genes related to neuroinflammation was analyzed across 12 treatment groups of male mice to investigate the neuroprotective effects of C5a Receptor 1 (C5aR1) inhibition against cranial irradiation for brain cancer therapy, utilizing both a pharmacologic (PMX205) and genetic (C5aR1-KO) approach. Samples from 58 male mice, ages 23-24 weeks at time of perfusion, were analyzed using the commercially available nCounter® Mouse Neuroinflammation Panel from NanoString™, with 3-4 samples for each of the 15 treatment groups. For the pharmacologic model, C57BL/6J mice were randomly assigned to treatments groups with the following parameters: with or without brain tumor (astrocytoma, CT2A cell line, or glioblastoma, GL261 cell line); with or without single-dose cranial irradiation (9 Gy vs 0 Gy); and with or without C5aR1 inhibitor drug treatment (PMX205, 1mg/kg body weight injected subcutaneously for 7 days and 100 µg daily in drinking water for 30 days). For the transgenic model, C5aR1 KO mice were randomly assigned to the following treatment groups: with or without brain tumor (astrocytoma, CT2A cell line); and with or without single-dose cranial irradiation (9 Gy vs 0 Gy). Note: in the treatment group C5aR1 KO + 0 Gy, samples “PMX 25a” and “PMX25b”, as well as “PMX26a” and “PMX26b”, are each pairs of technical replicates, for a total of 2 biological replicates for this group; all other treatment groups contain 4 biological replicates. Sample ID’s “PMX60” thru “PMX71” originate from a glioblastoma model and were run separately from Sample ID’s “PMX13” thru “PMX59.” Both sample sets underwent the same protocols described below.