Abstract 147: Novel Inflammatory and Pro-Thrombotic mRNA Pathways Associated With Worse 3-month Outcome After Intracerebral Hemorrhage

Abstract

Background: Intracerebral hemorrhage (ICH) has the highest disability among stroke survivors. The Recovery and Outcomes from Stroke (ROSE) study (U01NS100417) utilized RNA sequencing (RNAseq) to identify biomarkers associated with poor outcome at 3 months. Methods: Supratentorial, non-moribund, ≥ 18 years of age, spontaneous ICH patients were prospectively recruited in a multi-center study. Whole blood was obtained at baseline (<15 days; RNA PAXgene tubes, Qiagen). 3-month outcomes were determined by modified Rankin Scale (mRS) 0-3 (good) vs 4-6 (poor). RNAseq libraries were constructed using the Illumina TruSeq Stranded Total RNA kit with Ribo-Zero, sequenced on an llumina NovaSeq 6000 platform using 1 x100 bp single end reads to generate 25-30 million reads at >90% >Q30 reads. Differential expression was tested using DESeq2 (negative binomial), adjusting for sequencing batch, age, sex, race, ICH volume, intraventricular hemorrhage, presenting Glasgow Coma Scale, and lobar vs non-lobar location. Pathway analyses and protein-protein interactions were computed on a set of genes (P < 0.0001) using STRING, Cytoscape, MCODE, and Reactome. Results: We analyzed baseline blood samples from 278 patients (N = 156 vs 122; good vs poor outcomes at 3 months). Multiple pathways were associated with poor outcome, including those involved in clot formation and cell-cell/cell-matrix interactions (not shown). The statistically most significant pathway was observed for transcripts involved in neutrophil activation and degranulation. Conclusion: Variation in mRNA expression pathways associated with inflammation, coagulation and cellular interactions were associated with poor outcome after controlling for severity measures.