Project description:We report the application of RNA sequencing technology for high-throughput profiling of normal cardiomyocytes-derived and hypertrophic cardiomyocytes- derived exosomes. The miRNA expression profiles were determined by high throughput miRNA sequencing, and 635 differentially expressed miRNAs were found. However, compared with the control group, 7 miRNAs were significantly differentially expressed in exosomes released from Ang II-treated cardiomyocytes. A total of 4 miRNAs were upregulated while 3 were downregulated. We used qRT-PCR to characterize relative expression levels of miR-155 and miR-212-3p to validate the data obtained through miRNA sequencing. The results obtained through qRT-PCR and miRNA sequencing were essentially identical. To elucidate the potential role of miRNAs in hypertrophic cardiomyocytes, the prediction of miRNA targets was performed and 11,637 genes were obtained. To reduce the false positives rate of target gene prediction, only the predicted targets within the 3 databases described above were further analyzed. Finally, 5,477 predicted target genes were selected for further investigation. Our results support the concept that exosomal microRNAs have emerged as important inflammatory response modulators regulating the cardiac hypertrophy.
Project description:Evidence from epidemiological and mechanistic studies suggests that a variety of cardiovascular diseases are associated with tumour development. However, which cell types in the diseased heart are involved in the promotion of tumour progression remains poorly understood. In this study, the role of exosomes from hypertrophic cardiomyocytes in tumour progression was investigated. A model of cardiac hypertrophy was generated in mice using transverse aortic constriction (TAC). Breast cancer cells were then implanted in model animals. Exosomes derived from AC16 cardiomyocytes treated with Ang II to induce hypertrophic growth were subsequently injected into nude mice in which breast cancer cells had previously been implanted. The results showed that exosomes from hypertrophic cardiomyocytes promoted breast cancer progression. Furthermore, transcriptome sequencing and mass spectrometric analysis demonstrated that miR-362-5p, S100A7, and S100A8 were upregulated in exosomes derived from Ang II-treated AC16 cells, which promoted the proliferation, invasion, and migration of breast cancer cells. A retrospective clinical study showed that the expression of miR-362-5p, S100A7, and S100A8 was increased in plasma exosomes obtained from patients with cardiac hypertrophy. Notably, the levels of the three factors were observed to be associated with the extent of inflammation in patients with myocardial hypertrophy. Hypertrophic cardiomyocytes promote breast cancer progression through exosomes, and this effect is mediated by S100A7, S100A8, and miRNA-362-5p contained in the exosomes released from these cells.
Project description:The inflammatory immune microenvironment plays an important role in the development of cardiac hypertrophy. Exosomes have emerged as the potent modulators of inflammatory responses. This study aimed to determine how exosomes derived from angiotensin II (Ang II)-induced hypertrophic cardiomyocytes (HCs) interfere with the inflammatory signal pathways in macrophages. Herein, we showed that increased exosome release was observed in HCs when compared to normal cardiomyocytes (NCs). Incubation of the murine macrophage cell line RAW264.7 in the presence of exosomes isolated from the culture media of HCs triggers the secretion of inflammatory cytokines interleukin (IL)-6 and IL-8. Cytokines release induced by HCs-derived exosomes was prevented by down-regulation of Argonaute2 (AGO2), suggesting that the non-coding RNAs were involved in exosome-induced inflammatory responses in RAW 264.7 macrophages. RNA sequencing assays further demonstrated that a total of seven microRNAs were differentially expressed between NCs-derived and HCs-derived exosomes. Importantly, miR-155 played a crucial role in the initiation of inflammation in macrophages. Further analyses demonstrated that HCs-derived exosomes induced the phosphorylation of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 via miR-155. Our results support the concept that exosomal microRNAs have emerged as important inflammatory response modulators regulating cardiac hypertrophy.
Project description:Cardiovascular diseases (CVDs) and cancer are among the most serious life-threatening diseases with very high morbidity and mortality rates. An increasing number of clinical studies have shown that cardiovascular disease is strongly associated with an increased risk of cancer and can contribute to cancer development, but the mechanisms associated with this need to be further explored in depth. Exosomes play an important role in promoting the development of cancer, and breast cancer is the most prevalent cancer among women. In the present study, we showed by in vivo experiments that exosomes from hypertrophic cardiomyocytes significantly promote breast cancer progression. In order to explore the mechanism by which hypertrophic cardiomyocytes promote the progression of breast cancer, we conducted whole transcriptome sequencing of exosomes and cells from normal cardiomyocytes and hypertrophic cardiomyocytes. Interestingly, the majority of miRNAs were downregulated in the exosomes of hypertrophic cardiomyocytes. In the downregulated miRNAs within the exosomes of hypertrophic cardiomyocytes, a consistent sequence GGCXGAGG was identified. We found that hnRNPQ regulates exosomal sorting of miR-10400-3p and several other miRNAs. Based on this, we validated the expression levels of plasma exosomal miRNAs in patients with myocardial hypertrophy using clinical data and patient samples, providing predictive markers for cancer progression in myocardial hypertrophy patients. Our findings suggest that hypertrophic cardiomyocytes regulate the expression of the RNA-binding protein hnRNPQ, which affects the levels of exosomal miRNAs and plays a key role in promoting the development of breast cancer.
Project description:Cardiovascular diseases (CVDs) and cancer are among the most serious life-threatening diseases with very high morbidity and mortality rates. An increasing number of clinical studies have shown that cardiovascular disease is strongly associated with an increased risk of cancer and can contribute to cancer development, but the mechanisms associated with this need to be further explored in depth. Exosomes play an important role in promoting the development of cancer, and breast cancer is the most prevalent cancer among women. In the present study, we showed by in vivo experiments that exosomes from hypertrophic cardiomyocytes significantly promote breast cancer progression. In order to explore the mechanism by which hypertrophic cardiomyocytes promote the progression of breast cancer, we conducted whole transcriptome sequencing of exosomes and cells from normal cardiomyocytes and hypertrophic cardiomyocytes. Interestingly, the majority of miRNAs were downregulated in the exosomes of hypertrophic cardiomyocytes. In the downregulated miRNAs within the exosomes of hypertrophic cardiomyocytes, a consistent sequence GGCXGAGG was identified. We found that hnRNPQ regulates exosomal sorting of miR-10400-3p and several other miRNAs. Based on this, we validated the expression levels of plasma exosomal miRNAs in patients with myocardial hypertrophy using clinical data and patient samples, providing predictive markers for cancer progression in myocardial hypertrophy patients. Our findings suggest that hypertrophic cardiomyocytes regulate the expression of the RNA-binding protein hnRNPQ, which affects the levels of exosomal miRNAs and plays a key role in promoting the development of breast cancer.
Project description:the expression characteristics of lncRNAs among hypertrophic cardiomyocytes induced by isoproterenol in rat ventricular myocytes from newborn Sprague-Dawley rats.
Project description:This experiment aimed to investigate whether exosomess released by IPSC-cardiomyocytes during hypoxia can positively influence cardiac electrophysiology and miRNA expression during hypoxic stress. A multielectrode array (MEA) system was used as an in vitro method of recording real-time cardiac electrophysiological activity, highlighting the biological effects facilitated by exosomes. Additionally, miRNA-sequencing was performed to compare miRNA expressions in exosome-preconditioned and non-preconditioned IPSC-cardiomyocytes, elucidating important modulators of cardiac electrophysiology.
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.
Project description:The aim of this project was to quantify the effects of different hypertrophic stimuli on synthesis of specific proteins in adult rat ventricular cardiomyocytes