Extracellular vesicle ribonucleic acid (RNA) cargo as a biomarker of hyperglycemia and type 1 diabetes
Methods and compositions are provided for detecting and/or determining a patient's risk of developing hyperglycemia or type 1 diabetes. The methods are directed to analyzing the miRNA content of extracellular vesicles recovered from said patient.
1. A method of treating a patient susceptible to hyperglycemia, said method comprising
identifying the patient as susceptible to hyperglycemia, wherein said identification step comprises
obtaining the patient's biological sample;
isolating beta cell extracellular vesicles from said sample through the use of an extracellular vesicle marker selected from the group consisting of CD9 and CD63; and
quantitating two or more miRNAs selected from the group consisting of miR-21-5p, miR-21-3p, miR-217 and miR-193-3p associated with said isolated extracellular vesicles, wherein elevated levels of two or more of said miRNAs associated with said isolated extracellular vesicles identifies a patient as being susceptible to hyperglycemia; and
treating said identified patient susceptible to hyperglycemia using standard anti-diabetes therapies.
2. The method of claim 1 wherein said elevated levels of said two or more miRNAs associated with said isolated extracellular vesicles are determined based on a comparison of the identified patient's levels of said two or more miRNAs associated with said isolated extracellular vesicles to levels of said two or more miRNAs associated with isolated extracellular vesicles in a control patient.
3. The method of claim 1 wherein said elevated levels of said two or more miRNAs associated with said isolated extracellular vesicles are determined based on a comparison of the identified patient's levels of said two or more miRNAs associated with said isolated extracellular vesicles in a first sample relative to levels of said two or more miRNAs associated with said isolated extracellular vesicles in a second sample recovered from the same patient, wherein there is a temporal spacing between the recovery of said first and second sample, and said second sample has an increase in the levels of said two or more miRNAs associated with said isolated extracellular vesicles relative to the first sample.
4. The method of claim 3 wherein the temporal spacing is selected from 1 day, 1 week, 1 month, 1 year or 2 years.
5. The method of claim 1 wherein the step of identifying the patient as susceptible to hyperglycemia comprises detecting miR-21-5p, miR-21-3p, miR-217 and miR-193-3p associated with circulating extracellular vesicles in a patient, wherein said detecting step comprises
providing a plasma or serum sample obtained from said patient;
isolating beta cell extracellular vesicles from said sample through the use of an extracellular vesicle marker selected from the group consisting of CD9 and CD63; and
quantitating the miR-21-5p, miR-21-3p, miR-217 and miR-193-3p associated with said isolated extracellular vesicles, wherein the detection of elevated levels of two or more of miR-21-5p, miR-21-3p, miR-217 and miR-193-3p associated with said isolated extracellular vesicles identifies a patient as being susceptible to hyperglycemia.
6. The method of claim 5 wherein the detection of elevated levels of three or more of miR-21-5p, miR-21-3p, miR-217 and miR-193-3p associated with said isolated extracellular vesicles identifies a patient as being susceptible to hyperglycemia.
7. The method of claim 5 wherein the patient sample is a serum sample, and the isolation step comprises immunoprecipitation of the extracellular vesicles based on the presence of the extracellular vesicle marker CD9.
8. A method of treating a patient susceptible to hyperglycemia, said method comprising
identifying the patient as susceptible to hyperglycemia, wherein said identification step comprises
obtaining the patient's biological sample;
isolating beta cell extracellular vesicles from said sample through the use of an extracellular vesicle marker selected from the group consisting of CD9 and CD63; and
quantitating two or more miRNAs selected from the group consisting of miR-21-5p, miR-21-3p, miR-217 and miR-193-3p associated with said isolated extracellular vesicles, wherein detection of elevated levels of two or more of said miRNAs associated with said isolated extracellular vesicles identifies the patient as being susceptible to hyperglycemia; and
treating said identified patient susceptible to hyperglycemia by administering insulin to said identified patients.