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Cited by 13Neonatal and fetal brain pathologyNeuroinflammation and Neurodegeneration MechanismsNeonatal and Maternal Infections

Serial blood cytokine and chemokine mRNA and microRNA over 48 h are insult specific in a piglet model of inflammation-sensitized hypoxia–ischaemia

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Lingam, Ingran, Avdic-Belltheus, Adnan, Meehan, Christopher, Martinello, Kathryn, Ragab, Sara, Peebles, Donald, Barkhuizen, Melinda, Tann, Cally J. +9 more·2021·Pediatric Research
DOI 10.1038/s41390-020-0986-3
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Abstract

Background Exposure to inflammation exacerbates injury in neonatal encephalopathy (NE). We hypothesized that brain biomarker mRNA, cytokine mRNA and microRNA differentiate inflammation (E. coli LPS), hypoxia (Hypoxia), and inflammation-sensitized hypoxia (LPS+Hypoxia) in an NE piglet model. Methods Sixteen piglets were randomized: (i) LPS 2 μg/kg bolus; 1 μg/kg infusion (LPS; n = 5), (ii) Saline with hypoxia (Hypoxia; n = 6), (iii) LPS commencing 4 h pre-hypoxia (LPS+Hypoxia; n = 5). Total RNA was acquired at baseline, 4 h after LPS and 1, 3, 6, 12, 24, 48 h post-insult (animals euthanized at 48 h). Quantitative PCR was performed for cytokines (IL1A, IL6, CXCL8, IL10, TNFA) and brain biomarkers (ENO2, UCHL1, S100B, GFAP, CRP, BDNF, MAPT). MicroRNA was detected using GeneChip (Affymetrix) microarrays. Fold changes from baseline were compared between groups and correlated with cell death (TUNEL) at 48 h. Results Within 6 h post-insult, we observed increased IL1A, CXCL8, CCL2 and ENO2 mRNA in LPS+Hypoxia and LPS compared to Hypoxia. IL10 mRNA differentiated all groups. Four microRNAs differentiated LPS+Hypoxia and Hypoxia: hsa-miR-23a, 27a, 31-5p, 193-5p. Cell death correlated with TNFA (R = 0.69; p < 0.01) at 1-3 h and ENO2 (R = -0.69; p = 0.01) at 48 h. Conclusions mRNA and miRNA differentiated hypoxia from inflammation-sensitized hypoxia within 6 h in a piglet model. This information may inform human studies to enable triage for tailored neuroprotection in NE. Impact Early stratification of infants with neonatal encephalopathy is key to providing tailored neuroprotection. IL1A, CXCL8, IL10, CCL2 and NSE mRNA are promising biomarkers of inflammation-sensitized hypoxia. IL10 mRNA levels differentiated all three pathological states; fold changes from baseline was the highest in LPS+Hypoxia animals, followed by LPS and Hypoxia at 6 h. miR-23, -27, -31-5p and -193-5p were significantly upregulated within 6 h of a hypoxia insult. Functional analysis highlighted the diverse roles of miRNA in cellular processes.

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  • Dynamic changes in miR-23 expression patterns at different time points enable it to serve as a biomarker to distinguish different states of inflammation, hypoxia, and inflammation-sensitive hypoxia.

    “The dynamic changes in expression patterns at different time points enable it to serve as a biomarker to distinguish and differentiate different states of inflammation, hypoxia, and inflammation sensitive hypoxia [41].”

    From Recent advances of miR-23 in human diseases and growth development (2025) · cites this paper as [41]

    The ‘Expression of microRNA’ section (page unavailable) reports that hsa-miR-23a-5p differed at 1, 3 and 6 hours: levels were higher in both LPS and LPS-plus-hypoxia groups than in the hypoxia group. The ‘MiRNA Biomarkers’ section calls it a promising early differentiating biomarker, but the reported results do not show that miR-23 distinguishes inflammation alone from inflammation-sensitized hypoxia. (LLM, conf=0.87)

    Experimental — accuracy not yet measured

  • In a pig model of encephalopathy, miR-23 was significantly upregulated within 6 h after hypoxic injury.

    “Additionally, in an established pig model of encephalopathy, it was observed that miR-23 was significantly upregulated within 6 h after hypoxic injury.”

    From Recent advances of miR-23 in human diseases and growth development (2025) · cites this paper as [41]

    The Expression of microRNA section (page unavailable) reports that miR-23a-5p was significantly higher at 1, 3, and 6 hours in piglets given LPS plus hypoxia than in those given hypoxia alone. The claim omits the LPS sensitization and the comparison group, making the increase sound like a general response to hypoxic injury; the section reports that miRNAs significantly changed from baseline in the hypoxia-only group were all downregulated. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

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Research works printed but not linked: 39

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Authors
Lingam, I., Avdic-Belltheus, A., Meehan, C., Martinello, K., Ragab, S., Peebles, D., Barkhuizen, M., Tann, C. J., Tachtsidis, I., Wolfs, T. G. A. M., Hagberg, H., Klein, N., Fleiss, B., Gressens, P., Golay, X., Kramer, B. W., & Robertson, N. J.
Affiliations
  • University College London (GB)
  • Maastricht University (NL)
  • London School of Hygiene & Tropical Medicine (GB)
  • University of Gothenburg (SE)
  • King's College London (GB)
  • UCL Queen Square Institute of Neurology (GB)
Corresponding author
Robertson, Nicola J.
ORCID
Venue
Pediatric Research
Publisher
Springer Science and Business Media LLC
Published
2020-06-10
Volume
89
Issue
3
Pages
464-475
ISSN
  • 0031-3998 (ISSN-L)
  • 1530-0447
DOI
10.1038/s41390-020-0986-3
OpenAlex
W3033311404
Cited by
13
References
53
Citations by year
2025: 3 · 2024: 4 · 2022: 1 · 2021: 4 · 2020: 1
FWCI
1.44
Type
journal article
Language
en
Open access
Licence
Locations
  • Pediatric Research (publishedVersion)
  • PubMed (publishedVersion)
  • Research Publications (Maastricht University) (acceptedVersion, other-oa)
  • LSHTM Research Online (London School of Hygiene and Tropical Medicine) (acceptedVersion)
  • RMIT Research Repository (RMIT University Library) (submittedVersion)
  • UCL Discovery (University College London) (submittedVersion)
  • Figshare (submittedVersion, cc-by)
  • Research Publications (Maastricht University) (submittedVersion)
Funders
  • National Institute for Health and Care Research
  • University College London
  • Agence Nationale de la Recherche (ANR-11-INBS-0011)
  • Medical Research Council (MR/M006743/1)
  • National Heart, Lung, and Blood Institute (T32HL105349)
  • National Institute of Diabetes and Digestive and Kidney Diseases (P30DK056336, P30DK079626)
  • National Institute on Minority Health and Health Disparities (U54MD000502)
Topics
  • Health Sciences › Medicine › Pediatrics, Perinatology and Child Health › Neonatal and fetal brain pathology
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
  • Health Sciences › Medicine › Public Health, Environmental and Occupational Health › Neonatal and Maternal Infections
Keywords
neonatal encephalopathy, lipopolysaccharide, cytokine mRNA, microRNA, brain biomarkers, neuron-specific enolase, UCHL1, S100B, glial fibrillary acidic protein, brain-derived neurotrophic factor, MAPT, TNF-α, interleukin-1 alpha, interleukin-8, cell death, neuroprotection, piglet model
MeSH terms
Animals, Animals, Newborn, Brain, Disease Models, Animal, Gene Expression Regulation, Humans, Inflammation, Lipopolysaccharides, Male, Nerve Tissue Proteins, Phosphopyruvate Hydratase, Random Allocation, RNA, Messenger, Swine, Time Factors, Tumor Necrosis Factor-alpha, Biomarkers, Cytokines, Chemokines, Endotoxemia, Hypoxia-Ischemia, Brain, MicroRNAs, Tissue Array Analysis, Gene Ontology, Sepsis-Associated Encephalopathy

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