Microvesicle-based compositions and methods
Methods and compositions for diagnosis and/or analysis of a condition in a mammal are disclosed in which RNA from microvesicles is enriched and differentiated to so obtain a result that is indicative of the condition of tissue or organ from which the microvesicle originated. In especially preferred embodiments, the condition is a neoplastic disease of a human and can be identified and staged by differential analysis of one or more distinct RNAs, optionally together with identification and analysis of a non-RNA component of the microvesicle.
1. A method of isolating and confirming the presence of a mammalian cellular RNA in a plurality of extracellular microvesicles, the method comprising the steps of:
providing a sample that includes a plurality of mammalian cells and a plurality of extracellular microvesicles;
separating the extracellular microvesicles from the plurality of mammalian cells, and differentially isolating a sub-population of microvesicles from the extracellular microvesicles using a cell-specific or an organ-specific marker;
detecting the mammalian RNA in the extracellular microvesicles or in the sub-population of microvesicles using a step of hybridization or amplification; and
using the step of detecting the mammalian RNA to confirm the presence of the RNA, wherein the mammalian RNA is a known marker of a disease.
2. A method of detecting a mammalian cellular RNA in extracellular microvesicles, the method comprising the steps of:
isolating a plurality of extracellular microvesicles;
differentially isolating a sub-population of microvesicles from the plurality of extracellular microvesicles using a cell-specific or an organ-specific marker, and isolating RNA from the sub-population of microvesicles;
detecting the mammalian RNA in the RNA isolated from the sub-population of microvesicles using a process selected from the group consisting of hybridization and amplification, wherein the mammalian RNA is a known marker of a disease.
3. The method of any one of claim 1 or claim 2 wherein the mammalian RNA has a regulatory function in the mammalian cell.
4. The method of any one of claim 1 or claim 2 wherein the mammalian RNA has a protein encoding function in the mammalian cell.
5. The method of any one of claim 1 or claim 2 wherein the microvesicles are isolated from a biological fluid.
6. The method of any one of claim 1 or claim 2 wherein the step of amplification comprises a rtPCR or a qPCR.
7. The method of any one of claim 1 or claim 2 wherein the disease is a neoplastic disease.