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 characterizing a first mammalian cellular RNA of 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 plurality of microvesicles from the plurality of mammalian cells, and detecting the first mammalian cellular RNA from the separated extracellular microvesicles using a step of hybridization or amplification, wherein the detected first mammalian cellular RNA is a known marker of a disease;
detecting a second mammalian cellular RNA from the sample using a second step of hybridization or amplification; and
correlating the detected first mammalian cellular RNA with the detected second mammalian cellular RNA to thereby characterize the first mammalian cellular RNA.
2. A method of isolating and characterizing a first mammalian cellular RNA of a plurality of extracellular microvesicles, the method comprising the steps of:
obtaining from a mammal a sample that comprises a plurality of extracellular microvesicles that include the first mammalian cellular RNA, wherein the sample further comprises a second mammalian cellular RNA;
detecting the first mammalian cellular RNA from the plurality of extracellular microvesicles using a step of hybridization or amplification, wherein the detected RNA is a known marker of a disease;
detecting the second mammalian cellular RNA from the sample using a second step of hybridization or amplification; and
correlating the detected first mammalian cellular RNA with the detected second mammalian cellular RNA to thereby characterize the first mammalian cellular RNA.
3. In a method of isolating a mammalian cellular RNA from a biological fluid, the improvement comprising:
obtaining from a mammal a biological fluid, wherein the biological fluid comprises a plurality of extracellular microvesicles;
isolating the plurality of extracellular microvesicles from the biological fluid; and
detecting the mammalian cellular RNA from the plurality of extracellular microvesicles using a step of hybridization or amplification, wherein the detected RNA is a known marker of a disease.
4. The method of claim 1 or claim 2 wherein the second mammalian cellular RNA is detected from the extracellular microvesicles using a step of hybridization or amplification.
5. The method of claim 1 or claim 2 wherein the detected first mammalian cellular RNA is a known marker of a neoplastic disease.
6. The method of claim 1 or claim 2 wherein the step of detecting the first mammalian cellular RNA comprises quantifying the first mammalian cellular RNA.
7. The method of claim 1 or claim 2 wherein the step of detecting the second mammalian cellular RNA comprises quantifying the first mammalian cellular RNA.
8. The method of claim 1 or claim 2 wherein the second mammalian cellular RNA is exclusively expressed in a single tissue.
9. The method of claim 1 or claim 2 wherein the step of correlating comprises a quantitative correlation of the first detected mammalian cellular RNA with the detected second mammalian cellular RNA.
10. The method of claim 1 or claim 2 wherein the sample is selected from the group consisting of blood, serum, urine, and a solid tissue.
11. The method of claim 3 wherein the biological fluid is blood, serum, or urine.
12. The method of claim 3 wherein the step of detecting the mammalian cellular RNA is performed using hybridization of the mammalian cellular RNA to a solid phase or by quantitative rtPCR.
13. The method of claim 3 wherein the detected mammalian cellular RNA is a known marker of a neoplastic disease.