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Study of transcriptional content in exosomes derived from untreated and growth factor-treated cultured cardiomyocytes (HL-1)

ABSTRACT: Introduction: Exosomes are nano-sized extracellular vesicles, released from various cells, which can stimulate or repress responses in target cells. We have recently shown that cultured cardiomyocytes release exosomes and that they, in turn, are involved in facilitating events in target cells by alteration of gene expression. We investigated whether external stimuli of the cardiomyocyte might influence the released exosome characteristics. Material and Methods: Exosomes were isolated from media collected from cultured cardiomyocyte (HL-1) cells with or without growth factor treatment (TGF-beta2 and PDGF-BB), with a series of differential centrifugations. The exosomes were characterized with dynamic light scattering (DLS) and Western blot and analysed with Illumina whole genome microarray gene expression. Results: An average size of 50-80 nm in diameter with no difference between treatment groups was found. Analysis of the mRNA content revealed 623 transcripts in the control group, 691 in the TGF-beta2-treated group and 362 in the PDGF-BB-treated group. 235 transcripts were common for all three groups. Conclusion: We conclude that there is a difference in mRNA content between exosomes derived from cultured cardiomyocytes stimulated with growth factors. We also conclude that all exosomes contain a basic package consisting of ribosomal transcripts and mRNAs coding for proteins with functions within the energy supply system.

ORGANISM(S): Mus musculus

PROVIDER: GSE40503 | GEO | 2013/06/20

SECONDARY ACCESSION(S): PRJNA174227

REPOSITORIES: GEO

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Study of transcriptional content in exosomes derived from untreated and growth factor-treated cultured cardiomyocytes (HL-1)

Project description:Introduction: Exosomes are nano-sized extracellular vesicles, released from various cells, which can stimulate or repress responses in target cells. We have recently shown that cultured cardiomyocytes release exosomes and that they, in turn, are involved in facilitating events in target cells by alteration of gene expression. We investigated whether external stimuli of the cardiomyocyte might influence the released exosome characteristics. Material and Methods: Exosomes were isolated from media collected from cultured cardiomyocyte (HL-1) cells with or without growth factor treatment (TGF-beta2 and PDGF-BB), with a series of differential centrifugations. The exosomes were characterized with dynamic light scattering (DLS) and Western blot and analysed with Illumina whole genome microarray gene expression. Results: An average size of 50-80 nm in diameter with no difference between treatment groups was found. Analysis of the mRNA content revealed 623 transcripts in the control group, 691 in the TGF-beta2-treated group and 362 in the PDGF-BB-treated group. 235 transcripts were common for all three groups. Conclusion: We conclude that there is a difference in mRNA content between exosomes derived from cultured cardiomyocytes stimulated with growth factors. We also conclude that all exosomes contain a basic package consisting of ribosomal transcripts and mRNAs coding for proteins with functions within the energy supply system. To study if the transcriptional content in exosomes derived from untreated and growth factor-treated cultured cardiomyocytes (HL-1) differ, and if so, can this difference be explained, 4 control (untreated) exosome samples, 4 TFG-beta2-treated cardiomyocyte-derived exosome samples and 4 PDGF-BB-treated cardiomyocyte-derived exosomes were studied.

2013-06-20 | E-GEOD-40503 | biostudies-arrayexpress

Growth factor stimulation of cardiomyocytes induces changes in the transcriptional contents of secreted exosomes

Project description:To study if the transcriptional content in exosomes derived from unntreated and growth factor treated cultured cardiomyocytes (HL-1) differ and if so, can this difference be explained.This is results for the cells.The exosome reults can be find at GSE40503. 4 control untreated samples, 4 TFG-beta2 cardiomyocyte treated samples and 4 PDGF BB cardiomyocyte were studied., this reults are from the corresponding cells (that released the exosomes), used for data filtering of the exsomedata. The exosome reults can be find at GSE40503.

2013-06-20 | E-GEOD-45207 | biostudies-arrayexpress

Analysis of mRNA-miRNA interaction network reveals the role of CAFs-derived exosomes in the immune regulation of oral squamous cell carcinoma [Cal27_mRNA]

Project description:To investigate the regulatory effect of exosomes released from CAFs cells on Cal-27 cells after hOMF is activated by PDGF to become CAFs, high-throughput sequencing was performed on co-cultured Cal-27 cells.

2023-05-19 | GSE222277 | GEO

qPCR gene expression profiling of 84 key genes involved in brain micro-vessel treatment

Project description:The Mouse osteogenesis RT² Profiler PCR Array profiles the expression of 84 key genes involved in brain micro-vessel treatment. In this experiment, we established an ex vivo cerebral microvascular model and treated it with PDGF-BB. There are two samples: one is the control group, and the other is the PDGF-BB treated group.

2023-11-13 | GSE243144 | GEO

Microarray expression profile and bioinformatics analysis of circular RNAs in the phenotypic switching of rat vascular smooth muscle cells: a preliminary study

Project description:In order to further study the role of circular RNA in the phenotypic transformation of vascular smooth muscle cells (VSMCs), the differential expression profile of circRNA in the phenotypic transition of VSMCs induced by platelet-derived growth factor-BB (PDGF-BB) was screened using chip technology. Vascular smooth muscle cells from rat thoracic aorta were induced with 20ng/ml PDGF-BB as the experimental group and compared with the control group. After induction for 24 hours, the differentially expressed circRNA was screened by circular RNA chip.

2021-12-17 | GSE190952 | GEO

Dose-dependent dual roles of PDGF-BB–PDGFR-? in vascular remodeling and opposing effects of anti-PDGF drug-induced metastasis

Project description:Anti-PDGF agents are routinely used as a key component in front-line therapy for the treatment of various cancers. However, molecular mechanisms underlying their impact on vascular remodeling in relation to the dose issue remain poorly understood. Here we show that in high PDGF-BB-producing tumors, anti-PDGF drugs significantly inhibited tumor growth and metastasis by preventing pericyte (PC) loss and vascular permeability. Surprisingly, the same anti-PDGF-BB drugs promoted tumor cell dissemination and metastasis in PDGF-BB-low-producing or negative tumors by ablating PCs from tumor vessels. At the molecular level, we show that the PDGFR-? signaling pathway in PCs mediated the opposing effects and persistent exposure of PCs to PDGF-BB led to marked downregulation of PDGFR-?. Inactivation of the PDGFR-? signaling system led to decreased levels of integrin ?1?1, resulted in impaired adhesion of PCs to collagen I, IV and laminin, two principal extracellular matrix components in blood vessels for interaction with these integrins. Our data suggest that tumor PDGF-BB levels may serve as an important biomarker for selection of tumor-bearing hosts for beneficial therapy and unsupervised practice of this group of drugs could potentially promote tumor invasion and metastasis. Pericytes were isolated and treated with PDGF-BB or control for 5 days.

2013-07-22 | E-GEOD-46564 | biostudies-arrayexpress

Dose-dependent dual roles of PDGF-BB–PDGFR-β in vascular remodeling and opposing effects of anti-PDGF drug-induced metastasis

Project description:Anti-PDGF agents are routinely used as a key component in front-line therapy for the treatment of various cancers. However, molecular mechanisms underlying their impact on vascular remodeling in relation to the dose issue remain poorly understood. Here we show that in high PDGF-BB-producing tumors, anti-PDGF drugs significantly inhibited tumor growth and metastasis by preventing pericyte (PC) loss and vascular permeability. Surprisingly, the same anti-PDGF-BB drugs promoted tumor cell dissemination and metastasis in PDGF-BB-low-producing or negative tumors by ablating PCs from tumor vessels. At the molecular level, we show that the PDGFR-β signaling pathway in PCs mediated the opposing effects and persistent exposure of PCs to PDGF-BB led to marked downregulation of PDGFR-β. Inactivation of the PDGFR-β signaling system led to decreased levels of integrin α1β1, resulted in impaired adhesion of PCs to collagen I, IV and laminin, two principal extracellular matrix components in blood vessels for interaction with these integrins. Our data suggest that tumor PDGF-BB levels may serve as an important biomarker for selection of tumor-bearing hosts for beneficial therapy and unsupervised practice of this group of drugs could potentially promote tumor invasion and metastasis.

2013-07-22 | GSE46564 | GEO

Next Generation Sequencing Fascilitates Quantitative Analysis of Differential Gene Expression in Human Pericytes treated with PDGF-BB

Project description:PDGF-BB:PDGFRβ signalling in brain pericytes is critical to the development, maintenance and function of a healthy blood-brain barrier (BBB). Furthermore, BBB impairment and pericyte loss in Alzheimer’s disease (AD) is well documented. We used tissue microarrays from control and AD brains to determine if PDGF-BB:PDGFRβ signalling components were altered in AD, and found that there was a reduction in vascular expression of PDGFB. We hypothesised that reduced PDGF-BB:PDGFRβ signalling in pericytes may have an impact on functions related to the BBB. We therefore tested the effects of PDGF-BB on primary human brain pericytes in vitro to define important signalling pathways and outcomes related to BBB function. Pericytes demonstrate a biphasic response to PDGF-BB, predominantly dependent on Akt and ERK. We determined that the actions of PDGF-BB are on-target at PDGFRβ, leading to internalisation and degradation of PDGFRβ. Using pharmacological inhibitors, we dissected distinct aspects of the PDGF-BB response that are controlled by ERK and Akt pathways. PDGF-BB promotes the proliferation of pericytes and protection from toxic stimuli through ERK signalling. In contrast, PDGF-BB:PDGFRβ signalling through Akt and NF-κB augments pericyte-derived inflammatory secretions. It may therefore be possible to supplement PDGF-BB or small molecule agonists to stabilise the cerebrovasculature in AD.

2021-11-30 | GSE189712 | GEO

Analysis of mRNA-miRNA interaction network reveals the role of CAFs-derived exosomes in the immune regulation of oral squamous cell carcinoma [Exo-miRNA]

Project description:To investigate the miRNA differences in the exosomes released from cells after hOMF is activated by PDGF to become CAFs, and to clarify the regulatory role.

2023-05-19 | GSE222276 | GEO

Analysis of mRNA-miRNA interaction network reveals the role of CAFs-derived exosomes in the immune regulation of oral squamous cell carcinoma [Exo-mRNA]

Project description:To investigate the mRNA differences in the exosomes released from cells after hOMF is activated by PDGF to become CAFs, and to clarify the regulatory role.

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