Understanding the Interaction between Type 2 Diabetes and COVID-via Extracellular Vesicle Proteomic and Phosphoproteomic Signatures

ABSTRACT

Evidence supporting the involvement of EVs in the pathogenesis/severity of SARS-CoV-2 infection is starting to accumulate. However, little is known about their specific associations in the context of COVID-and type 2 diabetes interaction. Our study included 48 plasma samples (N=12/group) obtained from COVID-patients with and without diabetes and from patients with non-COVID-acute respiratory diagnosis (RSP) with and without diabetes. Participants were identified from a set of 494 patients hospitalized at AdventHealth in June-August 2020. Important efforts were made to ensure the homogeneity of the study cohort. Patients with type 1 diabetes, or pregnant, or that went directly into the ICU were excluded, and 4 balanced groups were identified after 10,000 random cohorts were generated and differences in age, gender, race, and ethnicity statistically assessed. EVs were isolated using EVTRAP (Tymora) . Mass spectrometry-based methods were used to detect the global EV proteome and phosphoproteome. Differentially expressed features, enriched pathways, and enriched tissue-specific protein sets were identified. Multidimensional scaling of all EV proteomic and phosphoproteomic data and unsupervised clustering of differentially expressed (absolute fold change ≥ 2, P < 0.05, FDR < 0.05) EV proteins and phosphoproteins successfully distinguished the 4 study groups with close to 100% accuracy. Importantly, we detected enriched pathway networks that suggest the potential therapeutic utility of PKC inhibitors such as bisindolylmaleimide IX, sotrastaurin, and enzastaumn, and inhibitors of ROCK1 such the isoquinoline derivative Fasudil. In conclusion, we characterized the proteomic landscape of the interaction between type 2 diabetes and COVID-and defined disease-specific EV proteomic signatures that provide insight into the disease pathobiology and druggable targets with potential clinical utility.