ABSTRACT: Expression data from normal human fibroblasts expressing prelamin A or progerin, untreated or treated with farnesyltransferase inhibitor (FTI)
Project description:We compared the transcriptomes of isogenic diploid fibroblasts expressing progerin or elevated levels of wild-type prelamin A with that of wild-type fibroblasts. We subsequently used the reversion towards normal of two phenotypes, reduced cell growth and dismorphic nuclei, by treatment with farnesyltransferase inhibitor (FTI) or overexpression of ZMPSTE24, as a filtering strategy to identify genes linked to the onset of these two phenotypes.
Project description:We compared the transcriptomes of isogenic diploid fibroblasts expressing progerin or elevated levels of wild-type prelamin A with that of wild-type fibroblasts. We subsequently used the reversion towards normal of two phenotypes, reduced cell growth and dismorphic nuclei, by treatment with farnesyltransferase inhibitor (FTI) or overexpression of ZMPSTE24, as a filtering strategy to identify genes linked to the onset of these two phenotypes. We carried out microarray analyses of gene expression profiling in isogenic fibroblasts lines to identify the subset of genes whose expression patterns are strongly altered upon expression of progerin or elevated levels of wild-type lamin A. A filtering strategy was then used to identify potential key effectors. We searched for genes whose expression was reverted towards normal by treatment with farnesyl transferase inibitors (FTI) in both cell lines for 48 hours, or by overexpression of ZMPSTE24 in cells with elevated levels of prelamin A, conditions that improve cell proliferation and leads to a significant decrease in nuclear membrane abnormalities.
Project description:We analyzed and compared global gene expression changes in fibroblasts from human subjects with HGPS compared to age matched controls. We then treated both control and HGPS fibroblasts with a protein farnesyltransferase inhibitor (FTI), a medication currently used in clinical trials to treat HGPS, to look for a reversal of the gene defects present in HGPS fibroblasts.
Project description:Human BJ fibroblasts were treated with FLI-06 and gene expression was compared to untreated fibroblasts. RNA-seq data comprises 2 groups: treated and untreated BJ fibroblasts. Jena Centre for Systems Biology of Ageing - JenAge (www.jenage.de)
Project description:Transcriptional profiling of human BJ fibroblasts comparing control FF shRNA expressing cells vs. BRD7 shRNA expressing cells under two conditions, either untreated or treated with 8uM nutln-3a for 8 hours. This experiment was done using two independent shRNAs targeting BRD7. Nutlin-3a was used to stabilize p53 and induce its transcriptional activity.
Project description:Hutchinson-Gilford progeria syndrome (HGPS) is a rare and fatal human premature aging disease1-5, characterized by premature atherosclerosis and degeneration of vascular smooth muscle cells (SMCs)6-8. HGPS is caused by a single-point mutation in the LMNA gene, resulting in the generation of progerin, a truncated mutant of lamin A. Accumulation of progerin leads to various aging-associated nuclear defects including disorganization of nuclear lamina and loss of heterochromatin9-12. Here, we report the generation of induced pluripotent stem cells (iPSCs) from fibroblasts obtained from patients with HGPS. HGPS-iPSCs show absence of progerin, and more importantly, lack the nuclear envelope and epigenetic alterations normally associated with premature aging. Upon differentiation of HGPS-iPSCs, progerin and its associated aging consequences are restored. In particular, directed differentiation of HGPS-iPSCs to SMCs leads to the appearance of premature senescent SMC phenotypes associated with vascular aging. Additionally, our studies identify DNA-dependent protein kinase catalytic subunit (DNAPKcs) as a component of the progerin-containing protein complex. The absence of nuclear DNAPKcs correlates with premature as well as physiological aging. Since progerin also accumulates during physiological aging6,12,13, our results provide an in vitro iPSC-based model with an acceleration progerin accumulation to study the pathogenesis of human premature and physiological vascular aging. Microarray gene expression profiling was done to: (1) Compare differences between WT fibroblasts and fibroblasts from patients suffering of the Hutchinson-Gilford progeria syndrome (2) Check that iPSC originating from WT and patients are in fact similar to ESC
Project description:LNCaP cells are an established androgen receptor expressing prostate carcinoma cell line. Human foreskin fibroblasts also expressing the androgen receptor were obtained from phenotypic normal male individuals. Cells were cultured either at confluency leading to G0 cell cycle state or while they were proliferating. Cells were either untreated, or treated with dihydrotestosterone (DHT) or ethanol (ETOH) which also served as the solvent for the DHT. All experimental RNA samples derived from the untreated or treated cell lines were hybridized on cDNA arrays against a common reference. This reference was composed out of common reference CRG (50%) and out of fibroblast RNA (50%). This reference is also called "mixed reference" in the description of the 26 individual experiments.
Project description:To understand the molecular mechanisms that underpin pro-tumourigenic functions of cancer-associated fibroblasts (CAFs) we compared the proteome, acetylome, phosphoproteome of human immortalised breast cancer CAFs with those of the normal mammary fibroblasts that they were generated from. Based on the results obtained, we have also analysed proteome changes when CAFs were treated with the P300/CBP inhibitor c646.
Project description:Disassembly of the nuclear envelope is an essential feature of mammalian cell division controlled by the phosphoryaltion of lamins via cyclin dependent kinases. This process is affected in cells expressing progerin, a lamin A allele found in patients with Hutchinson-Gilford Progeria syndrome. Progerin can inhibit cell proliferation of both normal and tumor cells and this property is largely magnified if its phosphorylation at serine 22 is inhibited by a genetic mutation to generate S22A-progerin. Surprisingly, S22A-progerin acquires the ability to trigger cellular senescence in tumor cells with mutations in the p53 and RB tumor suppression pathways suggesting a novel pathway to control the growth of malignant tumors. We used microarrays to characterize the gene expression changes induced by the S22A-progerin in comparison with the wt-progerin in U-2 OS cell line. U-2 OS cells were infected with a retroviral vector that express S22A-progerin or wt-progerin. After 4 days of infection, RNA was extracted from three independents replicas of each condition. Total RNA was send to Genome Quebec service for hybridization with Affymetrix microarrays.
