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| Status |
Public on Jun 01, 2022 |
| Title |
Characterization of zebrafish melanoma-derived interstitial EVs and their ncRNA content |
| Organism |
Danio rerio |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Extracellular vesicles (EVs) are membranous particles released by all cell types. Their role as functional carriers of bioactive molecules is boosted in cancer where they can be either secreted in biological fluids or found in the intercellular space (interstitial EVs, iEVs). Here we have opti-mised a method for the isolation and characterization of zebrafish iEVs from whole melanoma tissues. Zebrafish melanoma iEVs are in the range of ~ 140 nm by nanoparticle tracking analysis (NTA) and TEM analysis. Western blot revealed enrichment for CD63 and Alix in the iEV frac-tion, but not in melanoma cell lysates. Super resolution and confocal microscopy revealed that purified zebrafish iEVs were GFP+ indicating that they integrate the oncogene, GFP-HRASV12G within their vesicular membrane. Analysis of RNA-Seq data revealed that 118 ncRNAs are differentially distributed between zebrafish melanoma and their iEVs, with only 18 of them be-ing selectively enriched in iEVs. Among these, the RNA components of RNAses MRP and P, which process ribosomal RNA precursors, mitochondrial RNAs and some mRNAs, were enriched in iEVs. We found that melanoma iEVs induce an inflammatory response when injected in larval blood stream with increase of macrophage and induction of Interferon Responsive Genes. To clarify whether MRP and P contribute to the inflammation induced by melanoma iEVs we inject-ed larvae with MRP or P RNAs and found an inflammatory response similar to that induced by melanoma iEVs. This suggests that zebrafish melanoma iEVs are a source of MRP and P RNAs that can trigger inflammation in cells of the tumor-microenvironment.
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| Overall design |
In order to identify small RNAs preferentially accumulated in extracellular vesicles derived from zebrafish melanoma, we performed RNA sequencing analysis (RNA-Seq), of small RNAs isolated from EVs and melanoma samples.
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| Contributor(s) |
Mione MC, Biagini V, Anelli V, Kerschbamer E, Baghini M, Busi F, Gurrieri E, Notarangelo M, Piazza S, Van Niel G, D'Agostino V |
| Citation(s) |
35628321 |
| Submission date |
Nov 22, 2021 |
| Last update date |
Jun 01, 2022 |
| Contact name |
Maria Caterina Mione |
| E-mail(s) |
mariacaterina.mione@unitn.it
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| Organization name |
University of Trento
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| Department |
CIBIO
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| Lab |
Experimental Cancer Biology
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| Street address |
Via Sommarive 9
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| City |
Trento |
| ZIP/Postal code |
38123 |
| Country |
Italy |
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| Platforms (1) |
| GPL18413 |
Illumina HiSeq 2500 (Danio rerio) |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA782626 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE189352_results.txi.star.ENSDARG.smRNA.kitaRAS_Mel_vs_kitaRAS_EV.ann.csv.gz |
1.0 Mb |
(ftp)(http) |
CSV |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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