Background/Purpose: Kawasaki disease (KD), a predominantly coronary vasculitis of childhood, remains the most common cause of acquired heart disease of childhood in the developed world, and the developing concerns of myocardial damage in the form of cardiac fibrosis amplifies the urgency for a greater understanding both in terms of management and mechanism of this illness. At present, no biomarkers exist for the disease, response to therapy, persistence of inflammation or long term fibrosis; similarly, our understanding of the overall mechanism of the arterial remodeling and myocardial damage in KD is limited. Microwave and magnetic (M²) proteomics is a method whereby accelerated processing of proteins allows for high-throughput sample preparation and isolation of peptides for mass spectrometry. Our hypothesis was that protein biomarkers for vascular and myocardial inflammation would be identifiable in cardiac tissue from mice with a model of KD, that these markers would show response to a therapy such as interleukin-1 (IL-1) inhibition, offering insight into disease mechanisms.

Methods: KD was induced via the established model of lactobacillus casei cell wall extract (LCWE) injection in 4-6 week old male mice. Groups of mice either injected with LCWE alone, LCWE and anakinra, or saline for normal controls. After 2 weeks, mice were sacrificed and aortic root region of the heart was extracted, homogenized and processed. Tissue then underwent mass spectrometry analysis. Probability-based protein database searching of spectra was performed. Data was analyzed for pathway enrichment using STRING and REACTOME pathway analysis.

Results: Several proteins demonstrated multifold elevation in diseased animals as compared to controls, with certain proteins with primarily skeletal muscle and cardiac muscle expression notably elevated and down expressed after therapy. Proteomic analysis demonstrated clustering of diseased (KD) animals apart from controls and anakinra treated animals, which clustered together when analyzed by k-means testing. Preliminary pathway analysis suggests that in KD diseased hearts, there is enrichment for proteins involved in platelet aggregation and activation (p<0.01) and cardiac epithelial to mesenchymal transformation (p=0.04). Anakinra treated animals demonstrate enrichment for wnt signaling and downstream wnt signaling effectors beta-catenin and T cell factor (TCF) (p=0.01).

Conclusion: Potential biomarker proteins for disease activity and response to therapy were found based on the hypothesis of differential protein expression therapy and disease. These proteins require further validation. Overall, anakinra treated animals had a cardiac proteome that was more similar to normal controls than to KD mice. Preliminary pathway analysis suggested pathways for vascular injury and intimal hyperplasia in diseased animals, and induction of a regulatory immune phenotype mediated by wnt/beta-catenin in anakinra treated animals.

« Back to 2018 ACR/ARHP Annual Meeting

ACR Meeting Abstracts - https://acrabstracts.org/abstract/microwave-and-magnetic-m2-myocardial-proteomics-in-the-mouse-model-of-kawasaki-disease-demonstrates-normalization-of-the-proteome-after-interleukin-1-inhibition-potential-novel-biomarkers-and-sugg/