Abstract 3102: NELISA High-throughput Proteomics Captures Disease Phenotypes And Cytokine Interactions By Quantifying Immune, Metabolic, And Cardiovascular Markers

The variety of cell types and interactions involved in vascular diseases pose a significant challenge to quantifying the breadth of inflammatory, metabolic, and cardiovascular proteins that could provide mechanistic information or serve as clinical biomarkers, particularly at the scale needed for population studies or drug discovery screens. Unfortunately, proteomics tools able to quantify these proteins compromise either content or throughput/cost, limiting their use. To overcome this issue, we developed the nELISA: a high-throughput miniaturized ELISA capable of quantifying 272 cytokines, adipokines, proteases, soluble receptors, and growth factors, at 10x-reduced cost compared to previous tools. We applied the nELISA to model systems as a proof-of-concept for use in high-throughput screens. We ran the largest PBMC secretome screen to date, in which ~10,000 PBMC samples were profiled at a throughput of 1536 samples/day. Cells were treated with a selection of myeloid and T cell stimuli to model different disease states, and were further perturbed with a selected library of 80 recombinant cytokine perturbagens; secreted proteins were profiled in all samples, resulting in ~2M datapoints. In addition to distinguishing the various disease states, the nELISA captured 449 cytokine interactions, where addition of a specific perturbagen resulted in a change in the expression levels of a secreted protein. Of note, we found 3.2-fold more of these interactions than reported at the transcriptomic level from the CytoSig database. By clustering these cytokine interactions, we identified perturbagens with shared mechanisms of action, including results supporting the use of IL-1 Receptor antagonist to replace IFN beta in certain indications. We will expand on these findings and discuss how the nELISA is being applied to HUVEC cultures, whole blood assays, and patient plasma/serum.