Gene expression changes induced by ILB in Schwann like cells
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ABSTRACT:
PROVIDER: PRJNA641711 | ENA |
REPOSITORIES: ENA
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Project description:To understand the mechanism of action of ILB we have used gene expression analysis to document the changes induced by the drug in Schwann like cells. Human Schwann-like cells (ATCC-CRL-2884) were treated with ILB (0.01 mg/ml) for 48 hours. Control samples were cultured parallel with no drug added. The expression data was used to identify the ILB-regulated genes.
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Project description:We used microarrays to detail the transcriptome-wide gene expression changes underlying chemical conversion of human fibroblasts into induced Schwann Cells over a time period of 39 days. We compared then the expression profiles of these induced Schwann Cells to primary Schwann cells. The gene expression results analyzed in this study are further described in Thoma et al. (2014) Chemical conversion of human fibroblasts into functional Schwann cells. Under submission
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Project description:Schwann cells are critical for the proper development and function of the peripheral nervous system, where they form a collaborative relationship with axons. Past studies highlighted that a pair of proteins called the prohibitins play major roles in Schwann cell biology. Prohibitins are ubiquitously expressed and versatile proteins. We have previously shown that while prohibitins play a crucial role in Schwann cell mitochondria for long-term myelin maintenance and axon health, they may also be present at the Schwann cell-axon interface during development. The current data set uses BioID to identify a pool of candidate PHB2 interactors in Schwann cells and explores how the PHB2 interactome changes depending on neuronal signals. We expressed a PHB2-turboID fusion construct in primary rat Schwann cells; turboID is a 35 kDa engineered biotin ligase that rapidly biotinylates proximal proteins. Thus, proteins interacting with PHB2-turboID within an approximately 10 nm radius are tagged with biotin. As a control, primary rat Schwann cells were transfected with an unfused turboID construct (Con-turboID). Schwann cells expressing Con-turboID or PHB2-turboID were plated alone or onto primary rat DRG neurons, in the presence or absence of biotin. After 2 hours, proteins were harvested from the cultures and biotinylated proteins were purified using streptavidin-affinity purification (AP). We then used liquid chromatography-mass spectrometry (LC-MS) to identify the PHB2-turboID interactors (the biotinylated proteins) in our purified pool. The PHB2 interactors identified here, especially those which increase or decrease interaction with PHB2-turboID in the presence of neurons, may play a role in prohibitin-associated Schwann cell-axon communication.
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Project description:We report the transcriptome analysis of HEI-286 human Schwann cells and MiaPaCa-2 pancreatic cancer cells Nerves stimulate cancer progression for different cancer types, promoting both tumor growth and perineural invasion, a prominent process in pancreatic ductal adenocarcinoma. Yet little is known about how cells in the nerves contribute to cancer progression. Here we demonstrate that Schwann cells, the most abundant cell type in nerves, collectively function as tumor activated Schwann cell tracks (TASTs). Schwann cells in a 3D matrix organize into stellate tracks which serve as a conduit for cancer cell migration. TASTs actively promote pancreatic cancer dissemination during perineural invasion and neoneurogenesis. This key function of TASTs is induced by cancer cells triggering c-Jun activation of Schwann cells, analogous to the reprogramming that promotes nerve repair. In TASTs, dynamic Schwann cell processes wrap around and apply forces directly on cancer cells to enhance cancer motility. We define a model of cancer progression that relies upon collective Schwann cell behavior driven by c-Jun transcriptional reprogramming.
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Project description:To obtain the dynamic gene expression of myelinating Schwann cells, we have employed gene expression profiling microarray as a discovery platform to analyze the gene expression of Schwann cells in different stages of myelination in an DRG neuron and SC co-culture myelinating model. Rat Schwann cells and dorsal root ganglion (DRG) neurons were cocultured and induced myelination in DMEM medium containing 15% FBS, 50 ng/ml NGF and 50 μg/ml L-ascorbic acid for 21d. During the co-cultivation, myelinating SCs at different stages dissected by Laser microdissection (LMD) in myelination model (i.e. co-culture 1d, 3d, 7d, 14d, 21d), the Schwann cells without co-culture as control samples (i.e. co-culture 0d). The results from Euclidean distance matrix, principal component analysis, and hierarchical clustering indicated that 2 nodal transitions in temporal gene expressions could segregate 3 distinct transcriptional phases within the period of DRG/SC co-culture 21 days. The 3 phases were designated as “premyelination”, “myelination”, and “mature phase”, respectively, by referring to morphological observation of post co-culture changes and gene ontology (GO) analysis.
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