A journey upstream: Fluctuating platelet-specific genes in cell-free plasma as proof-of-concept for using ribonucleic acid sequencing to improve understanding of postinjury platelet biology

Lucy Zumwinkle Kornblith, Cedric M.V. Bainton, Alexander T. Fields, Zachary A. Matthay, Nina T. Magid, Brenda Nunez-Garcia, Arun Prakash, Philip A. Kurien, Rachael A. Callcut, Mitchell J. Cohen, Roland J. Bainton

Research output: Contribution to journalArticlepeer-review

Abstract

BACKGROUND The mechanisms of aberrant circulating platelet behavior following injury remain unclear. Platelets retain megakaryocyte immature ribonucleic acid (RNA) splicing and protein synthesis machinery to alter their functions based on physiologic signals. We sought to identify fluctuating platelet-specific RNA transcripts in cell-free plasma (CFP) from traumatic brain injury (TBI) patients as proof-of-concept for using RNA sequencing to improve our understanding of postinjury platelet behavior. We hypothesized that we could identify differential expression of activated platelet-specific spliced RNA transcripts from CFP of patients with isolated severe fatal TBI (fTBI) compared with minimally injured trauma controls (t-controls), filtered by healthy control (h-control) data sets. METHODS High-read depth RNA sequencing was applied to CFP from 10 patients with fTBI (Abbreviated Injury Scale [AIS] for head ≥3, AIS for all other categories <3, and expired) and five t-controls (Injury Severity Score ≤1, and survived). A publicly available CFP RNA sequencing data set from 23 h-controls was used to determine the relative steady state of splice-form RNA transcripts discoverable in CFP. Activated platelet-specific spliced RNA transcripts were derived from studies of ex vivo platelet activation and identified by splice junction presence greater than 1.5-fold or less than 0.67-fold ex vivo nonactivated platelet-specific RNA transcripts. RESULTS Forty-two differentially spliced activated platelet-specific RNA transcripts in 34 genes were altered in CFP from fTBI patients (both upregulated and downregulated). CONCLUSION We have discovered differentially expressed activated platelet-specific spliced RNA transcripts present in CFP from isolated severe fTBI patients that are upregulated or downregulated compared with minimally injured trauma controls. This proof-of-concept suggests that a pool of immature platelet RNAs undergo splicing events after injury for presumed modulation of platelet protein products involved in platelet function. This validates our exploration of injury-induced platelet RNA transcript modulation as an upstream "liquid biopsy" to identify novel postinjury platelet biology and treatment targets for aberrant platelet behavior. LEVEL OF EVIDENCE Diagnostic tests, level V.

Original languageEnglish (US)
Pages (from-to)742-751
Number of pages10
JournalJournal of Trauma and Acute Care Surgery
Volume88
Issue number6
DOIs
StatePublished - Jun 1 2020
Externally publishedYes

Keywords

  • cell-free plasma
  • dynamically regulated RNA splicing
  • next generation sequencing
  • Platelet activation
  • prespliced RNA complexes
  • ribonucleic acid sequencing

ASJC Scopus subject areas

  • Surgery
  • Critical Care and Intensive Care Medicine

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