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Genomic and phenotypic alterations of the neuronal-like cells derived from the human embryonal carcinoma stem cells (NT2) caused by exposure to the organophosphorus compounds paraoxon and mipafox

ABSTRACT: hNT2 under neurodifferentiation were exposed during 4 days to 1 µM of the non-neuropathic organophosphorus compound paraoxon and 5 µM of the neuropathic organophosphorus compound and afterwards transcriptomics were analysed and compared regarding control non-exposed cells Exposure to paraoxon caused alteration of 137 genes while only a single gene was altered after exposure to mipafox

ORGANISM(S): Homo sapiens

PROVIDER: GSE38050 | GEO | 2012/10/18

SECONDARY ACCESSION(S): PRJNA167130

REPOSITORIES: GEO

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Genomic and phenotypic alterations of the neuronal-like cells derived from the human embryonal carcinoma stem cells (NT2) caused by exposure to the organophosphorus compounds paraoxon and mipafox

Project description:hNT2 under neurodifferentiation were exposed during 4 days to 1 ÂµM of the non-neuropathic organophosphorus compound paraoxon and 5 ÂµM of the neuropathic organophosphorus compound and afterwards transcriptomics were analysed and compared regarding control non-exposed cells Exposure to paraoxon caused alteration of 137 genes while only a single gene was altered after exposure to mipafox Comparing two experimental conditions: control versus paraoxon or mipafox exposed cells. Three biological replicates.

2012-10-18 | E-GEOD-38050 | biostudies-arrayexpress

Dichlorvos-induced formation of isopeptide crosslinks between proteins in SH-SY5Y cells

Project description:Chlorpyrifos oxon catalyzes the crosslinking of proteins via an isopeptide bond between lysine and glutamic acid or aspartic acid in studies with purified proteins. Our goal was to determine the crosslinking activity of the organophosphorus pesticide, dichlorvos. We developed a protocol for examining crosslinks in a complex protein mixture consisting of human SH-SY5Y cells exposed to 10 µM dichlorvos. The steps in our protocol included immunopurification of crosslinked peptides by binding to anti-isopeptide antibody 81D1C2, stringent washing of the immobilized complex, release of bound peptides from Protein G agarose with 50% acetonitrile 1% formic acid, liquid chromatography tandem mass spectrometry on an Orbitrap Fusion Lumos mass spectrometer, Protein Prospector searches of mass spectrometry data, and manual evaluation of candidate crosslinked dipeptides. We report a low quantity of dichlorvos-induced KD and KE crosslinked proteins in human SH-SY5Y cells exposed to dichlorvos. Cells not treated with dichlorvos had no detectable KD and KE crosslinked proteins. Proteins in the crosslink were low abundance proteins. In conclusion, we provide a protocol for testing complex protein mixtures for the presence of crosslinked proteins. Our protocol could be useful for testing the association between neurodegenerative disease and exposure to organophosphorus pesticides.

2023-10-24 | PXD033593 | Pride

SILENCING OF PNPLA6, THE NEUROPATHY TARGET ESTERASE (NTE) CODIFYING GENE, ALTERS HUMAN NEURODIFFERENTIATION IN VITRO

Project description:Neuropathy Target Esterase (NTE) is a protein involved in the development of a polyneuropathy caused by exposure to certain organophosphorus compounds. In vivo and in vitro studies have also associated NTE with embryonic development since NTE null mice embryos are non-viable, and silencing the NTE-codifying gene (Pnpla6) in mouse embryonic stem cells strongly alters the differentiation of vascular and nervous systems. In this paper, human embryonal carcinoma stem cells (hNT2) are used as in vitro neurodifferentiation model to determine whether PNPLA6 gene silencing is able to alter the differentiation into a neuronal phenotype. In controls, the PNPLA6 mRNA levels increased in parallel with other neuroectodermal markers during the neurodifferentiation. PNPLA6 gene silencing with specific interference RNA led to a 97% maximum decrease in gene expression by day 3 after transfection, with a maximum of 50% reduction in NTE enzymatic activity by day 4. Microarray analyses of the PNPLA6-silenced cells showed alterations in several developmental processes, mainly neurogenesis and epithelium tube morphogenesis. PNPLA6 silencing also led to a reduction in electrical activity and an altered neuronal phenotype. This work is the first proof supporting the hypothesis that NTE protein plays a critical role in human early neurodevelopment using a human cell differentiation model. PNPA6 gene silencing in undifferentiatied hNT2 cells was performed, together with a Control (non-gene silencing cells) and a Negative Control (non-specific RNA silencer). Three biological replicates.

2014-10-06 | E-GEOD-55761 | biostudies-arrayexpress

Evaluation of mass spectrometry MS/MS spectra for the presence of isopeptide crosslinked peptides

Project description:Chronic low dose exposure to organophosphorus pesticides is associated with risk of neurodegenerative disease. The mechanism of neurotoxicity is unknown, though it is independent of acetylcholinesterase inhibition. Adducts on tyrosine, lysine, threonine, and serine can occur after exposure to organophosphorus pesticides, the most stable being adducts on tyrosine. Rabbit monoclonal 1C6 to diethoxyphosphate modified tyrosine (depY) was created by single B cell cloning. The amino acid sequence and binding constant (Kd 3.2 x 10-‐8 M) were determined. Cultured human neuroblastoma SH-‐SY5Y and mouse neuroblastoma N2a cells incubated with a sub-‐cytotoxic dose of 10 µM chlorpyrifos oxon contained depY-‐modified proteins detected by monoclonal 1C6 on Western blots. DepY-labeled peptides from tryptic digests of cell lysates were imunopurified by binding to immobilized 1C6. Peptides released with 50% acetonitrile, 1% formic acid were analyzed by LC-‐MS/MS on an Orbitrap Fusion Lumos mass spectrometer. Protein Prospector database searches identified 51 peptides modified on tyrosine by diethoxyphosphate in human SH-‐SY5Y cell lysate and 73 diethoxyphosphate-‐modified peptides in mouse N2a cell lysate. Adducts appeared most frequently on the cytoskeleton proteins tubulin, actin, and vimentin. It was concluded that rabbit monoclonal 1C6 can be useful for studies that aim to understand the mechanism of neurotoxicity resulting from low dose exposure t organophosphorus pesticides.

2022-02-17 | PXD027203 | Pride

SILENCING OF PNPLA6, THE NEUROPATHY TARGET ESTERASE (NTE) CODIFYING GENE, ALTERS HUMAN NEURODIFFERENTIATION IN VITRO

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Transcriptional effects of bipolar disorder drugs on NT2-N cells

Project description:Human neuronal-like cells (NT2-N) were treated with either lamotrigine (50 µM), lithium (2.5 mM), quetiapine (50 µM), valproate (0.5 mM) or vehicle control for 24 hours. Genome wide mRNA expression was quantified by RNA-sequencing. Results offer insights on the mechanism(s) of action of bipolar disorder drugs at the transcriptional level.

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Transcriptomic analysis of Mtb H37Rv on treatment with ATD-3169 (Redox generating compound)

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2014-11-30 | GSE61618 | GEO

Evaluation of Some Risk Factors Associated With Colorectal Cancer

Project description:1. Evaluation of some risk factors (e.g. smoking, heavy metal exposure, and organophosphorus exposure) on CRC. 2. Estimation of KRAS and BRAF genes mutations associated with CRC. 3. Association between different risk factors and gene mutations on different types of CRC.

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Role of SOX2 in the etiology of embryonal carcinoma, based on analysis of the NCCIT and NT2 cell lines.

Project description:The transcription factor SOX2, associated with amongst others OCT3/4, is essential for maintenance of pluripotency and self-renewal of embryonic stem cells. SOX2 is highly expressed in embryonal carcinoma (EC), the stem cell component of malignant nonseminomatous germ cell tumors, referred to as germ cell cancer (GCC). In fact, OCT3/4 together with SOX2 is an informative diagnostic tool for EC in a clinical setting. Several studies support the hypothesis that SOX2 is a relevant oncogenic factor in various cancers and recently, SOX2 has been suggested as a putative therapeutic target for early stage EC. We demonstrate the presence of genomic amplification of SOX2 in an EC cell line, NCCIT, using array comparative genome hybridization and fluorescence in situ hybridization. Down-regulation of SOX2 by targeted siRNA provokes NCCIT cells towards apoptosis, while inhibition of OCT3/4 expression induced differentiation, with retained SOX2 levels. Mice pluripotent xenografts from NCCIT (N-NCCIT and N2-NCCIT) show a consistent SOX2 expression, in spite of loss of the expression of OCT3/4, and differentiation, with retained presence of genomic amplification. No SOX2 amplification has been identified in primary pure and mixed EC in vivo patient samples so far. The data presented in this study are based on a single EC cell line with a SOX2 amplification, with NT2 as control EC cell line, showing no profound induction of apoptosis upon SOX2 downregulation. The findings are of relevance to identify mechanisms involved in the pathogenesis of EC tumors, and support the model of SOX2-oncogene dependency of EC, which however, does not exclude induction of differentiation. This finding is likely related to the presence of wild type p53 in GCC, resulting in expression of downstream target genes, amongst others miR-34a, miR-145 and SOX2, associated to the unique sensitivity of GCC to DNA damaging agents.

| S-EPMC3880257 | biostudies-literature

Gene expression changes in human cardiomyocytes exposed to VX (O-ethyl S-[2-(diisopropylamino)ethyl] methylphosphonothiolate)

Project description:Organophosphorus compounds induce cardiotoxicity through currently unknown mechanisms, which need to be unraveled by a comprehensive and systematic approach such as genome-wide gene expression analysis. We used microarrays to study gene expression changes in human cardiomyocytes after exposure to VX, and identified pathways underlying these changes. Human cardiomyocytes were exposed to sublethal concentrations (0, 0.1, 10 μM) of VX. RNA were extracted at different timepoints (0, 6, 24, 72 h) after VX exposure and hybridized to Affymetrix microarrays. Four biological repeats were used for each condition.

2011-10-28 | E-GEOD-33325 | biostudies-arrayexpress

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