Project description:To investigate whether exosomes from ovarian cancer cell lines could change lncRNA expression in mesothelial cells, exosomes from ovarian cancer cells (SKOV3 cells, A2780 cells) were added to MeT-5A cells, PBS treatment was set as blank control. Total RNAs of MeT-5A cells treated with exosomes were extracted, and then lncRNA sequencing was performed.
Project description:In order to investigate the molecular mechanisms underlying the further enhancement of Atorvastatin pretreated MSC-derived exosomes (MSCATV-Exo) in cardiac protection, we performed lncRNA sequencing on exosomes secreted from ATV pretreated MSCs and non treated MSCs to identify differentially expressed lncRNA. We found that 450 lncRNAs were identified to be upregulated and 1332 lncRNAs downregulated (over 1.5 fold change) in MSCATV-Exo compared to MSC-Exo.
Project description:Peritoneal dialysis (PD) is the worldwide recognized preferred dialysis treatment for children affected by end-stage kidney disease (ESKD). However, due to the unphysiological composition of PD fluids, the peritoneal membrane (PM) of these patients may undergo structural and functional alterations, which may cause fibrosis. Several factors may accelerate this process and primary kidney disease may have a causative role. In particular, patients affected by corticoresistant primary focal segmental glomerulosclerosis), a rare glomerular disease leading to nephrotic syndrome and ESKD, seem more prone to develop peritoneal fibrosis. The mechanism causing this predisposition is still unrecognized. To better define this condition, we carried out, for the first time, a new comprehensive comparative proteomic mass spectrometry analysis of mesothelial exosomes from peritoneal dialysis effluent (PDE) of 6 pediatric patients with focal segmental glomerular sclerosis (FSGS) versus 6 patients affected by other primary renal diseases (No FSGS). Our omic study demonstrated that, despite the high overlap in the protein milieu between the two study groups, machine learning allowed a complete distinction of the whole proteomic exosome mesothelial content of FSGS versus No FSGS (with 100% accuracy). Out of the 2490 identified proteins, 40% (995) were involved in epithelial-mesenchymal transition (EMT)/fibrosis and in the transforming growth factor-β pathway. Additionally, the Weight Gene Co-expression Network Analysis algorithm identified that some of the discriminative proteins (TIMP1, CTHRC1, SPARC, CHMP4B, COL5A2, ANXA13, FNC2 and CENP-E) were also highly correlated to PD vintage, fibrosis, EMT and non-neoplastic PM disease. All together our data demonstrated that mesothelial cells of FSGS patients are more prone to activate a pro-fibrotic machinery with exosomes having a primary role in this process. Moreover, they indicated that identified FSGS-associated elements in mesothelial exosome protein could be employed as potential new biomarkers of mesothelial integrity. Finally, our results highlighted that in FSGS patients particular attention should be paid to use more biocompatible dialysis solution, reduce the length of time on PD and personalize PD regimens to minimize the risk of rapid loss of PM function or development of encapsulating peritoneal sclerosis.
Project description:To determine the mRNA expression profile of HUVEC cells treated by exosomes derived from colorectal cancer cell line HCT116 transfected with lncRNA-APC1 silenced or control vector, we performedd gene expression microArray analysis form Arraystar to examine the expression of mRNAs.
Project description:Ovarian Cancer preferentially metastasizes in association with mesothelial cell-lined surfaces.We sought to determine if mesothelial cells are required for OvCa metastasis and detect alterations in mesothelial cell gene expression upon interaction with OvCa cells. To evaluate the OvCa-induced transcriptome changes in mesothelial cells, primary human mesothelial cells from 10 patients were treated with OvCa conditioned media or ascites and RNAseq analysis was performed.
Project description:Purpose: To dissect the mechanisms of EPC-drived exosome in promoting endothelial cells reendothelialization, next-generation sequencing was used to determine exosome miRNA content and alterations in mRNA expression in HUVEC. Methods: The miRNA of EPC-drived exosomes sequencing was carried out on the HiSeq 2500 Platform (NovelBio Corp. Laboratory, Shanghai). The data analysis was carried out with the QIAseq miRNA quantification platform using unique molecular index (UMI) counts according to the manufacturer’s instructions. The mRNA analysis of three independent samples of PBS or EPC exosomes treated HUVEC was performed using the the HiSeq 2500 Platform (NovelBio Corp. Laboratory, Shanghai) following the manufacturer’s instructions. qRT–PCR validation was performed using TaqMan and SYBR Green assays. Results: The 20 most highly expressed microRNAs identified in EPC exosomes s are listed in Table 1. The genes predicted to be targeted by those microRNAs most highly expressed in EPC exosomes (top 20 microRNAs comprising ~75% of all reads, Table 1) are most significantly enriched in the functional category ‘blood vessel development’within the top 10 categories. Further, endothelial-specific miR-21-5P was the most highly expressed microRNA in EPC exosomes. Conclusions:EPC exosomes promote vascular endothelial repair by delivering functional miR-21-5p.
Project description:To further explore the underlying mechanisms of the protection functions of human milk exosmes, high throughput sequencings were used to identify differentially expressed lncRNA and mRNA profiles between human milk exosomes form term human breast milk (Term-Exos) and preterm human breast milk (Pre-Exos).
Project description:This study aimed to analyze potential biomarkers for systemic sclerosis (SSc) by constructing lncRNA–miRNA–mRNA networks in circulating exosomes (cirexos).