SARS-CoV-2 Infected Pediatric Cerebral Cortical Neurons: Transcriptomic Analysis and Potential Role of Toll-like Receptors in Pathogenesis

Int J Mol Sci. 2021 Jul 28;22(15):8059. doi: 10.3390/ijms22158059.

Abstract

Different mechanisms were proposed as responsible for COVID-19 neurological symptoms but a clear one has not been established yet. In this work we aimed to study SARS-CoV-2 capacity to infect pediatric human cortical neuronal HCN-2 cells, studying the changes in the transcriptomic profile by next generation sequencing. SARS-CoV-2 was able to replicate in HCN-2 cells, that did not express ACE2, confirmed also with Western blot, and TMPRSS2. Looking for pattern recognition receptor expression, we found the deregulation of scavenger receptors, such as SR-B1, and the downregulation of genes encoding for Nod-like receptors. On the other hand, TLR1, TLR4 and TLR6 encoding for Toll-like receptors (TLRs) were upregulated. We also found the upregulation of genes encoding for ERK, JNK, NF-κB and Caspase 8 in our transcriptomic analysis. Regarding the expression of known receptors for viral RNA, only RIG-1 showed an increased expression; downstream RIG-1, the genes encoding for TRAF3, IKKε and IRF3 were downregulated. We also found the upregulation of genes encoding for chemokines and accordingly we found an increase in cytokine/chemokine levels in the medium. According to our results, it is possible to speculate that additionally to ACE2 and TMPRSS2, also other receptors may interact with SARS-CoV-2 proteins and mediate its entry or pathogenesis in pediatric cortical neurons infected with SARS-CoV-2. In particular, TLRs signaling could be crucial for the neurological involvement related to SARS-CoV-2 infection.

Keywords: SARS-CoV-2; Toll-like receptors; children; human neuronal cells; nervous system; transcriptomic analysis.

MeSH terms

  • COVID-19 / genetics
  • COVID-19 / immunology
  • COVID-19 / metabolism*
  • Cerebral Cortex / metabolism*
  • Child
  • Cytokines / metabolism
  • Gene Expression Profiling
  • Gene Expression Regulation
  • Humans
  • Neurons / immunology
  • Neurons / virology*
  • SARS-CoV-2 / immunology
  • SARS-CoV-2 / metabolism
  • SARS-CoV-2 / pathogenicity*
  • Signal Transduction / genetics
  • Toll-Like Receptors / genetics
  • Toll-Like Receptors / metabolism*
  • Virus Replication

Substances

  • Cytokines
  • Toll-Like Receptors