Document Detail


Nucleic acids within urinary exosomes/microvesicles are potential biomarkers for renal disease.
MedLine Citation:
PMID:  20428099     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Urinary exosomes or microvesicles are being studied intensively to identify potential new biomarkers for renal disease. We sought to identify whether these microvesicles contain nucleic acids. We isolated microvesicles from human urine in the same density range as that previously described for urinary exosomes and found them to have an RNA integrity profile similar to that of kidney tissue, including 18S and 28S rRNA. This profile was better preserved in urinary microvesicles compared with whole cells isolated from urine, suggesting that microvesicles may protect RNA during urine passage. We were able to detect mRNA in the human urinary microvesicles encoding proteins from all regions of the nephron and the collecting duct. Further, to provide a proof of principle, we found that microvesicles isolated from the urine of the V-ATPase B1 subunit knockout mice lacked mRNA of this subunit while containing a normal amount of the B2 subunit and aquaporin 2. The microvesicles were found to be contaminated with extraneous DNA potentially on their surface; therefore, we developed a rapid and reliable means to isolate nucleic acids from within urine microvesicles devoid of this extraneous contamination. Our study provides an experimental strategy for the routine isolation and use of urinary microvesicles as a novel and non-invasive source of nucleic acids to further renal disease biomarker discovery.
Authors:
Kevin C Miranda; Daniel T Bond; Mary McKee; Johan Skog; Teodor G Păunescu; Nicolas Da Silva; Dennis Brown; Leileata M Russo
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2010-04-28
Journal Detail:
Title:  Kidney international     Volume:  78     ISSN:  1523-1755     ISO Abbreviation:  Kidney Int.     Publication Date:  2010 Jul 
Date Detail:
Created Date:  2010-06-30     Completed Date:  2010-12-02     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0323470     Medline TA:  Kidney Int     Country:  United States    
Other Details:
Languages:  eng     Pagination:  191-9     Citation Subset:  IM    
Affiliation:
Division of Nephrology, Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts 02114, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Biological Markers / analysis*
Exosomes / chemistry*
Humans
Kidney Diseases / diagnosis*,  urine*
Mice
Microscopy, Electron, Transmission
Nucleic Acids / analysis*
Rats
Grant Support
ID/Acronym/Agency:
DK38452/DK/NIDDK NIH HHS; DK42956/DK/NIDDK NIH HHS; DK43341/DK/NIDDK NIH HHS; DK57521/DK/NIDDK NIH HHS; DK73266/DK/NIDDK NIH HHS
Chemical
Reg. No./Substance:
0/Biological Markers; 0/Nucleic Acids

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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