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Gene expression response to simulated gravity and hypergravity in human T cells

ABSTRACT: We investigated differentially regulated and stably expressed genes in human Jurkat T lymphocytic cells in 5min simulated microgravity and hypergravity and compared expression profiles to identify gravity-regulated and unaffected genes as well as adaptation processes.

ORGANISM(S): Homo sapiens

PROVIDER: GSE101101 | GEO | 2018/03/08

REPOSITORIES: GEO

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Project description:Purpose: Microgravity is one of crucial factors which affects astronauts’ health in space. The goals of this study are to compare SMG-pretreated macrophages infected with EPEC by RNA-sequencing to investigate the immunity-related differential expression genes and signaling pathway and to make for prevention and therapy of infectious diseases in space. Methods: 3-day SMG-pretreatment macrophages were infected with EPEC (a kind of enteropathogen) and macrophages infected with EPEC under normal gravity as control and then we used RNA sequencing on Illumina platform. The sequence reads that passed quality filters were analyzed at the transcript isoform level by Volcano Plot, KEEG database and InnateDB database. qRT–PCR validation was performed using SYBR Green assays. Results: After RNA sequencing, we got 1272 differential expression genes (DEGs) of which 542 were up-regulated, 720 were down-regulated DEGs. KEGG pathway enrichment scatter plot showed that cytokine-cytokine receptor interaction and MAPK signaling pathway were optimal-related to infection immunity under simulated microgravity. Because of our study involved in infection immunity, we chose a database on immunity called InnateDB to screen the DEGs related to immunity. We found 14 DEGs in cytokine-cytokine receptor interaction, of which 7 were up-regulated, and 7 were down-regulated and 36 DEGs in MAPK signaling pathway, of which 11 were up-regulated, and 25 were down-regulated. In cytokine-cytokine receptor interaction signaling pathway, except for Il 33 (P>0. 05), the results of mRNA expression of DEGs in macrophages infected with EPEC were consistent with the results of RNA sequencing. In MAPK signaling pathway, the mRNA expression of Il 1b, Cd14, Hspa1a, Pdgfrb, Il 1a, Fgf7, Igf1r and Pak1 was not consistent with the RNA sequencing. Compared with normal gravity, simulated microgravity didn’t affect the mRNA expression of Il 1b, Cd14, Hspa1a and Pdgfrb (P>0. 05). The Il 1a and Fgf7 down regulated and Igf1r and Pak1 up regulated under simulated microgravity at the level of mRNA with a comparison to normal gravity, and the results of the rest of DEGs were consistent with the results of RNA sequencing. Conclusions: This study is the first high-throughput analysis on simulated microgravity of infection immunity. The comparative analysis demonstrates that cytokine-cytokine receptor interaction and MAPK signaling pathway participants in the innate immune response.

Project description:Genome-wide transcriptional profiling shows that reducing gravity levels in the International Space Station (ISS) causes important alterations in Drosophila gene expression. However, simulation experiments on ground, without space constraints, show weaker effects than space environment. A global and integrative analysis using the M-bM-^@M-^\gene expression dynamics inspectorM-bM-^@M-^] (GEDI) self-organizing maps, reveals a subtle response of the transcriptome using different populations and microgravity and hypergravity simulation devices. These results suggest that, in addition to behavioural responses that can be detected also at the gene expression level, the transcriptome is finely tuned to normal gravity. The alteration of this constant parameter on Earth can have effects on gene expression that depends both on the environmental conditions and the ground based facility used to compensate the gravity vector. Alternative and commons effects of mechanical facilities, like the Random Positioning Machine and a centrifuge, and strong magnetic field ones, like a cryogenically cooled superconductive magnet, are discussed. We compare the effects over the gene expression profile of different gender/age Drosophila imagoes in 3-4 days-long experiments under altered gravity conditions into three GBF ("Ground Based Facilities" for micro/hyper- gravity simulation) using whole genome microarray platforms. Descriptions of different GBFs ("treatments"): LDC means "Large Diameter Centrifuge". Samples can be placed under three conditions: inside LDC (at certain g level), at the LDC rotational control and at external 1g control (outside the LDC). RPM means "Random Positioning Machine". Samples can be placed under two conditions: inside RPM (at nearly 0g, Microgravity level) and at external 1g control (outside the RPM). At the magnet, means INSIDE the Magnetic levitator (another GBF). Samples can be placed under four conditions: inside Magnet 0g* (at microgravity with magnetic field), inside Magnet at 1g* (internal control with magnetic field) or inside the magnet 2g* (at hypergravity with magnetic field) and at external 1g control (outside the magnet)

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Gene expression response to simulated gravity and hypergravity in human T cells

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