Impact of Congenital Cytomegalovirus Infection on Transcriptomes From Archived Dried Blood Spots in Relation to Long-Term Clinical Outcome

PLoS One. 2018 Jul 19;13(7):e0200652. doi: 10.1371/journal.pone.0200652. eCollection 2018.

Abstract

Congenital Cytomegalovirus infection (cCMV) is the leading infection in determining permanent long-term impairments (LTI), and its pathogenesis is largely unknown due to the complex interplay between viral, maternal, placental, and child factors. The cellular activity, considered to be the result of the response to exogenous and endogenous factors, is captured by the determination of gene expression profiles. In this study, we determined whole blood transcriptomes in relation to cCMV, CMV viral load and LTI development at 6 years of age by using RNA isolated from neonatal dried blood spots (DBS) stored at room temperature for 8 years. As DBS were assumed to mainly reflect the neonatal immune system, particular attention was given to the immune pathways using the global test. Additionally, differential expression of individual genes was performed using the voom/limma function packages. We demonstrated feasibility of RNA sequencing from archived neonatal DBS of children with cCMV, and non-infected controls, in relation to LTI and CMV viral load. Despite the lack of statistical power to detect individual genes differences, pathway analysis suggested the involvement of innate immune response with higher CMV viral loads, and of anti-inflammatory markers in infected children that did not develop LTI. Finally, the T cell exhaustion observed in infected neonates, in particular with higher viral load, did not correlate with LTI, therefore other mechanisms are likely to be involved in the long-term immune dysfunction. Despite these data demonstrate limitation in determining prognostic markers for LTI by means of transcriptome analysis, this exploratory study represents a first step in unraveling the pathogenesis of cCMV, and the aforementioned pathways certainly merit further evaluation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blood Preservation / methods*
  • Child
  • Child, Preschool
  • Cognitive Dysfunction / diagnosis
  • Cytomegalovirus / physiology
  • Cytomegalovirus Infections / blood
  • Cytomegalovirus Infections / genetics*
  • Cytomegalovirus Infections / virology
  • Dried Blood Spot Testing / methods*
  • Female
  • Humans
  • Infant, Newborn
  • Male
  • Motor Neuron Disease / diagnosis
  • Time Factors
  • Transcriptome*
  • Viral Load

Grant support

This work was supported by European Union Seventh Framework Programme FP7/2012–2016 under grant agreement number 316655 (VACTRAIN). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. HL is a founder and the chairman of Alacris Theranostics GmbH, Berlin, Germany. Alacris Theranostics GmbH provided support in the form of salary for author HL, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific role of this author is articulated in the ‘author contributions’ section.