TY - JOUR
T1 - Comprehensive Characterization of Nanosized Extracellular Vesicles from Central and Peripheral Organs
T2 - Implications for Preclinical and Clinical Applications
AU - Chand, Subhash
AU - Jo, Ala
AU - Vellichirammal, Neetha Nanoth
AU - Gowen, Austin
AU - Guda, Chittibabu
AU - Schaal, Victoria
AU - Odegaard, Katherine
AU - Lee, Hakho
AU - Pendyala, Gurudutt
AU - Yelamanchili, Sowmya V.
N1 - Funding Information:
This study is supported by NIDA grants R21DA046855 and R01DA042379 awarded to S.V.Y., R21DA049577 to G.P. and H.L. and R01DA046852 to G.P. and S.V.Y. The bioinformatics work has been partially supported by the National Institutes of Health grants to C.G. [5P20GM103427, 1P30GM127200].
Funding Information:
We thank the University of Nebraska Medical Center Mass Spectrometry and Proteomics Core Facility for its expert assistance. The UNMC Mass Spectrometry and Proteomics Core Facility is administrated through the Office of the Vice Chancellor for Research and supported by state funds from the Nebraska Research Initiative (NRI). We also thank Tom Bargar and Nicholas Conoan for their expert technical assistance for TEM.
Publisher Copyright:
Copyright © 2020 American Chemical Society.
PY - 2020/9/25
Y1 - 2020/9/25
N2 - Extracellular vesicles (EVs) are nanosized vesicles that have been garnering a lot of attention for their valuable role as potential diagnostic markers and therapeutic vehicles for a plethora of pathologies. While EV markers from biofluids such as plasma, serum, urine, cerebrospinal fluid, and in vitro cell culture-based platforms have been extensively studied, a significant knowledge gap that remains is the characterization of specific organ-derived EVs (ODE). Here, we present a standardized protocol for isolation and characterization of purified EVs isolated from brain, heart, lung, kidney, and liver from rat and postmortem human tissue. Next, using quantitative mass spectrometry-based proteomics, we characterized the respective tissue EV proteomes that identified synaptophysin, caveolin-3, solute carrier family 22 member 2, surfactant protein B, and fatty acid-binding protein 1 as potential markers for the brain, heart, kidney, lung, and liver EV, respectively. These respective tissue-specific markers were further validated using both immunoblotting and a nanoplasmonic platform single EV imaging analysis in the two species. To summarize, our study for the first time using traditional biochemical and high-precision technology platforms provides a valuable proof-of-concept approach in defining specific ODE markers, which could further be developed as potential therapeutic candidates for respective end organ-associated pathologies.
AB - Extracellular vesicles (EVs) are nanosized vesicles that have been garnering a lot of attention for their valuable role as potential diagnostic markers and therapeutic vehicles for a plethora of pathologies. While EV markers from biofluids such as plasma, serum, urine, cerebrospinal fluid, and in vitro cell culture-based platforms have been extensively studied, a significant knowledge gap that remains is the characterization of specific organ-derived EVs (ODE). Here, we present a standardized protocol for isolation and characterization of purified EVs isolated from brain, heart, lung, kidney, and liver from rat and postmortem human tissue. Next, using quantitative mass spectrometry-based proteomics, we characterized the respective tissue EV proteomes that identified synaptophysin, caveolin-3, solute carrier family 22 member 2, surfactant protein B, and fatty acid-binding protein 1 as potential markers for the brain, heart, kidney, lung, and liver EV, respectively. These respective tissue-specific markers were further validated using both immunoblotting and a nanoplasmonic platform single EV imaging analysis in the two species. To summarize, our study for the first time using traditional biochemical and high-precision technology platforms provides a valuable proof-of-concept approach in defining specific ODE markers, which could further be developed as potential therapeutic candidates for respective end organ-associated pathologies.
KW - extracellular vesicles
KW - nanoplasmonic
KW - nanovesicles
KW - organ-derived extracellular vesicles
KW - organ-specific markers
KW - quantitative proteomics
KW - single EV imaging
UR - http://www.scopus.com/inward/record.url?scp=85091004602&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85091004602&partnerID=8YFLogxK
U2 - 10.1021/acsanm.0c01654
DO - 10.1021/acsanm.0c01654
M3 - Article
C2 - 33385108
AN - SCOPUS:85091004602
VL - 3
SP - 8906
EP - 8919
JO - ACS Applied Nano Materials
JF - ACS Applied Nano Materials
SN - 2574-0970
IS - 9
ER -