URINE EXOSOME mRNAs AND METHODS OF USING SAME TO DETECT DIABETIC NEPHROPATHY

摘要

Embodiments of the invention relate generally to methods of identifying subjects likely to develop diabetes-associated damage to the nephron, or subjects in the early stages of diabetic nephropathy. In particular, several embodiments relate to quantification of diabetic nephropathy-associated markers by isolating RNA isolated from vesicles from patient urine samples. The levels of the marker in a subject can be compared to levels in a population having normal nephron function and/or used to to track progression of diabetic nephropathy in said subject over time.

主权项

1. A method for identifying a subject likely to develop or currently affected by diabetic nephropathy, comprising:
obtaining a sample of urine from a subject,
wherein said sample comprises vesicles that are associated with RNA; isolating the vesicles from said sample; lysing said vesicles to release said vesicle-associated RNA, wherein said vesicle-associated RNA comprises an RNA associated with diabetes-induced damage to the nephron,
wherein said RNA associated with diabetes-induced damage to the nephron is selected from the group consisting of PPARGC1A, SMAD1, UMOD, NRF2, SLC12A1 and CD24; quantifying said RNA associated with diabetes-induced damage to the nephron by a method comprising:
(i) contacting RNA from said sample with a reverse transcriptase to generate complementary DNA (cDNA), and(ii) contacting said cDNA with sense and antisense primers that are specific for one of PPARGC1A, SMAD1, UMOD, NRF2, SLC12A1, and CD24 and a DNA polymerase to generate amplified DNA; comparing the amount of quantified RNA associated with diabetes-induced damage to the nephron from said sample to the quantity of a corresponding RNA from individuals having normal kidney function; and identifying a subject as likely to develop or currently affected by diabetic nephropathy when there is a difference in the quantity of said RNA associated with diabetes-induced damage to the nephron between said subject and said the quantity of said RNA in individuals with normal kidney function.