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Differential gene expression in Lithium chloride (LiCl) treated Scleroderma (SSc) skin fibroblasts

ABSTRACT: Differential gene expression in Lithium chloride (LiCl) treated Scleroderma (SSc) skin fibroblasts

PROVIDER: PRJNA1071564 | ENA |

REPOSITORIES: ENA

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Differential gene expression in Lithium chloride (LiCl) treated Scleroderma (SSc) skin fibroblasts

Project description:Human scleroderma skin derived fibroblast were treated with LiCL. This is a study of differential gene expression between LiCl treated SSc group and untreated SSc group.

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The effects of antimigratory drugs lithium chloride and 6-bromoindirubin-3'-oxime (BIO) in glioblastoma cells

Project description:The purpose of the experiment was to better understand the effects of antimigratory drugs lithium chloride and 6-bromoindirubin-3'-oxime (BIO) in glioblastoma cells. U251 cells were grown to 60-70% confluency in DMEM containing 10% FBS in 6 well plates. Drugs were added (LiCl 20 mM, BIO 5 micromolar, with carrier controls; either NaCl or DMSO) and incubated for 24 hours. Cells were then harvested and RNA purified using TRIZOL.

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Transcription profiling of galactose-grown S. cerevisiae cells exposed to lithium chloride

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Real-time quantitative PCR analysis of 88 microRNAs involved in human cell differentiation and development in involved skin of systemic scleroderma (SSc), localized scleroderma (LSc) and Keloid.

Project description:Skin specimens were derived from involved skin of 3 systemic scleroderma (SSc), 3 localized scleroderma (LSc) and 3 keloid patients. These skin samples and 3 control skins were collected and fixed in formaldehyde immediately after resections. The microRNA (miRNA) isolation from human skin tissue was performed using miRNeasy FFPE kit (Qiagen). For PCR array, miRNAs were reverse-transcribed into first strand cDNA using RT2 miRNA First Strand Kit (SABiosciences). A mixture of equal amounts of miRNAs from 3 normal skins, 3 SSc, 3 LSc or 3 keloid were prepared, and miRNA expression profile in each disease in vivo was evaluated using RT2 Profiler PCR Array. The cDNA was mixed with RT2 SYBR Green/ROX qPCR Master Mix and the mixture was added into 96-well RT2 miRNA PCR Array that includes primer pairs for 88 human miRNAs (SABiosciences). Skin specimens were derived from involved skin of 3 systemic scleroderma (SSc), 3 localized scleroderma (LSc) and 3 keloid patients. These skin samples and 3 control skins were collected and fixed in formaldehyde immediately after resections. Control donors were each matched with diseases for age, sex, and biopsy site.

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DNA methylation array of irradiated and/or lithium exposed iPSC-derived human neural progenitor cells

Project description:DNA isolation was performed following the DNeasy Blood and Tissue Kit (Qiagen). Cells were irradiated in an CIX2 Xstrahl X-Ray Irradiator with Aluminum filter at a focus-to-skin distance (FSD) of 40 cm and with an energy of 195kV, 10 mA, for and absorbed dose of 4 GY. To allow for assessment of irradiation damage, cells remained in the incubator for 1 hour. Pre-warmed DMEM/F-12 was mixed with the appropriate amount of LiCl powder (Sigma-Aldrich, Cat# L9650-100G), passed through a 0.22 μm filter, and supplemented with 1% Penicillin-Streptomycin, 1% N2, 0.1% B27, 0.1% EGF, and 0.1% FGF2 to produce 3mM Lithium Chloride (LiCl) hNSPC medium.

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Real-time quantitative PCR analysis of 88 microRNAs involved in human cell differentiation and development in involved skin of systemic scleroderma (SSc), localized scleroderma (LSc) and Keloid.

2013-01-15 | GSE43469 | GEO

Gene expression array of irradiated and/or lithium exposed iPSC-derived human neural progenitor cells

Project description:RNA isolation was performed using the RNeasy Mini Kit (Qiagen), per manufacturer instructions. For differential gene expression analysis, the Affymetrix Whole Transcript Assay with the Clariom S cartridge for human samples (n=2, BEA core facility, Sweden) was used. Cells were irradiated in an CIX2 Xstrahl X-Ray Irradiator with Aluminum filter at a focus-to-skin distance (FSD) of 40 cm and with an energy of 195kV, 10 mA, for and absorbed dose of 4 GY. To allow for assessment of irradiation damage, cells remained in the incubator for 1 hour. Pre-warmed DMEM/F-12 was mixed with the appropriate amount of LiCl powder (Sigma-Aldrich, Cat# L9650-100G), passed through a 0.22 μm filter, and supplemented with 1% Penicillin-Streptomycin, 1% N2, 0.1% B27, 0.1% EGF, and 0.1% FGF2 to produce 3mM Lithium Chloride (LiCl) hNSPC medium.

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Fli1 downregulation in scleroderma myeloid cells has profibrotic and proinflammatory effects

Project description:Scleroderma is an autoimmune connective tissue disease characterized by immune dysregulation, vasculopathy and fibrosis. We have previously demonstrated that low Fli1 expression in scleroderma fibroblasts and endothelial cells plays an important role in SSc pathogenesis. Cells of myeloid and lymphoid origin also express Fli1 and are dysregulated in patients with SSc, playing key roles in disease pathogenesis However, a role for immune Fli1 in SSc has not yet been described. Our aim was to elucidate whether Fli1 contributes to the immune dysregulation seen in scleroderma. Comparison of the expression of Fli1 in monocytes, B- and T-cell fractions of PBMCs isolated from scleroderma patients and healthy controls, showed an increase in Fli1 levels in monocytes. We used siRNA transfected human myeloid cells and mouse peritoneal macrophages obtained from Fli1flox/floxLysMCre+/+ mice, and found that markers of alternative macrophage activation were increased with Fli1 deletion. Coculture of Fli1-deficient myeloid cells and primary human or mouse fibroblasts resulted in a potent induction of collagen type I, independent of TGFβ upregulation. We next analyzed global gene expression profile in response to Fli1 downregulation, to gain further insight into the molecular mechanisms of this process and to identify differentially expressed genes in myeloid cells. Of relevance to SSc, the top most upregulated pathways were hallmark IFN-ɣ and IFN-α response. Additionally, several genes previously linked to SSc pathogenesis and fibrosis in general were also induced, including MCP-1, MCP-3, MMP12, and CXCL10. ANKRD1, a profibrotic transcription co-regulator was the top upregulated gene in our array. Our results show that Fli1-deficient myeloid cells share key features with cells from SSc patients, with higher expression of profibrotic markers and activation of interferon responsive genes, thus suggesting that low Fli1 may contribute to SSc pathogenesis.

2020-06-09 | GSE144625 | GEO

Scleroderma Morphea Normal Fibroblasts

Project description:Fibroblast were isolated from either back or forearm punch biopsies taken by Dr. Kari Connolly(UCSF) from either normal control patients, patients with a diagnosis of Systemic Sclerosis with Diffuse Scleroderma and patients with a diagnosis of morphea. Scleroderma fibroblast were grown to confluence, media was changed and the RNA prepared from the cells 48 hrs later (to avoid serum responsive genes). A disease state experiment design type is where the state of some disease such as infection, pathology, syndrome, etc is studied. Keywords: disease_state_design

2005-12-21 | GSE3886 | GEO

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