Microvesicle entry into marrow cells mediates tissue-specific changes in mRNA by direct delivery of mRNA and induction of transcription
Received 27 December 2009; received in revised form 4 January 2010; accepted 5 January 2010. published online 15 January 2010.
Objective
Microvesicles have been shown to mediate intercellular communication. Previously, we have correlated entry of murine lung-derived microvesicles into murine bone marrow cells with expression of pulmonary epithelial cell-specific messenger RNA (mRNA) in these marrow cells. The present studies establish that entry of lung-derived microvesicles into marrow cells is a prerequisite for marrow expression of pulmonary epithelial cell-derived mRNA.
Materials and Methods
Murine bone marrow cells cocultured with rat lung, but separated from them using a cell-impermeable membrane (0.4-μm pore size), were analyzed using species-specific primers (for rat or mouse).
Results
These studies revealed that surfactant B and C mRNA produced by murine marrow cells were of both rat and mouse origin. Similar results were obtained using murine lung cocultured with rat bone marrow cells or when bone marrow cells were analyzed for the presence of species-specific albumin mRNA after coculture with rat or murine liver. These studies show that microvesicles both deliver mRNA to marrow cells and mediate marrow cell transcription of tissue-specific mRNA. The latter likely underlies the longer-term stable change in genetic phenotype that has been observed. We have also observed microRNA in lung-derived microvesicles, and studies with RNase-treated microvesicles indicate that microRNA negatively modulates pulmonary epithelial cell-specific mRNA levels in cocultured marrow cells. In addition, we have also observed tissue-specific expression of brain, heart, and liver mRNA in cocultured marrow cells, suggesting that microvesicle-mediated cellular phenotype change is a universal phenomena.
Conclusion
These studies suggest that cellular systems are more phenotypically labile than previously considered.
aRhode Island Hospital, The Warren Alpert Medical School of Brown University, Division of Hematology and Oncology, Providence, RI., USA
bRhode Island Hospital, The Warren Alpert Medical School of Brown University, Division of Pulmonary, Sleep and Critical Care Medicine, Providence, RI., USA
cRhode Island Hospital, Division of Core Research Laboratories, (Transmission Electron Microscopy Facility), Providence, RI., USA
dLaboratory of Retrovirology, Division of Infectious Diseases, Department of Medicine, The Warren Alpert Medical School of Brown University, Providence, RI., USA
eRhode Island Hospital, Proteomics Core, COBRE CCRD, Providence, RI., USA
Offprint requests to: Jason M. Aliotta, M.D., Divisions of Hematology/Oncology and Pulmonary, Sleep and Critical Care Medicine, Rhode Island Hospital, 7th Floor APC Building, 593 Eddy Street, Providence, RI 02903
ABSTRACT