Project description:The goal of this study is to compare the miRNA profiles of melanoma cells B16-derived exosomes with B16 cells and normal cells JB6-derived exosomes.
Project description:We wanted to correlate the protein cargo of secreted exosomes with gene expression pattern in B16-F1 and B16-F1R2. For that purpose, we performed RNA sequencing analysis of B16-F1, B16-F1R2 and B16-F1R2L (Fig.1E). We identified >3000 genes significantly up-regulated and >1000 significantly down-regulated in B16-F1R2 model compared to B16-F1, using a false discovery rate (FDR) of 0.05.
Project description:Tumor-derived exosomes are emerging as mediators of tumorigenesis with a tissue-specific address and message. We explored the function of melanoma-derived exosomes in formation of primary tumors and metastatic progression in both murine models and patients. Whereas exosomes from highly metastatic melanoma cells increased the metastatic behavior of primary tumor cells by educating bone marrow (BM) progenitor cells via the MET receptor, exosomes from low metastatic melanoma cells did not alter the incidence of metastases. Melanoma-derived exosomes induced vascular leakiness at pre-metastatic sites, and reprogrammed BM progenitor cells towards a pro-vasculogenic phenotype (c-Kit+Tie2+MET+). Reducing MET expression in tumor-derived exosomes diminished the pro-metastatic behavior of BM cells. Importantly, MET expression was upregulated in circulating BM progenitor cells (CD45-CD117low and CD45-CD117lowTIE2+) isolated from stage III and stage IV melanoma patients. Rab1a, Rab5b, Rab7, and Rab27a were highly expressed in melanoma and Rab27a RNA interference decreased exosome production and/or soluble angiogenic factors in melanoma cells, thereby preventing mobilization of BM progenitor cells, tumor growth and metastasis. Finally, we identified a melanoma signature in exosomes isolated from metastatic melanoma patients, comprised of TYRP2, VLA-4, Hsp70, an Hsp90 isoform and MET oncoprotein, which together with Rab proteins, appear to represent exosome-specific proteins with prognostic potential, and may provide new therapeutic targets. Identification of molecular finger associated to exosome effects in metastatic organs Microarray analysis of genes differentially expressed in the lungs 24 and 48 hours after B16-F10 exosome tail vein injection compared to control.
Project description:Human mesenchymal stem cell (MSC)-conditioned medium (CM) was previously reported to affect the biology of tumor cells; however, the precise mechanisms remain unclear. Here, we show that MSCs secreted 40- to 100-nm particles, which have the typical characteristics of exosomes, and these MSC-derived exosomes promoted migration of the breast cancer cell line MCF7. To further investigate the effect of MSC-exosomes on MCF7, we analyzed the gene expression profiles of MCF7 treated with or without MSC-exosomes for 24 h. Investigation of whole genome gene expression level changes in breast cancer cell line MCF7 which were treated with or without mesenchymal stem cell-derived exosomes. This study uses total RNA recovered from two samples. One sample is MCF7 treated with PBS for 24 hours and another one is MCF7 treated with mesenchymal stem cell-derived exosomes for 24hours. The ultimate concentration of mesenchymal stem cell-derived exosomes used in this experiment was 400ng/ul.
Project description:Exosomes are small RNA and protein containing vesicles that can mediate hetero- and homotypic intercellular communication between normal and malignant cells. Especially, tumor-derived exosomes are believed to mediate reprogramming of the tumor-associated stroma to favor tumor growth and metastasis. In this study we isolated exosomes from three different Ewing’s sarcoma (ES) cell lines by ultracentrifugation. Microarray analysis of ES-derived exosomes and their parental cells was performed to gain insight into the spectrum of transcripts they contain and the functions in which these transcripts might be involved in. In total we analyzed six different samples consisting of three pairs of exosomal and cellular RNA of different Ewing's sarcoma cell lines.
Project description:The raw files of the proteomics dataset are N=18 and a brief description of the files is shown below:
AF1 raw: exosomes derived from Amniotic Fluid-Mesenchymal Stromal cells 1
AF2.raw: exosomes derived from Amniotic Fluid-Mesenchymal Stromal cells 2
AFA1.raw: exosomes derived from Amniotic Fluid-Mesenchymal Stromal cells 3
AFA2.raw: exosomes derived from Amniotic Fluid-Mesenchymal Stromal cells 4
AFB1.raw: exosomes derived from Amniotic Fluid-Mesenchymal Stromal cells 5
AFB2.raw: exosomes derived from Amniotic Fluid-Mesenchymal Stromal cells 6
HL1A.raw: exosomes derived from Hepatocyte like cells 1
HL1B.raw: exosomes derived from Hepatocyte like cells 2
HL2A.raw: exosomes derived from Hepatocyte like cells 3
HL2B.raw: exosomes derived from Hepatocyte like cells 4
HL3A.raw: exosomes derived from Hepatocyte like cells 5
HL3B.raw: exosomes derived from Hepatocyte like cells 6
HPLA1.raw: exosomes derived from Hepatic Progenitor-like cells 1
HPLA2.raw: exosomes derived from Hepatic Progenitor-like cells 2
HPLB1.raw: exosomes derived from Hepatic Progenitor-like cells 3
HPLB2.raw: exosomes derived from Hepatic Progenitor-like cells 4
HPLA.raw: exosomes derived from Hepatic Progenitor-like cells 5
HPLB.raw: exosomes derived from Hepatic Progenitor-like cells 6
Raw files analysis was performed with Proteome Discoverer 1.4 (Thermo) software package, using the Sequest search engine and the Uniprot human (Homo sapiens) reviewed database, downloaded on December 15, 2017, including 20,243 entries. The search was performed using carbamidomethylation of cysteine as static and oxidation of methionine as dynamic modifications. Two missed cleavage sites, a precursor mass tolerance of 10 ppm and fragment mass tolerance of 0.05 Da were allowed. False discovery rate (FDR) validation was based on q value: target FDR : 0.05. Label free quantification was performed by utilizing the precursor ion area values exported from the total ion chromatogram as defined by the Proteome Discoverer v. 1.4.0.288 (Thermo Scientific). Output files from Proteome Discoverer were processed with R programming language for statistical computing (version 4.0.3).
The following comparisons were made: AF vs HL, AF vs HPL and HL vs HPL.
The msf files are N=18 and have the same name as the raw files above.