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A 5’ Sequence of Human HMGB2 Gene for Transcriptional Targeting of Glioblastoma

ABSTRACT: Achievement of specific tumor cell targeting remains a challenge for glioma gene therapy. We report here the identification and characterization of a 5’ sequence of human HMGB2 gene for transcriptional targeting to glioblastoma. We performed microarray analysis and found HMGB2 as one of the genes that had a low level of expression in normal human astrocytes, but was significantly up-regulated in glioblastoma cells. Real-time PCR quantification revealed increase in HMBG2 expression level in glioblastoma tissues and cells between 11 to 79 fold over that in normal human brain tissue. With progressive truncation of a 5’-upstream sequence of the HMGB2 gene, we identified a 500-bp fragment that displayed a high transcriptional activity in glioblastoma cells, but a low activity in normal brain cells. Using the sequence to drive the expression of the herpes simplex virus thymidine kinase gene in the context of a baculoviral vector, glioblastoma cells died in the presence of ganciclovir, whereas normal human astrocytes and neurons were not affected. We further confirmed that after intra-tumor injection, the baculoviral vector effectively suppressed the growth of human glioblastoma cells in a mouse xenograft model. Our results suggest that the 5’-upstream sequence of the HMGB2 gene can be used as an efficient, tumor-selective promoter in targeted vectors for glioblastoma gene therapy. U251 cells (n=3) genes level expression were compared to that of normal astrocytes (n=3) to find overexpressed genes in glioblastoma. Highly expressed genes were compared to those found in the litterature. This was selected to clone promoters of highly expressed genes in glioblastomas

ORGANISM(S): Homo sapiens

SUBMITTER: Jerome Boulaire

PROVIDER: E-GEOD-12305 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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A 5’ Sequence of Human HMGB2 Gene for Transcriptional Targeting of Glioblastoma

Project description:Achievement of specific tumor cell targeting remains a challenge for glioma gene therapy. We report here the identification and characterization of a 5’ sequence of human HMGB2 gene for transcriptional targeting to glioblastoma. We performed microarray analysis and found HMGB2 as one of the genes that had a low level of expression in normal human astrocytes, but was significantly up-regulated in glioblastoma cells. Real-time PCR quantification revealed increase in HMBG2 expression level in glioblastoma tissues and cells between 11 to 79 fold over that in normal human brain tissue. With progressive truncation of a 5’-upstream sequence of the HMGB2 gene, we identified a 500-bp fragment that displayed a high transcriptional activity in glioblastoma cells, but a low activity in normal brain cells. Using the sequence to drive the expression of the herpes simplex virus thymidine kinase gene in the context of a baculoviral vector, glioblastoma cells died in the presence of ganciclovir, whereas normal human astrocytes and neurons were not affected. We further confirmed that after intra-tumor injection, the baculoviral vector effectively suppressed the growth of human glioblastoma cells in a mouse xenograft model. Our results suggest that the 5’-upstream sequence of the HMGB2 gene can be used as an efficient, tumor-selective promoter in targeted vectors for glioblastoma gene therapy.

2008-12-31 | GSE12305 | GEO

Expression data of the splenocytes from mice at 6th of Plasmodium chabaudi infection.

Project description:During malaria infection is observed a robust immune response culminating on release of inflammatory mediators. This exacerbated immune response is involved in malaria symptoms and mortality. There are evidences that this response is mediated by innate immunity where pattern recognition receptors have a key role. We used microarrays to elucidate some pro-inflammatory genes that are differential expressed during P. chabaudi infection, a malarial murine model Spleen from C57BL/6 or MyD88 knockout mice non-infected or after 6 days post infection with 105 P. chabaudi infected red blood cells were harvested for RNA extraction and hybridization on Affymetrix microarrays. This time point was chose to coincides with rupture of red blood cells \since this event of the parasite life cycle is related with malaria outcomes.

2012-01-13 | E-GEOD-35083 | biostudies-arrayexpress

Gene Expression Changes in Response to UV Inactivated Baculoviral Vector Transduction of the Rat Brain In Vivo

Project description:Recombinant baculoviral vectors efficiently transduce several types of cells in the brain. To characterize host responses to viral challenge, thus verifying the suitability of using the virus for the development of gene therapy strategies in the central nervous system, we used cDNA microarray technology to examine in vitro and in vivo global cellular gene expression profiles after viral transduction. We demonstrated that the transduction induced host antiviral responses as a major reaction in all three types of samples profiled, including the rat brain, cultured human astrocytes and human neuronal cells. The related genes were mainly those associated with innate immunity. Several genes of the major histocompatibility complex molecules, an important component of the host adaptive immunity to exogenous pathogens, were up-regulated in the rat brain and human astrocytes, but not in neuronal cells. We also observed that genes related to cell death and apoptosis were up-regulated and genes related cell cycle regulation were down-regulated in neuronal cells, but not obviously affected in astrocytes. These findings should be useful in understating the molecular basis for neural cell response to baculoviral transduction and guiding rational applications of baculoviral vectors in the central nervous systems Keywords: Baculovirus, brain, astrocytes, neurons, transduction, Infection, gene expression analysis, DNA microarray

2009-04-17 | GSE14484 | GEO

Gene Expression Changes in Response to Baculoviral Vector Transduction of Normal Human Astrocytes In Vitro

2007-12-13 | GSE9834 | GEO

Gene Expression Changes in Response to Baculoviral Vector Transduction of the Rat Brain In Vivo

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Gene Expression Changes in Response to Baculoviral Vector Transduction of Neuronal Cells In Vitro

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RNA-seq of glioma and normal cell lines after HDAC1 knock-down

Project description:Using RNA-seq we characterized the effect of shRNA mediated knockdown of HDAC1 in glioblastoma stem cells (GSCs), normal ihNPCs, and normal human astrocytes.

2021-07-14 | GSE179882 | GEO

Transcriptional changes in glioma cells following treatment with 1-Benzyl-3-cetyl-2-methylimidazolium iodide (NH125), Tunicamycin, and 0.1% DMSO (in media)

Project description:Purpose: Evaluate transcriptional changes in glioma derived stem cells, U251, and normal human astrocytes following treatment with NH125, Tunicamycin, or 0.1% DMSO Methods: Glioma derived stem cells, U251 and Normal Human Astrocytes were plated in individual dishes and treated with either 2.5 micromolar NH125, 1.0 mcg/mL Tunicamycin, or 0.1% DMSO (vehicle) for twenty-four hours followed by total RNA extraction, library preparation and next generation sequencing Results: Using our analysis pipeline we mapped our sequence reads to the reference genome GRCh38. Following read count normalization and differential expression of NH125 and tunicamycin treated samples to vehicle treated controls we found that NH125 and tunicamycin treatment lead to activation of similar signalling pathways Conclusions: We present an optimized workflow for analysis of transcriptional changes in drug treated glioma derived stem cells, glioblastoma cells and normal human astrocytes

2018-03-02 | GSE102505 | GEO

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Project description:Gene Expression Changes in Response to Baculoviral Vector Transduction of Normal Human Astrocytes In Vitro

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Expression data from glioblastoma stem-like cells and their differentiated progeny co-cultured with astrocytes

Project description:Glioblastoma stem-like cells or their differentiated progeny were co-cultured for 48h with normal human astrocytes to detect if invasion associated genes were influenced

2012-04-09 | GSE37120 | GEO

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