Find research datasets worth reusing

Comparative gene expression analysis in the Arabidopsis thaliana root apex using RNA-seq and microarray transcriptome profiles

The root apex is an important section of the plant root involved in environmental sensing and cellular development. Analyzing the gene profile of root apex in diverse environments is important and challenging especially when the samples are limiting and precious such as in spaceflight. The feasibility of using tiny root sections for transcriptome analysis was examined in this study. To understand the gene expression profiles of the root apex Arabidopsis thaliana Col-0 roots were sectioned into Zone-I (0.5 mm root cap and meristematic zone) and Zone-II (1.5 mm transition elongation and growth terminating zone). Gene expression was analyzed using microarray and RNA seq. Both the techniques arrays and RNA-Seq identified 4180 common genes as differentially expressed (with > two-fold changes) between the zones. In addition 771 unique genes and 19 novel TARs were identified by RNA-Seq as differentially expressed which were not detected in the arrays. Single root tip zones can be used for full transcriptome analysis; further the root apex zones are functionally very distinct from each other. RNA-Seq provided novel information about the transcripts compared to the arrays. These data will help optimize transcriptome techniques for dealing with small rare samples.

Global gene expression profiles of cardiac progenitors differentiated from human pluripotent stem cells in 3D culture under simulated microgravity

Efficient generation of cardiomyocytes from human pluripotent stem cells is critical for their regenerative applications. Microgravity and 3D culture can profoundly modulate cell proliferation and survival. Here we engineered microscale progenitor cardiac spheres from human pluripotent stem cells and exposed the spheres to simulated microgravity using a random positioning machine for 3 days during their differentiation to cardiomyocytes. Methods: RNA-seq libraries were prepared using the Illumina TruSeq RNA kit and the TrueSeq method was employed for mRNA enrichment. The libraries were quantified and samples were multiplexed in each lane of the flowcell. Cluster generation was performed and then sequenced on the Illumina HiSeq1000 system. Reads were mapped on the Human Genome Reference and normalized expression table was generated. Results: Among differentially expressed genes 53 of them were up-regulated and 75 were down-regulated. Conclusions: Data demonstrate increased expression of genes associated with growth development and pro-survival in cardiac progenitors cultured under simulated microgravity compared with those cultured under standard gravity. RNA-sequencing analysis was performed to compare global gene expression profiles of cells at differentiation day 8 under simulated microgravity vs. standard gravity.

Effects of a Closed Space Environment on Gene Expression in Hair Follicles of Astronauts in the International Space Station

In recent times long-term stay has become a common occurrence in the International Space Station (ISS). However adaptation to the space environment can sometimes pose physiological problems to the astronauts after their return. Therefore it is important to develop healthcare technologies for astronauts. In this study hair an easy-to-obtain sample was identified as the candidate. In order to investigate the genetic changes in human hair during space flight the hair follicles of 10 astronauts were analyzed by DNA microarray and real time q-PCR analyses. Space environment induced gene expression of hair follicles of astronaut was measured 6 differnent times included 2 in flight on orbit. Ten independent experiments were performed on differing astronauts. and the sampling day was differed for each astronaut because of their schedules.

Gene expression data from 4T1 irradiated tumors treated with TGFbeta blockade

Accumulating data support the concept that ionizing radiation therapy (RT) has the potential to convert the tumor into an in situ individualized vaccine; however this potential is rarely realized by RT alone. Transforming growth factor xce xb2 (TGF xce xb2) is an immunosuppressive cytokine that is activated by RT and inhibits the antigen-presenting function of dendritic cells and the differentiation of effector CD8+ T cells. Here we tested the hypothesis that TGF xce xb2 hinders the ability of RT to promote anti-tumor immunity. Development of tumor-specific immunity was examined in a pre-clinical model of metastatic breast cancer. Mice bearing established 4T1 mouse mammary carcinoma treated with pan-isoform specific TGF xce xb2 neutralizing antibody 1D11 showed significantly improved control of the irradiated tumor and non-irradiated metastases but no effect in the absence of RT. Notably whole tumor transcriptional analysis demonstrated the selective upregulation of genes associated with immune-mediated rejection only in tumors of mice treated with RT+TGF xce xb2 blockade. Mice treated with RT+TGF xce xb2 blockade exhibited cross-priming of CD8+ T cells producing IFN xce xb3 in response to three tumor-specific antigens in tumor-draining lymph nodes which was not evident for single modality treatment. Analysis of the immune infiltrate in mouse tumors showed a significant increase in CD4+ and CD8+ T cells only in mice treated with the combination of RT+TGF xce xb2 blockade. Depletion of CD4+ or CD8+ T cells abrogated the therapeutic benefit of RT+TGF xce xb2 blockade. These data identify TGF xce xb2 as a master inhibitor of the ability of RT to generate an in situ tumor vaccine which supports testing inhibition of TGF xce xb2 during radiotherapy to promote therapeutically effective anti-tumor immunity. We used genome-wide microarray to depict main biological processes responsibles for the therapeutic benefit of the combination ofTGF-beta blockade and local radiotherapy. To gain a more comprehensice protrait of the effects of RT and TGFbeta blockade on gene expressionin tumors we collected 4T1 tumors 4 days after completion of RT. Three tumors from each group were then subjected to RNA extraction and hybridization on affymetrix array.

['"Next-generation sequencing analysis of circulating micro-RNA expression in response to parabolic flight as a spaceflight analogue"']

['"Understanding physiologic reactions to weightlessness is an indispensable requirement for safe human space missions. While adaptations of human organ systems in response to weightlessness have been described in former studies, their molecular background needs further elucidation. The study aims to analyse changes in the expression of circulating miRNAs in serum in response to gravitational changes induced by parabolic flight as a spaceflight analogue."']

Dynamic gene expression response to altered gravity in human T cells (sounding rocket flight)

We investigated differentially regulated genes in human Jurkat T lymphocytic cells in 20s and 5min microgravity and in hypergravity and compared expression profiles to identify potential gravity-regulated genes and adaptation processes. This dataset is part of a series of three and the other two datasets are deposited in GLDS-172 and GLDS-189.

Gene expression profiles of irradiated lung tissue in three mouse strains

The aim of our research is to clarify the mechanisms generating heterogeneity in response to C-ion irradiation that arise from individual genetic variations in humans. In this study we performed whole lung C-ion irradiation using three different strains of mice to examine whether strain-dependent differences in radiation effects occur in high-LET C-ion thoracic irradiation. Murine strain-variance was evaluated by histopathological examination of intra-alveolar hemorrhage that is likely to occur during the early phase after irradiation and of lung fibrosis during the late phase occurring more than three months after irradiation. We also performed microarray analysis to identify the key genes that are differentially regulated in different mouse strains after C-ion irradiation and to determine the mechanism of strain-dependent pulmonary damage after high-LET C-ion irradiation. Eight-week old female inbred C3H/He Slc C57BL/6J Jms Slc and A/J Jms Slc mice (3 kinds of mice) were used. The whole thorax of three mice was locally irradiated at 10 Gy with C-ion beams with a reference beam 137Cs gamma-rays (2 kinds of beam). Three mice of each strain (3 mice) were sacrificed and immediately dissected for lung extraction at 6 h (6h sample). Lungs for three mice of each strain without irradiation were extracted at the same time as control samples (control sample). Hybridization to microarrays (Whole Mouse Genome 4x44K OligoMicroarray Kit single color Cyamine 3-CTP) consisting of 44,000 total spots was conducted using an Agilent Gene Expression hybridization kit. The arrays from three independent replicates for each sample (3 arrays) were scanned on an Agilent dual-laser Microarray Scanner (all from Agilent Technologies) following the manufacturer s instructions. A total of 108 samples were analyzed. 3 replicate arrays x 3 mice x 3 strains x 2 timing for sampling x 2 kinds of beam.

Impact of spaceflight on gene expression in the thymus

The thymus undergoes atrophy during spaceflight. In this study we analyzed gene expression of the thymus of mice on board International space station to elucidate molecular aspects of the thymic atrophy by spaceflight.

Effect of simulated microgravity on E. coli K12 MG1655 growth and gene expression

This study demonstrates simulated microgravity effects on E. coli K 12 MG1655 when grown on LB medium supplemented with glycerol. The results imply that E. coli readily reprograms itself to combat the multiple stresses imposed due to microgravity. Under these conditions it survives by upregulating oxidative stress protecting genes and simultaneously down regulating the membrane transporters and synthases to maintain cell homeostasis. In this study a clinostat that mimics microgravity conditions was used to investigate the effects of microgravity on E. coli grown in LB medium supplemented with glycerol to monitor the effects on growth and global gene expression using Affymetrix DNA microarrays.

['Spaceflight Modulates Gene Expression in Astronauts']

['Astronauts are exposed to a unique combination of stressors during spaceflight, which leads to alterations in their physiology and potentially increases their susceptibility to infectious pathogens. Here we report the first microarray evaluation of any astronaut tissue sample, specifically whole blood, before and after spaceflight using an array comprising 234 well-characterized stress response genes. Differentially regulated genes included those important for DNA repair, oxidative stress, and protein folding/degradation. Microarrays comprising 234 well characterized stress-related genes were used to profile transcriptomic changes in six astronauts before and after short-duration spaceflight. Blood samples were collected for analysis from each eastronaut 10 days prior and 2-3 hours after return from spaceflight. Data submitted for platform GPL140 contain genes that have been pre-filtered by the analytical software to remove values of low certainty, resulting in missing values for some samples. Unfortunately, these original data are no longer available due to physical damage at Tulane University during hurricane Katrina, but the processed values were retained in redundant locations and these are submitted for upload to GEO.']

Radiation-induced bystander effects and gene expression in cells deficient for RAD9

Background: The radiation bystander response is an important component of the overall response of cells to radiation and critical to understanding health risks of radiation exposure to humans. The mechanism of radiation response includes inter-cellular signaling and intra-cellular communication by which the bystander signal is propagated. Methods: We measured the bystander response to 1Gy a-particle radiation in Mrad9-/- mouse stem cells and H1299shRAD9 cells using chromosomal aberration and micronucleus formation as DNA damage endpoints. In the H1299 model we used whole genome microarray analyses to profile the transcriptome of irradiated and bystander cells. Results: We investigated the role of RAD9 in the bystander response and showed that depletion or mutation of RAD9 had an effect of increasing chromosomal structural damage as well as micronucleus formation in bystander cells. The enhancement of the damage effect correlated strongly with a transcriptomic response in critical pathways. RAD9 depletion affected many pathways in the cell including the UV-MAPK pathway involving p38MAPK members STAT1 and PARP1 at the mRNA levels. There was an overall reduction of RNA biogenesis of gene members of this pathway suggesting that perhaps these signaling pathways do not function optimally after RAD9 depletion. Using network analysis we found there may be differential activation of transcriptional regulators between the irradiated and bystander cells involving the SP1 and NUPR1 transcription factors. Network analysis also suggested that HIF1a (Hypoxia induced factor 1a) activation could be a negative predictor of the bystander effect and perhaps that local hypoxic stress observed by cells that are directly exposed to radiation may predict whether or not they will elicit a bystander response. Gene expression in H1299 cells was measured at 4 hours after exposure to 1 Gy a-particles. There were two groups based on RAD9 status RAD9 normal and RAD9 depleted by siRNA. In each of these groups sham irradiated direct irradiated cells for positive bystanders positive bystanders direct irradiated cells for negative bystanders and negative bystanders; were identified based on micronucleus responses. Five biological replicates were analyzed for each experimental group.

Expression data from rice calli cultured with 2,4-D

Four days old rice calli were selected to grow 324h under spaceflight controls 1g-flight controls and 1g-ground controls. We used microarrays to detail the global programme of gene expression underlying cellularisation and identified distinct classes of up-regulated genes during this process. Rice calli were selected at different treatment for RNA extraction and hybridization on Affymetrix microarrays. We sought to obtain the diff-genes that caused by the microgravity.

A study of gene expression influenced by simulated microgravity in human lymphoblastoid cells

Here in this study we systematically examined the patterns of DNA methylation and hydroxy-methylation with its functional implications in gene regulation for the cultured TK6 lymphoblastoid cells upon exposure to micro-gravity conditions. The results reported here indicate that simulated microgravity alters methylation patterns in a limited way and subsequently the expression of genes involved in stress response like ATF3 FBXO17 MAP3K13 and VCL in TK6 cells. Overall design: Examination of RNA-seq with 2 replicates each for 1 cell type

Dynamic gene expression response to altered gravity in human myelomonocytic U937 cells [parabolic]

We investigated differentially regulated genes in human myelomonocytic U937 cells in 20s and 5min microgravity and in hypergravity and compared expression profiles to identify potential gravity-regulated genes and adaptation processes. Overall design: Human myelomonocytic U937 cells were exposed to altered gravity during a parabolic flight.

Dynamic gene expression response to altered gravity in human T cells (parabolic flight)

Expression Data from International C.elegans Experiment 1st

The effect of microgravity on gene expression in C.elegans was comprehensively analysed by DNA microarray. This is the first DNA microarray analysis for C.elegans grown under microgravity. Hyper gravity and clinorotation experiments were performed as reference against the flight experiment.

Comparison of gene expression profiles of normal human bronchial epithelial cells in 2D and 3D cultural conditions

The experiment is part of a project to study DNA repair process after ionizing radiation in organotypic 3-dimentional human bronchial epithlial cell culture. Human bronchial epithelial cells were grown in tissue culture flask (2D) or in matrics gel (3D). Three independent cultures were done for each condition.

['Inflammatory gene expression signatures in idiopathic intracranial hypertension: possible implications in microgravity-induced ICP elevation - Plasma']

['The visual impairment and intracranial pressure (VIIP) syndrome is a neuro-ophthalmologic condition described in astronauts returning from long duration space missions. Idiopathic intracranial hypertension (IIH), also known as pseudotumor cerebri, is characterized by a chronic elevation of intracranial pressure (ICP) in the absence of an intracranial mass lesion. Because VIIP and IIH share some neurologic and ophthalmologic manifestations, the latter might be used as a model to study some of the processes underlying VIIP. This work constitutes a preliminary investigation of the molecular pathways associated with the elevation of ICP in IIH. Gene expression signatures were obtained from exosomes collected from CSF and plasma in patients with possible signs of IIH. The gene expression targets focused on inflammatory genes and miRNAs. The results suggest that inflammatory cytokine-driven processes and immune cell migration are activated when ICP is elevated in IIH patients, either as a cause or effect of the ICP increase. Several miRNAs appear to be involved in this response, among which miR-9 and miR-16 are upregulated in CSF and plasma of higher ICP subjects. This study provides evidence in support of neurophysiological alterations and neuro-immunomodulation in this condition. If similar changes are seen in astronauts manifesting with the VIIP syndrome, an underlying pathophysiological basis may be discovered.']

Global gene expression analysis highlights microgravity sensitive key genes in longissimus dorsi and tongue of 30 days space-flown mice

Microgravity as well as chronic muscle disuse are two causes of low back pain originated at least in part from paraspinal muscle deconditioning. At present no study investigated the complexity of the molecular changes in human or mouse paraspinal muscles exposed to microgravity. The aim of this study was to evaluate longissimus dorsi and tongue (as a new potential in-flight negative control) adaptation to microgravity at global gene expression level. C57BL/N6 male mice were flown aboard the BION-M1 biosatellite for 30 days (BF) or housed in a replicate flight habitat on ground (BG). Global gene expression analysis identified 89 transcripts differentially regulated in longissimus dorsi of BF vs. BG mice (False Discovery Rrate < 0,05 and fold change < -2 and > +2) while only a small number of genes were found differentially regulated in tongue muscle ( BF vs. BG = 27 genes). Overall Design: C57BL/N6 mice were randomly divided in 3 groups: Bion Flown (BF) mice flown aboard the Bion M1 biosatellite in microgravity environment for 30 days; Bion Ground (BG) mice housed in the same habitat of flown animals but exposed to earth gravity; and Flight Control (FC) mice housed in a standard animal facility.

Chronic low dose-rate versus acute ionizing radiation induced gene expression in primary derived prostate fibroblasts

Using a novel flood source construct we performed a direct comparison of genome-wide gene expression regulations resulting from exposure of primary human prostate fibroblast cultures to acute (10cGy and 200cGy) and longer-term chronic (1.0-2.45cGy cumulative over 24hr) exposures.