The interplay between SRC kinase and Caspase-8 phosphorylation on Y380 sustains the aberrant activation of NFkB signaling and promotes angiogenesis and resistance to therapy in glioblastoma
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ABSTRACT:
PROVIDER: PRJNA796357 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR17563076_1.fastq.gz | Fastqsanger.gz | |||
| SRR17563076_2.fastq.gz | Fastqsanger.gz | |||
| SRR17563077_1.fastq.gz | Fastqsanger.gz | |||
| SRR17563077_2.fastq.gz | Fastqsanger.gz | |||
| SRR17563078_1.fastq.gz | Fastqsanger.gz |
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Project description:Caspase-8 is a cysteine protease that plays an essential role in apoptosis. Consistently with its canonical proapoptotic function, cancer cells may genetically or epigenetically downregulate its expression, or may dampen its enzymatic activity by promoting the expression of FLIP protein. However, unexpectedly, Caspase-8 expression is often retained in cancer even in the absence of FLIP up regulation, suggesting that alternative mechanisms may be exploited by cancer cells to cope with Caspase-8 expression. In this regard, we reported that SRC tyrosine kinase, which is aberrantly activated in many tumors, promotes Caspase-8 phosphorylation on Tyr380 and this event impairs Caspase-8 autoprocessing and full activation. Here, we investigated the significance of Caspase-8 expression in the modulation of gene expression in glioblastoma, a brain tumor where both Caspase-8 expression and SRC activity are often aberrantly upregulated. Transcriptomic analyses identified inflammatory response as a major target of Caspase-8 expression, and in particular, the NFkB signaling as one of the most affected pathways. More importantly, we could show that SRC kinase activity promotes Caspase-8 phosphorylation on Tyr380 in this context and that this event drives the assembly of a multiprotein complex that triggers NFkB activation, thereby inducing the expression of inflammatory and pro-angiogenetic factors. Moreover Caspase-8 phosphorylation on Tyr380 sustains neoangiogenesis and resistance to radiotherapy. In summary, our work identifies a novel interplay between SRC kinase and Caspase-8 that allows cancer cells to hijack Caspase-8 to sustain tumor growth.
2022-12-29 | GSE193495 | GEO
Project description:Experimental aim: identification of SRC-2 and SRC-3-related genes in MCF-7 breast cancer cells. Experimental workflow: 1. shRNA transduction. MCF-7 cells were transduced with a mix of lentiviral particles containing five individual lentiviral pLKO.1-puro short hairpin (sh) RNA plasmids targeting different sequences on SRC-2 or SRC-3 mRNA and a pLKO.1-puro empty vector control. The puromycin selections were maintained for three weeks to obtain cells containing stably integrated shRNA. 2. Sample preparation. RNA was extracted from eight independent replicates expressing the pLKO.1-puro empty shRNA vector (shKTR), from eight replicates of SRC-2 shRNA expressing cells (shSRC-2), and from seven replicates of SRC-3 shRNA expressing cells (shSRC-3). 300 ng of total RNA from each cell sample was biotin-labelled and amplified.
2019-07-01 | E-MTAB-7018 | biostudies-arrayexpress
Project description:HDXMS comparison of NFkB (p50(39-350)/RelA(19-321) vs NFkB (p50(39-350)/RelA(19-549)
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Project description:Formation of the Death-Inducing Signalling Complex (DISC) initiates the extrinsic apoptotic signalling cascade. Caspase-8 and its regulator cFLIP control death signalling by binding to the receptor via DISC-bound FADD. By elucidating the function of Caspase-10, a close homologue of caspase-8, we unexpectedly found that caspase-10 negatively regulates caspase-8-mediated cell death signalling in the DISC. We demonstrate that caspase-10 inhibits the activation of caspase-8 independent of cFLIP. Furthermore, we show that caspase-8 does not compete with other tandem DED proteins such as cFLIP or caspase-10 in binding via FADD to the receptor as current models suggest. By utilizing caspase-8 knockout cells, we demonstrate that caspase-8 has to be placed upstream of both cFLIP and caspase-10 in the DISC. We further show that DISC formation and/or stability depends on caspase-8 but is independent from its enzymatic activity. Surprisingly, we identified caspase-10 to rewire DISC-signalling to NF-kB activation and cell survival. Our data are consistent with a model in which caspase-10 and cFLIP co-ordinately regulate caspase-8-mediated cell death signalling.
2016-04-20 | GSE75365 | GEO
Project description:The non-receptor tyrosine kinase SRC is upregulated in various human cancers and plays crucial roles in cancer progression by promoting invasion and metastasis. We show that the transforming growth factor beta (TGF-β/SMAD pathway directly upregulates SRC during the epithelial-mesenchymal transition. In human epithelial MCF10A cells, TGF-β1 treatment markedly upregulated mRNA expression of SRC. Knockout of SMAD4 suppressed upregulation of SRC by TGF-β1. ChIP-sequencing analysis revealed that SRC was transcribed from the SRC1A promoter, which interacted with SMAD2 and SMAD4, in response to TGF-β1. These findings demonstrate that a direct interaction of the activated SMAD complex with the SRC1A promoter directly upregulates SRC and suggest that TGF-β contributes to SRC upregulation in the tumor microenvironment, where TGF-β-mediated tumor progression takes place.
2023-11-20 | GSE122913 | GEO
Project description:Here we have employed quantitative mass spectrometry based phosphoproteomics and proteomics to characterise the impact of knocking-down the caspase 8 gene in glioblastoma cells. This strategy enabled the identification of the pathways and processes modulated by caspase 8.
2025-12-08 | PXD060920 | Pride
Project description:Alternative mRNA splicing generates transcriptomic diversity to direct tissue-specific functions. There is a high level of alternative splicing in the brain, particularly in development, but the master regulators are poorly understood. A key splicing event early in neuronal differentiation is the inclusion of a microexon in the SH3 domain of the ubiquitous tyrosine kinase, C-SRC, to yield the constitutively active, neural-specific N1-SRC kinase. We previously demonstrated that specific inhibition of N1-SRC in developing Xenopus embryos inhibits neurogenesis, but the targets and mode of action of N1-SRC are unknown. In the current study we screened for N1-SRC SH3 domain interactors, surprisingly finding no unique targets compared to C-SRC, but rather a subset of low affinity binders, enriched in splicing regulators. Analysis of public phosphoproteomic data revealed that SRC-dependent phosphorylation of the splicing machinery is widespread and enriched in RNA binding proteins. To investigate whether N1-SRC-dependent regulation of splicing underpins its role in neurogenesis, we undertook long and short read RNAseq analysis of N1-SRC knockdown Xenopus embryos. We observed an upregulation of splicing factor expression and aberrant splicing of splicing regulators, principally HNRNPA1 and TRA2A. The affected splice junctions in both genes were in their glycine rich C-termini and enriched in SFPQ/NONO and FUS binding sites. These RNA binding proteins are SRC substrates and suggest a mechanism by which N1-SRC knockdown leads to mis-splicing of HNRNPA1 and TRA2A. Thus, the neuronal splicing of C-SRC to generate N1-SRC regulates the alternative splicing landscape during neurogenesis.
2025-08-27 | GSE290070 | GEO
Project description:Death ligands, including FAS ligand (FASL) and tumor necrosis factor (TNF), trigger apoptosis by promoting caspase-8 dimerization and activation. Impaired FAS signaling causes unconventional lymphocytes to accumulate, resulting in lymphadenopathy. Although autoprocessing of caspase-8 is considered important for apoptosis, autoprocessing-deficient Casp8 D387A/D387A mice do not develop lymphadenopathy. We show that this is because heterodimers of caspase-8 D387A and cFLIP, besides suppressing MLKL-driven necroptosis, can also induce apoptosis. Interestingly, caspase-8 D387A elicited MLKL- and caspase-1-independent intestinal atrophy and perinatal lethality in mice lacking cFLIP. Caspase-8 D387A interacted with FADD and RIPK1 in the intestine, where there was aberrant cleavage of N4BP1 and caspase-3, plus enhanced NF-κB signaling. Eliminating FADD, the adaptor protein that promotes caspase-8 oligomerization, prevented this perinatal lethality. Collectively, our results suggest that cFLIP forms heterodimers with caspase-8 D387A to promote apoptosis in some contexts, while limiting the activity of caspase-8 D387A homodimers in others.
2025-11-10 | GSE299620 | GEO
Project description:The restoration of catalytic activity to mutant enzymes by small molecules is well-established for in vitro systems. Here we show that the protein tyrosine kinase Src R388A mutant can be rescued in live cells using the small molecule imidazole. Cellular rescue of a v-Src homolog was rapid and reversible and conferred predicted oncogenic properties. Using chemical rescue in combination with mass spectrometry, six known Src kinase substrates were confirmed, and several new protein targets identified. Chemical rescue data suggests that c-Src is active under basal conditions. Rescue of R388A c-Src also allowed contributions of Src to the MAP kinase pathway to be clarified. This chemical rescue approach is likely to be of broad utility in cell signaling. We were also interested in examining the impact of Src rescue on the kinetics of gene expression. Chronic gene expression changes in v-Src transformed colon cancer and NIH3T3 cells have been reported, but the chemical rescue method permits insights into rapid kinetic changes. We used gene microarray analysis of imidazole activated Src, 1 h after imidazole treatment of 8A7F cells as well as 6N7F control cells, a set of several genes show increases (>1.7-fold) at 1 h in the 8A7F cells and another set show decreases (>1.7-fold) at 1 h with minimal changes in 6N7F control cells. Thirteen of these genes were further analyzed using real-time RT-PCR and most of the genes tested showed similar changes using both techniques. These gene changes were not reported in cells chronically transformed with v-Src or rapidly stimulated with growth factors suggesting that rapid initiation of Src-mediated tyrosine phosphorylation may induce a specialized pattern of gene expression changes. However, these earlier experiments were done under different conditions which may also contribute to gene effects. Keywords: time course, cell type comparison
2006-03-07 | GSE4043 | GEO
Project description:Cells responding to DNA damage implement complex adaptive programs that often culminate in two distinct outcomes: apoptosis or senescence. Here, to systematically identify factors driving each response, we analyzed human IMR-90 fibroblasts exposed to increasing doses of the genotoxin etoposide. We identified SRC as a key kinase contributing to this decision fork. SRC was activated by low (50 μM) etoposide but not by high (200 μM) etoposide levels. At low DNA damage, SRC-mediated activation of p38 critically promoted cell survival and senescence, and increased pro-survival BCL2L2 levels, while SRC-mediated repression of p53 prevented a rise in pro-apoptotic PUMA levels. At high DNA damage, SRC was not active, leading to elevation of p53, inhibition of p38, and apoptosis. In mice exposed to DNA damage, pharmacologic inhibition of SRC prevented the tissue accumulation of senescent cells. Inhibiting SRC could be exploited for inducing senescent-cell apoptosis in tissues to improve health outcomes.
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