Simultaneous benzene, toluene, and xylene (BTX) exposure is a common phenomenon in the workplace and the environment, but has not been well defined by time-resolved molecular events leading to BTX-induced neurotoxicity in multicellular settings. To address these points, we derived an in vitro tri-culture system using SH-SY5Y with a supportive glial compartment (HMC3 + U87) and combined dose-dependent phenotypic profiling with time-resolved transcriptomic, proteomic and metabolic studies after 4, 12, 24, 36 and 48hours of BTX treatment. Working concentrations (IC10, IC20 and IC30) were determined at the end of an initial 24h dose-response step. Although BTX reduced cell viability in both monoculture and co-culture models, no significant differences in viability were observed between the two models at matched doses. Conversely, the co-culture model had increased sensitivity to sub-lethal toxic responses, which was evidenced by the higher levels of ROS and more obvious concentration-dependent responses to inflammatory, injury and the apoptosis-related markers. Transcriptional pathway dynamics were shown through time-course transcriptomics: initial enrichment of the cell cycle, DNA replication, and p53 signaling; mid-stage metabolic re-programming consisting of HIF-1 signaling, glycolysis/gluconeogenesis and pentose phosphate pathway; and later-stage enrichment of oxidative phosphorylation and Parkin pathways Time-course proteomics and metabolomics respectively indicated a temporal shift into mitochondrial energy dysfunction, proteostasis dysregulation, and neurodegeneration-associated modules. The integrative multi-omics analysis revealed oxidative phosphorylation, Parkinsonism, and thermogenesis as the convergent pathways. Additional evidence of early transcriptional compensation followed by a reduction of mitochondrial and neurofunctional proteins was obtained by time-resolved qPCR and western blot validation. Such results indicate a sequence of BTX neurotoxicity and provide a biologically meaningful multi-omics scheme to study mechanisms underlying and identify biomarkers.