RNA-Containing Microvesicles and Methods Thereof

Abstract

Contemplated compositions and methods are directed to the use of microvesicles from an optionally recombinant donor cell to impart a desirable effect to a recipient cell. In certain preferred aspects, RNA of the microvesicles is employed to achieve the desirable effect. For example, microvesicles are used in vitro to increase the number of passages of a cell growing in a medium, reduce serum and/or growth factor requirements of a cell growing in a medium, and/or delay differentiation of a cell growing in a medium. Further preferred aspects include use of the microvesicles as therapeutic agents in which RNA, a membrane protein, and/or a cytosolic protein encapsulated in or coupled to the microvesicle provide a therapeutic effect. Additionally, diagnostic methods are contemplated in which RNA of a microvesicle isolated from a mammal is associated with a condition of the mammal.

1 . In a method of testing mammalian RNA, the improvement comprising:
 obtaining, from a mammalian biological fluid, a plurality of mammalian extracellular microvesicles derived from neoplastic cells;   subjecting the mammalian extracellular microvesicles to a step of RNA isolation to obtain isolated RNA, and;   (a) amplifying a nucleic acid from the isolated RNA, to thereby determine quantity and/or presence of the nucleic acid, wherein the nucleic acid is a known marker of a neoplastic disease; or   (b) hybridizing the isolated RNA to a nucleic acid, to thereby determine quantity and/or presence of the nucleic acid, wherein the nucleic acid is a known marker of a neoplastic disease.   
     
     
         2 . The method of  claim 1  wherein said isolated. RNA functions to determine the stage of a neoplastic disease. 
     
     
         3 . The method of  claim 1  wherein the mammalian RNA has a regulatory function in the mammalian cell. 
     
     
         4 . The method of  claim 1  wherein the mammalian RNA has a protein encoding function in the mammalian cell. 
     
     
         5 . The method of  claim 1  wherein the step of amplifying comprises a step of subjecting the isolated RNA to a rtPCR or a qPCR. 
     
     
         6 . The method of  claim 1  further comprising a step differentially isolating a sub-population of microvesicles from the plurality of neoplastic extracellular microvesicles using a tissue, cell or organ-specific marker prior to the step of RNA isolation. 
     
     
         7 . The method of claim of  claim 1  wherein the neoplastic disease is breast, prostate or colon cancer. 
     
     
         8 . In a method of testing mammalian RNA, the improvement comprising;
 obtaining from a mammal a biological fluid comprising a plurality of cells and a plurality of extracellular microvesicles;   separating the extracellular microvesicles from the plurality of cells;   isolating RNA from the separated extracellular microvesicles; and   a) amplifying a nucleic acid from the isolated RNA, wherein the nucleic acid is amplified from a known marker of a neoplastic disease to thereby confirm presence or quantity of the known marker to thereby test the mammalian RNA, or   b) hybridizing the isolated RNA to a nucleic acid wherein the nucleic acid is hybridized to a known marker of a neoplastic disease to thereby confirm presence or quantity of the known marker to thereby test the mammalian RNA.   
     
     
         9 . The method of  claim 8  wherein said isolated RNA functions to determine the stage of a neoplastic disease. 
     
     
         10 . The method of  claim 8  wherein the mammalian RNA has a regulatory function in the mammalian cell. 
     
     
         11 . The method of  claim 8  wherein the mammalian RNA has a protein encoding function in the mammalian cell. 
     
     
         12 . The method of  claim 8  wherein the step of amplifying comprises a step of subjecting the isolated RNA to a rtPCR or a qPCR. 
     
     
         13 . The method of  claim 8  further comprising a step of differentially isolating a sub-population of microvesicles from the plurality of neoplastic extracellular microvesicles using a tissue, cell or organ-specific marker prior to the step of RNA isolation. 
     
     
         14 . The method of  claim 8  wherein the neoplastic disease is breast, prostate or colon cancer. 
     
     
         15 . In a method of testing a plurality of extracellular microvesicles, wherein the microvesicles are from a plurality of neoplastic mammalian cells, the improvement comprising:
 obtaining, from a mammal, a biological fluid comprising a plurality of mammalian cells and a plurality of the extracellular microvesicles;   subjecting the extracellular microvesicles to a step of RNA isolation to produce isolated RNA; and   (a) amplifying a mammalian RNA from the isolated RNA; or   (b) hybridizing a nucleic acid to isolated RNA.   
     
     
         16 . The method of  claim 15  wherein said isolated RNA functions to determine the stage of a neoplastic disease. 
     
     
         17 . The method of  claim 15  wherein the mammalian RNA has a regulatory function in the mammalian cell. 
     
     
         18 . The method of  claim 15  wherein the mammalian RNA has a protein encoding function in the mammalian cell. 
     
     
         19 . The method of  claim 15  wherein the step of amplifying comprises a step of subjecting the isolated RNA to a rtPCR or a gPCR. 
     
     
         20 . The method of  claim 15  further comprising a step of differentially isolating a sub-population of microvesicles from the plurality of neoplastic extracellular microvesicles using a tissue, cell or organ-specific marker prior to the step of RNA isolation. 
     
     
         21 . The method of  claim 16  wherein the neoplastic disease is breast, prostate or colon cancer.