Treatment and diagnosis of parkinson's disease using isolated and enriched populations of biofluid-derived extracellular vesicles
The invention relates generally to a process for isolating subpopulations of EVs to identify biomarkers useful identifying, determining the progression of, and/or prognosing a disease, including a neurological disease. More particularly, the present invention relates to detection technology of various exosomal biomarkers including proteins, protein modifications, sugars, RNA, DNA, lipids, and metabolites, and combinations thereof.
1. A method for treating Parkinson disease in a subject, the method comprising:
a) contacting a biological sample from the subject with a functionalized capture surface to form a complex between the functionalized capture surface and EVs comprising the at least one cell surface marker, wherein the capture surface is functionalized with a reagent that specifically binds the at least one cell surface marker, wherein the at least one surface marker is Glutamate ionotropic receptor AMPA type subunit 2 (GluR2);
b) separating the formed complex from an unbound portion of the biological sample to obtain a captured complex;
c) washing the captured complex;
d) eluting the EVs comprising the at least one surface marker from the functionalized capture surface or lysing the EVs complexed with the functionalized capture surface;
e) extracting at least one target biomarker from the eluted or lysed EVs, wherein the at least one target biomarker is parkin (PARK2) RNA;
f) determining the expression level of the at least one biomarker extracted in step (e);
g) identifying the subject as having Parkinson disease by comparing the expression level of the at least one target biomarker to a corresponding pre-defined threshold; and;
h) administering at least one therapy for Parkinson disease to the subject.
2. The method of any one of claim 1 , wherein the at least one reagent that specifically binds the at least one cell surface marker comprises an antibody, a mixture of antibodies, a vitamin, a protein, a ligand, a lectin, a peptide, an oligonucleotide, an aptamer, or any combination thereof.
3. The method of claim 1 , wherein the biological sample is selected from the group consisting of blood, plasma, serum, urine, sputum, spinal fluid, cerebrospinal fluid, pleural fluid, nipple aspirates, lymph fluid, fluid of the respiratory, intestinal, and genitourinary tracts, tear fluid, saliva, breast milk, fluid from the lymphatic system, semen, cerebrospinal fluid, intra-organ system fluid, ascitic fluid, tumor cyst fluid, amniotic fluid and combinations thereof.
4. The method of claim 3 , wherein the biological sample is plasma or serum.
5. The method of claim 1 , wherein the functionalized capture surface comprises a population of beads or a mixture of a population of beads.
6. The method of claim 5 , wherein the beads are magnetic beads.
7. The method of claim 1 , wherein the functionalized capture surface comprises one or more of a membrane, chip, nanoparticle or population of nanoparticles, nanotube or population of nanotubes, slide, chromatography medium, or any combination thereof.
8. The method of claim 1 , wherein determining the expression level of the at least one target biomarker comprises using a transcriptomic assay, quantitative PCR, a nanostring assay, a microarray or any combination thereof.