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Transcriptomic Profiling and Pathway Analysis of Cultured Human Lung Microvascular Endothelial Cells following Ionizing Radiation Exposure

ABSTRACT: Using RNA-seq we report changes in gene expression, over a 24 hour time course, in human lung microvascular endothelial cells subjected to 10 Gy X-irradiation.

ORGANISM(S): Homo sapiens

PROVIDER: GSE179810 | GEO | 2022/07/01

REPOSITORIES: GEO

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Transcriptomic profiling and pathway analysis of cultured human lung microvascular endothelial cells following ionizing radiation exposure.

Project description:The vascular system is sensitive to radiation injury, and vascular damage is believed to play a key role in delayed tissue injury such as pulmonary fibrosis. However, the response of endothelial cells to radiation is not completely understood. We examined the response of primary human lung microvascular endothelial cells (HLMVEC) to 10 Gy (1.15 Gy/min) X-irradiation. HLMVEC underwent senescence (80-85%) with no significant necrosis or apoptosis. Targeted RT-qPCR showed increased expression of genes CDKN1A and MDM2 (10-120 min). Western blotting showed upregulation of p2/waf1, MDM2, ATM, and Akt phosphorylation (15 min-72 h). Low levels of apoptosis at 24-72 h were identified using nuclear morphology. To identify novel pathway regulation, RNA-seq was performed on mRNA using time points from 2 to 24 h post-irradiation. Gene ontology and pathway analysis revealed increased cell cycle inhibition, DNA damage response, pro- and anti- apoptosis, and pro-senescence gene expression. Based on published literature on inflammation and endothelial-to-mesenchymal transition (EndMT) pathway genes, we identified increased expression of pro-inflammatory genes and EndMT-associated genes by 24 h. Together our data reveal a time course of integrated gene expression and protein activation leading from early DNA damage response and cell cycle arrest to senescence, pro-inflammatory gene expression, and endothelial-to-mesenchymal transition.

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Genome-wide DNA damage response following ionizing radiation

Project description:H2AX phosphorylation at Ser139 (gH2AX) is an early key event of the DNA damage response (DDR) following DNA double strand breaks (DSB) induction. Although gH2AX distribution has been extensively used as a damage marker, genome-wide investigation of the subsequent DNA repair has been poorly addressed. Here we present ChIP-Seq-based distributions of Histone H3, H2AX and gH2AX under physiological conditions in HeLa cells. Additionally the same histones were studied after a single exposure to 10 Gy X-ray at 0.5, 3 and 24 hours post irradiation

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Ionizing radiation exposure of developing mouse brain and cultured neurons

Project description:The mammalian brain is especially sensitive to ionizing radiation during development, as shown by the increased occurrence of mental retardation and small head size in children who were in utero exposed to ionizing radiation after the atomic bombings of Hiroshima and Nagasaki. These effects of prenatal irradiation can be mimicked by irradiation of mouse embryos during the organogenesis period. In order to better understand the early effects of ionizing radiation on the embryonic brain and immature neurons, we performed a microarray analysis on brains from mice irradiated with different doses at E11 and E14, as well as primary cortical neuron cultures after 14 h in vitro. RNA was extracted at either 2 h (brains) or 6 h (neurons) post-irradiation.

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Assessment of host responses to ionizing radiation

Project description:The aim of this study was to analyze the host responses to ionizing radiation by nuclear factor-κB (NF-κB) bioluminescence imaging-guided transcriptomic tool. Transgenic mice, carrying the NF-κB-driven luciferase gene, were exposed to a single dose of 8.5 Gy total-body irradiation. In vivo imaging showed that a maximal NF-κB-dependent bioluminescent intensity was observed at 3 h after irradiation and ex vivo imaging showed that liver, intestine, and brain displayed strong NF-κB activations. Microarray analysis of these organs showed that irradiation altered gene expression signatures in an organ-specific manner. Pathway analysis showed that pathways associated with metabolism and immune system were altered primarily in liver and intestine. the upregulation of fatty acid binding protein 4, serum amyloid A2, and serum amyloid A3, which are participated in both inflammation and lipid metabolism, suggesting that irradiation might affect the cross pathways of metabolism and inflammation. Moreover, The upregulation of chemokine (CC-motif) ligand 5, chemokine (CC-motif) ligand 20, and Jagged 1 genes suggested that these genes might contribute to the radiation enteropathy. Male transgenic mice (6 to 8 weeks old) were exposed to a single dose of whole-body X-rays generated at 6 MV (Clinac® 21EX medical linear accelerator, Varian, Palo Alto, CA, USA) and at a dose rate of 4 Gy/min. Mice were imaged at 0 h, 1 h, 3 h, 9 h, 24 h, 48 h, or 72 h, or on 7 d or 14 d after irradiation with 8.5 Gy. RNAs were extracted at 3 h after irradiation.

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X-ray causes mRNA transcripts change to enhance Orai2-meadated Ca2+ influx in rat brain microvascular endothelial cells

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Transcriptomic Profiling and Pathway Analysis of Cultured Human Lung Microvascular Endothelial Cells following Ionizing Radiation Exposure

Project description:Transcriptomic Profiling and Pathway Analysis of Cultured Human Lung Microvascular Endothelial Cells following Ionizing Radiation Exposure

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Ionizing radiation induced hair and skin damage

Project description:Skin samples from back region and footpads were analyzed in 8-10 week old mice. For back skin, the hair was plucked in order to induce active growing phase(anagen). After 9 days, the mice were irradiated for 5Gy IR. Krt17 knock out, p53 knock out, and wild type mice were compared. Samples were taken at time 0 (control), 6h, and 24h post-irradiation. For footpad skin, the mice were irradiated for 40 Gy IR. Krt17 knock out and wild type mice were compared. Samples were taken at time 0 (control), 3 d, 6 d, 9 d post-irradiation.

2020-12-22 | GSE163653 | GEO

Transcription profiling of human lymphoblastoid cells following 3 Gy and 10 Gy of ionizing radiation exposure to identify transcriptional response of lymphoblastoid cells to ionizing radiation

Project description:Time series for gene expression changes following 3 Gy and 10 Gy of ionizing radiation exposure.

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RNA-seq of cultured human kidney peritubular microvascular endothelial cells following exposure to cyclosporine A

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