PMID- 37936884
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20231109
VI  - 4
IP  - 3
DP  - 2023
TI  - Receptor activator of nuclear factor-kappa B is enriched in CD9-positive 
      extracellular vesicles released by osteoclasts.
PG  - 518-529
LID - 10.20517/evcna.2023.38 [doi]
AB  - AIM: Receptor activator of nuclear factor-kappa B (RANK)-containing extracellular 
      vesicles (EVs) bind RANK-Ligand (RANKL) on osteoblasts, and thereby 
      simultaneously inhibit bone resorption and promote bone formation. Because of 
      this, they are attractive candidates for therapeutic bone anabolic agents. 
      Previously, RANK was detected in 1 in every 36 EVs from osteoclasts by immunogold 
      electron microscopy. Here, we have sought to characterize the subpopulation of 
      EVs from osteoclasts that contains RANK in more detail. METHODS: The tetraspanins 
      CD9 and CD81 were localized in osteoclasts by immunofluorescence. EVs were 
      visualized by transmission electron microscopy. A Single Particle Interferometric 
      Reflectance Imaging Sensor (SP-IRIS) and immunoaffinity isolations examined 
      whether RANK is enriched in specific types of EVs. RESULTS: Immunofluorescence 
      showed CD9 was mostly on or near the plasma membrane of osteoclasts. In contrast, 
      CD81 was localized deeper in the osteoclast's cytosolic vesicular network. By 
      interferometry, both CD9 and CD81 positive EVs from osteoclasts were small (56-83 
      nm in diameter), consistent with electron microscopy. The CD9 and CD81 EV 
      populations were mostly distinct, and only 22% of the EVs contained both markers. 
      RANK was detected by SP-IRIS in 2%-4% of the CD9-containing EVs, but not in 
      CD81-positive EVs, from mature osteoclasts. Immunomagnetic isolation of 
      CD9-containing EVs from conditioned media of osteoclasts removed most of the 
      RANK. A trace amount of RANK was isolated with CD81. CONCLUSION: RANK was 
      enriched in a subset of the CD9-positive EVs. The current study provides the 
      first report of selective localization of RANK in subsets of EVs.
FAU - Ruan, Shaobo
AU  - Ruan S
AD  - Department of Pharmaceutics, College of Pharmacy, Gainesville, FL 32610, USA.
FAU - Rody, Wellington J Jr
AU  - Rody WJ Jr
AD  - Department of Orthodontics and Dentofacial Orthopedics, University of Pittsburgh, 
      School of Dental Medicine, Pittsburgh, PA 15261, USA.
FAU - Patel, Shivani S
AU  - Patel SS
AD  - Department of Orthodontics, University of Florida College of Dentistry, 
      Gainesville, FL 32610, USA.
FAU - Hammadi, Lina I
AU  - Hammadi LI
AD  - Department of Orthodontics, University of Florida College of Dentistry, 
      Gainesville, FL 32610, USA.
FAU - Martin, Macey L
AU  - Martin ML
AD  - Department of Orthodontics, University of Florida College of Dentistry, 
      Gainesville, FL 32610, USA.
FAU - de Faria, Lorraine P
AU  - de Faria LP
AD  - Department of Biomaterials and Oral Biology, School of Dentistry, University of 
      Sao Paulo, Sao Paulo - SP 05508-000, Brazil.
FAU - Daaboul, George
AU  - Daaboul G
AD  - Unchained Labs, Pleasanton, CA 94588, USA.
FAU - Anderson, Leif S
AU  - Anderson LS
AD  - Unchained Labs, Pleasanton, CA 94588, USA.
FAU - He, Mei
AU  - He M
AD  - Department of Pharmaceutics, College of Pharmacy, Gainesville, FL 32610, USA.
FAU - Holliday, L Shannon
AU  - Holliday LS
AD  - Department of Orthodontics, University of Florida College of Dentistry, 
      Gainesville, FL 32610, USA.
AD  - Department of Anatomy & Cell Biology, University of Florida College of Medicine, 
      Gainesville, FL 32610, USA.
LA  - eng
GR  - R03 DE027504/DE/NIDCR NIH HHS/United States
GR  - R21 DE019862/DE/NIDCR NIH HHS/United States
PT  - Journal Article
DEP - 20230906
PL  - United States
TA  - Extracell Vesicles Circ Nucl Acids
JT  - Extracellular vesicles and circulating nucleic acids
JID - 101775065
PMC - PMC10629932
MID - NIHMS1929668
OTO - NOTNLM
OT  - Exosomes
OT  - RANKL reverse signaling
OT  - Single Particle Interferometric Reflectance Imaging Sensor
OT  - bone
OT  - microvesicles
OT  - remodeling
COIS- Conflicts of interest Daaboul G and Anderson LS were employed by NanoView 
      Bioscience, Brighton MA at the time of the study.
EDAT- 2023/11/08 06:43
MHDA- 2023/11/08 06:44
PMCR- 2023/11/07
CRDT- 2023/11/08 03:55
PHST- 2023/11/08 06:44 [medline]
PHST- 2023/11/08 06:43 [pubmed]
PHST- 2023/11/08 03:55 [entrez]
PHST- 2023/11/07 00:00 [pmc-release]
AID - 10.20517/evcna.2023.38 [doi]
PST - ppublish
SO  - Extracell Vesicles Circ Nucl Acids. 2023;4(3):518-529. doi: 
      10.20517/evcna.2023.38. Epub 2023 Sep 6.