Evaluating the feasibility of platelet-derived gene expression profiling in dogs with sepsis.

Abstract

Objective: To assess the feasibility of gene expression profiling of platelets in dogs with sepsis.

Methods: This was a prospective observational feasibility study conducted at a university veterinary teaching hospital. All study dogs had CBCs and serum biochemistry profiles, and blood samples were collected for platelet isolation. Gene expression profiling of isolated platelets was performed using a commercial multiplex assay based on direct hybridization and detection of single RNA molecules bar coded with fluorescent probes. Quality control, normalization, and data visualization were performed using standardized workflows. Raw counts were exported, and differential expression between groups was assessed.

Results: 6 client-owned dogs with sepsis and 6 healthy dogs were enrolled from March through August 2023. Platelet isolation and transcript profiling were successful for all dogs. The septic dog group included 3 males and 3 females with peritonitis (n = 3), pyometra (n = 2), and pyothorax. Principal component analysis did not fully segregate septic from healthy dogs, but disease status accounted for 59% of the variance. Upregulated genes contributing substantially to principal component analysis variance included those for S100 calcium-binding proteins (S100s) and cytokines. Significantly upregulated genes based on log2 fold change included S100A8, S100A12, cathelicidin, lactoferrin, and innate immune response signaling proteins.

Conclusions: Sample processing and data analysis workflows are scalable to larger, confirmatory studies to explore potential sepsis biomarkers.

Clinical relevance: Platelet gene expression profiles might offer insights into sepsis pathophysiology and potential diagnostic and prognostic biomarkers. Follow-up studies are warranted to characterize platelet expression profiles in larger populations of dogs with sepsis and noninfectious critical illnesses.