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| Status |
Public on Oct 31, 2018 |
| Title |
Genome-wide lung transcriptional profiling of mouse models for pneumonia given human adipose-derived mesenchymal stem cells treatment or Prekallikrein |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
Adult mesenchymal stem cells exert immunomodulatory effects that might improve the host response during sepsis. Knowledge on the effect of adipocyte-derived mesenchymal stem cells (ASCs) in sepsis is limited. Klebsiella (K.) pneumoniae is a common cause of gram-negative pneumonia and sepsis. The aim of this study was to determine the effect of human ASCs on the host response during pneumosepsis in mice. For this mice were infected with K. pneumoniae via the airways to induce a gradually evolving infection in the lung cumulating in pneumosepsis. One or 6 hours (h) after infection, mice were infused intravenously with ASCs or vehicle, and euthanized after 16h or 48h respectively. The effects of freshly cultured and cryopreserved ASCs were compared, the latter formulation being more clinically relevant. Intravenously administered ASCs were visualized in lung tissue by immunostaining at 1 and 3h, but not at 15h after infusion. While early after infection ASCs did not or only modestly influence bacterial loads, they reduced bacterial burdens in lungs and distant organs at 48h. ASCs reduced the lung levels of pro-inflammatory cytokines and attenuated lung pathology, but did not influence distant organ injury. ASCs strongly modified the lung transcriptome in both uninfected mice and mice with pneumosepsis, especially affecting. Cryopreserved and cultured ASCs induced largely similar effects. These data indicate that human ASCs induce profound immune modulatory effects in the lungs, resulting in reduced bacterial burdens and lung inflammation during pneumosepsis caused by a common human pathogen, suggesting that ASCs may be an adjunctive therapeutic in this condition.
Experimental groups were defined as follows: (1) plac_cryo, placebo treated and K.pneumoniae infected control mice for cryopreserved MSCs; (2) plac_cult, placebo treated and K.pneumoniae infected control mice for cultured MSCs; (3) cryo, K.pneumoniae infected mice treated with cryopreserved MSCs; (4) cult, K.pneumoniae infected mice treated with cultured MSCs; (5) n_cult, non-infected control mice for cultured MSCs; (6) n_cryo, non-infected control mice for cryopreserved MSCs; (7) n, normal mouse lung baseline.
In a separate experiment, mice were treated with a selective prekallikrein (PKK) directed antisense oligonucleotide (ASO) or a scrambled control ASO for 3 weeks prior to infection. Host response readouts were determined at 12 or 36 hours postinfection, including genome-wide messenger RNA profiling of lungs, or mice were followed for survival
Pre-processing and quality control of the scans were performed by using the oligo method (version 1.44) and probes were annotated using the platform design info for Affymetrix Clariom_S_Mouse_HT available through Bioconductor. Array data were background corrected by Robust Multi-array Average (RMA) and quantiles-normalized. Microarray quality control was performed by means of the array quality metrics method (version 3.36.0). The occurrence of non-experimental chip effects was evaluated by the surrogate variable analysis method (version 3.28.0) and corrected using the combat method.
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| Overall design |
Pneumonia was induced by intranasal inoculation with 10^4 colony forming units (CFU) K. pneumoniae serotype 2 (ATCC 43816, Rockville, MD). In all human-derived mesenchymal stem cell experiments mice were intravenously (i.v.) infused with 1 x 10^6 adipose derived stem cells (ASCs) at 1 or 6 hours after infection. In a separate experiment, mice were treated with a selective prekallikrein (PKK) directed antisense oligonucleotide (ASO) or a scrambled control ASO for 3 weeks prior to infection. We performed genome-wide scans of pulmonary gene expression in uninfected and infected mice treated with ASCs. For this analysis we selected the 16 hour time point to avoid confounding effects by differences in bacterial burden observed at later time points. RNA was isolated from lung homogenate using the Nucleospin RNA isolation kit (Macherey-Nagel, Düren, Germany) as described by the manufacturer. RNA samples with integrity number (RIN) > 6 (Agilent Bioanalyzer) were included for microarrays.
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| Contributor(s) |
Scicluna BP, van der Poll T |
| Citation(s) |
31033196, 31547876, 31595971 |
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| Submission date |
Oct 30, 2018 |
| Last update date |
Feb 03, 2020 |
| Contact name |
Brendon Scicluna |
| E-mail(s) |
brendon.scicluna@um.edu.mt
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| Organization name |
University of Malta
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| Street address |
Msida campus
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| City |
Msida |
| ZIP/Postal code |
MSD2020 |
| Country |
Malta |
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| Platforms (1) |
| GPL24242 |
[Clariom_S_Mouse_HT] Affymetrix Clariom S Assay HT, Mouse (Includes Pico Assay) |
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| Samples (47)
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| Relations |
| BioProject |
PRJNA501867 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE121970_RAW.tar |
51.8 Mb |
(http)(custom) |
TAR (of CEL) |
| Processed data included within Sample table |
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