Does anyone know a way to block exosome „production“/„release“ in vitro? I am thinking about siRNA ore gene knockdown of genes responsible for plasma membrane fusion of multivesicular bodies, but I can't find literature dealing with this topic. However, there should be a way of doing so.
Hi lucian , GW4869 , an inhibitor of neutral sphingomyelinase , can be used to block exosome production in cultured cells. Please follow the link to access the article.
http://www.cell.com/stem-cell-reports/pdf/S2213-6711(14)00113-1.pdf
We have been able to block their release simply by using a classic secretion blocker such as monensin. Good luck
Dear Nathan, thank you for your answer. I´m not sure whether I understood you right. I thought that monesin is an inductor of exosome release rather than an inhibitor, but I might be wrong.
However, I read that Cytochalasin D or EIPA (5-ethyl-N-isopropyl amiloride) can be used to inhibit exosome release. Have you experience in any of them? I am afraid that these substances are too unspecific and intervene with other biological process.
http://www.ncbi.nlm.nih.gov/pubmed/?term=PMC3174681
I am sorry Lucian. I meant to say Cytochalasin D in my previous post and you did cite the manuscript in which we did this work. Though I cannot speak to the specificity in our hands, we were able to show that UNLIKE monensin, cyto D was able to block all quantifiable Survivin using a Survivin ELISA. I am sure we are not seeing the whole picture as I would not think that all Survivin even was exosomal yet our ELISA work would indicate this. We may be able to one day have the sensitivity we demand but not yet. Sorry for my error. Nathan
Hi lucian , GW4869 , an inhibitor of neutral sphingomyelinase , can be used to block exosome production in cultured cells. Please follow the link to access the article.
http://www.cell.com/stem-cell-reports/pdf/S2213-6711(14)00113-1.pdf
In the following paper manumycin-A was used to block exosome-release dependent on spingomyelinase-2:
http://www.ncbi.nlm.nih.gov/pubmed/?term=U+nidirectional+transfer+of+microRNA-loaded+exosomes+from+T+cells+to+antigen-presenting+cells
I think Brefeldin inhibitor is toxic to cells beyond 24 hours, so I would not recommend it for long term inhibition. GW4869 on the other hand seemed not to affect cell viability in the paper mentioned by Shashank here. I intend to try this myself as well.
Hi,
Is there a way to inhibit exosomal packaging of molecules, specifically proteins?
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