We have previously demonstrated that membrane enclosed microvesicles derived from murine lung cells contain lung-specific mRNA, micro RNA and protein and that these microvesicles enter murine marrow cells and effect a change in genetic phenotype manifest by expression of the lung-specific mRNAs;surfactants A-D, aquaporins and clara cell specific protein and in function manifest by increased conversion to epithelial lung cells after transplantation into lethally irradiated mice. The observed mRNA changes appear to be due to transfer of both mRNA and of a tissue specific transcriptional protein. Murine heart, liver and brain also effect tissue specific genetic changes in target marrow cells. The present proposal is a natural extension of these now published studies. We plan to elucidate the specific target cells in marrow for microvesicle genetic changes and will also study individual lung cells for their specific capacity to induce genetic change in marrow;previous studies were with whole lung. We plan to evaluate the stability of the observed genetic changes in marrow both in vitro and in vivo. Lastly, we will extend our characterization of the potential for reprogramming and attempt to identify specific transcriptional proteins or microRNA which may mediate the observed phenotype changes. To carry out these studies we will employ fluorescent activated cell sorting of cell populations, Real-time RT-PCR, immunohistochemistry, cell culture, murine marrow transplantation, SILAC to study protein transfer, lente viral vector transduction of cells and comparative 2-D gel electrophoresis and difference electrophoresis (DIGE). These studies should establish a more precise platform for studies of microvesicles, establish the stability of microvesicle induced genetic changes and begin to unravel the mechanisms behind such changes. This work could alter our thinking toward cell biology in general and open new strategies for approaching a number of diseases characterized by tissue degeneration or damage.

Public Health Relevance

The focus of this project is to determine if vesicles shed by lung cells are able to influence the identity of bone marrow cells that consume them to the point that they behave like lung cells. Such an approach could lead to new approaches for tissue regeneration and repair for a variety of lung diseases. Alternatively, by preventing the transfer of malignant behavior from cancerous cells to non-cancerous cells, novel therapies for cancer could be developed.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL103726-04
Application #
8645701
Study Section
Gene and Drug Delivery Systems Study Section (GDD)
Program Officer
Welniak, Lisbeth A
Project Start
2011-04-01
Project End
2015-03-31
Budget Start
2014-04-01
Budget End
2015-03-31
Support Year
4
Fiscal Year
2014
Total Cost
$378,525
Indirect Cost
$133,525
Name
Rhode Island Hospital
Department
Type
DUNS #
075710996
City
Providence
State
RI
Country
United States
Zip Code
02903
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Del Tatto, Michael; Ng, Thomas; Aliotta, Jason M et al. (2011) Marrow cell genetic phenotype change induced by human lung cancer cells. Exp Hematol 39:1072-80
Gao, Jin-Song; Zhang, Yingjie; Tang, Xiaoli et al. (2011) The Evi1, microRNA-143, K-Ras axis in colon cancer. FEBS Lett 585:693-9