MULTI-OMICS CORE C SUMMARY This PPG is addressing an important hypothesis that early life stress (ELS) leads to reprogramming throughout the body resulting in an increased risk of developing CVD in adulthood. The mechanistic links between ELS and CVD and resilience in adulthood is not established. Epigenetic regulation of transcription or the gut microbiome and microbial-derived factors are hypothesized to be sensitive to ELS and may be modifiable to mediate resiliency. The primary goal and function of the Multi-omics Core (Core C) is to provide start-of-the-art processing of samples for determination of transcriptomes, DNA methylomes, DNA chromatin accessibility, metabolomes, and microbiomes required by all Projects. The objective of the Core is to provide consistency and rigor in the multi-omics experiments and provide innovative technologies to meet the goals of the Projects. The specific aims of Core C are: Aim 1 Transcriptomic and epigenomic determination. The epigenome is an important connection between the environment and the genes expressed in each cell (transcriptomes) and it is regulated by DNA methylation and chromatin structure (accessibility). Core C will facilitate determination of transcriptomes, and epigenome global DNA methylation and chromatin accessibility for the Projects. Aim 2: Microbiome and metabolite profiling. In this PPG, it is hypothesized that ELS results in gut microbial diversity changes that result in significant alterations to circulating short chain fatty acids (SCFA) or other metabolites. Core C will prepare samples for microbiome 16S sequencing for identifying and quantifying microbial diversity. The Core will also prepare samples for SCFA and untargeted metabolite determination by mass spectrometry. Aim 3: Evaluating new technologies, optimization while promoting fiscal responsibility. In this era of multi-omics, technologies are rapidly evolving. Thus, to provide the Projects with the latest technologies to test their hypotheses, Core C will evaluate new technological platforms. Core C will optimize all protocols for sample preparation for the multi-omics approaches listed above. By centralizing the mutli-omic sample preparation in Core C, we will purchase kits and reagents in bulk leading to substantial cost savings. Core C will interact directly with all Projects, receive clinical samples from Core B, and work directly with Core A for sample identification, curating, data deposition and analyses, and data management. Core A will complete all bioinformatics and biostatistical analyses for data generated in Core C. The approaches used in Core C ensure that the PPG is using the latest technologies for determining mechanisms of ELS induced cardiovascular disease risk with a bench to beside approach.