Project description:Immune checkpoints blockade therapy may have clinical symptoms of hyper-progressiveness (HP). The molecular mechanism of HP is still to be revealed. This study inspected transcriptome difference between clinical hyper-progressive samples to immunotherapy-response samples based public datasets. Spliceosome factors SRSF2 were found most enriched in neoantigen-related splicing events. Similar splicing pattern and increased cell invasion were found in two lung cancer cell lines after SRSF2 over-expressed.This study shows that dysregulation of SRSF2 -related splicing might be an important feature of immune hyper-progress occurrence.
Project description:We showed that the over-expression of the SR protein SRSF2 led to the regulation of transcript abundance of many genes. H358 cell line with 6 samples with SRSF2 over-expression and 6 samples as control.
Project description:To investigate the molecular mechanisms through which SRSF2 acts it was depleted in 3 epithelial cell lines; primary human keratinocytes (HK) and two squamous cell carcinoma cell lines SCC25 and FaDu cells. Additionally FaDu cells were used to make clones with stably reduced SRSF2 levels in order to explore how cells can compensate reduced SRSF2 levels. Due to SRSF2's role in transcription Cut&Run using an antibody specific for total Pol II was performed in ctr and SRSF2 depleted FaDu cells.
Project description:Mutations within genes encoding spliceosomal proteins are the most common class of mutations in patients with myelodysplastic syndromes, yet it is currently not well understood how these mutations impact hematopoiesis or RNA splicing. Here we report that mutations affecting the splicing factor SRSF2 alter its normal RNA recognition activity, resulting in impaired hematopoietic differentiation and myelodysplasia. Commonly occurring SRSF2 mutations impaired wildtype SRSF2’s normal RNA-binding avidity and preference for specific exonic splicing enhancer RNA motifs. Integration of murine and human transcriptome data identified recurrent mis-splicing of key transcriptional regulators in the presence of mutant SRSF2, including promotion of a highly conserved “poison” exon of EZH2 that results in nonsense-mediated decay and contributes to impaired hematopoiesis. These data provide a mechanistic basis for the enrichment of specific mutations in spliceosomal proteins in myelodysplasia, and suggest that altered RNA recognition activity is a novel mechanism of leukemogenesis. mRNA profiles of murine model and K562 cells expressing SRSF2 WT, mutants and knockdown of SRSF2 in TF-1 cells generated by deep sequencing.
Project description:Snail is a zinc-finger transcription factor best known for its ability to down-regulate E-cadherin. Its established significance in embryology and organogenesis has been expanded to include a role in the tumor progression of a number of human cancers. In addition to E-cadherin, it has more recently been associated with the down-regulation and up-regulation of a number of other genes that affect important malignant phenotypes. After establishing the presence of up-regulated Snail in human non-small cell lung cancer specimens, we used microarrays to detail the global programme of gene expression in non-small cell lung cancer cell lines stably transduced to over-express Snail as compared to vector control cell lines. Non-small cell lung cancer cell lines (H441, H292, H1437) were stably transduced with a retroviral vector to over-express Snail. Elevated Snail and a corresponding down-regulation of E-cadherin was verified in the Snail over-expressing cell lines as compared to vector control cell lines by Western analysis. RNA extraction was performed and samples submitted to the UCLA Clinical Microarray Core for hybridization to Affymetrix arrays.
Project description:Transcriptomic profiling of human pancreatic cell lines hPNE/hTert stably over-expressing DNMTs compared to GFP-hPNE/hTert over-expressing control cells after lentiviral transduction
Project description:Snail is a zinc-finger transcription factor best known for its ability to down-regulate E-cadherin. Its established significance in embryology and organogenesis has been expanded to include a role in the tumor progression of a number of human cancers. In addition to E-cadherin, it has more recently been associated with the down-regulation and up-regulation of a number of other genes that affect important malignant phenotypes. After establishing the presence of up-regulated Snail in human non-small cell lung cancer specimens, we used microarrays to detail the global programme of gene expression in non-small cell lung cancer cell lines stably transduced to over-express Snail as compared to vector control cell lines.
Project description:Recurrent Glycogen synthase kinase-3 (GSK-3) phosphorylates multiple splicing factors, including SRSF2, and regulates the splicing of a broad range of mRNAs in human cells. Inhibition of GSK-3 disrupts splicing and promotes cell death in hematopoietic cells with heterozygous mutations in SRSF2 and not in cells with wild-type splicing factors. To characterize how GSK-3 inhibition alters the cellular proteome in SRSF2-P95H/+ cells, we have performed quantitative mass spectrometry (qMS) on K562 cells with SRSF2P95H/+ knocked into the endogenous locus (PMID 30799057) and parental K562 cells, treated with the GSK-3 inhibitor CHIR99021. We focused on the mitochondrial proteome using human proteome (UniprotKB) and mitochondrial (MitoCarta 3.0) databases for protein identification, identifying 163 mitochondrial proteins whose levels are affected by SRSF2 mutation.