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Extracellular Vesicles from Caveolin-Enriched Microdomains Regulate Hyaluronan-Mediated Sustained Vascular Integrity.

Mirzapoiazova T, Lennon FE, Mambetsariev B, Allen M, Riehm J, Poroyko VA, Singleton PA - Int J Cell Biol (2015)

Bottom Line: These effects were blocked by inhibiting caveolin-enriched microdomain (CEM) formation.Further, inhibiting enlargeosome release by annexin II siRNA attenuated the sustained barrier enhancing effects of HMW-HA.Taken together, these results suggest that differential release of extracellular vesicles from CEM modulate the sustained HPMVEC barrier regulation by HMW-HA and LMW-HA.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Section of Pulmonary and Critical Care, Pritzker School of Medicine, The University of Chicago, Chicago, IL, USA.

ABSTRACT
Defects in vascular integrity are an initiating factor in several disease processes. We have previously reported that high molecular weight hyaluronan (HMW-HA), a major glycosaminoglycan in the body, promotes rapid signal transduction in human pulmonary microvascular endothelial cells (HPMVEC) leading to barrier enhancement. In contrast, low molecular weight hyaluronan (LMW-HA), produced in disease states by hyaluronidases and reactive oxygen species (ROS), induces HPMVEC barrier disruption. However, the mechanism(s) of sustained barrier regulation by HA are poorly defined. Our results indicate that long-term (6-24 hours) exposure of HMW-HA induced release of a novel type of extracellular vesicle from HLMVEC called enlargeosomes (characterized by AHNAK expression) while LMW-HA long-term exposure promoted release of exosomes (characterized by CD9, CD63, and CD81 expression). These effects were blocked by inhibiting caveolin-enriched microdomain (CEM) formation. Further, inhibiting enlargeosome release by annexin II siRNA attenuated the sustained barrier enhancing effects of HMW-HA. Finally, exposure of isolated enlargeosomes to HPMVEC monolayers generated barrier enhancement while exosomes led to barrier disruption. Taken together, these results suggest that differential release of extracellular vesicles from CEM modulate the sustained HPMVEC barrier regulation by HMW-HA and LMW-HA. HMW-HA-induced specialized enlargeosomes can be a potential therapeutic strategy for diseases involving impaired vascular integrity.

No MeSH data available.


Related in: MedlinePlus

The effects of isolated LMW-HA and HMW-HA-induced EV on human EC barrier function. Panel (a): HPMVEC were plated on transendothelial electrical resistance (TER) electrodes, grown to confluence, and switched to serum-free media and 0, 0.1, 1, or 10 μg/mL isolated LMW-HA-induced exosomes were then added. Addition of isolated LMW-HA-induced exosomes to human EC monolayers promoted barrier disruption in a dose-dependent manner. Panel (b): HPMVEC were plated on transendothelial electrical resistance (TER) electrodes, grown to confluence, and switched to serum-free media and 0, 0.1, 1, or 10 μg/mL isolated HMW-HA-induced enlargeosomes were then added. Addition of isolated HMW-HA-induced enlargeosomes to human EC monolayers induces barrier enhancement in a dose-dependent manner.
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fig5: The effects of isolated LMW-HA and HMW-HA-induced EV on human EC barrier function. Panel (a): HPMVEC were plated on transendothelial electrical resistance (TER) electrodes, grown to confluence, and switched to serum-free media and 0, 0.1, 1, or 10 μg/mL isolated LMW-HA-induced exosomes were then added. Addition of isolated LMW-HA-induced exosomes to human EC monolayers promoted barrier disruption in a dose-dependent manner. Panel (b): HPMVEC were plated on transendothelial electrical resistance (TER) electrodes, grown to confluence, and switched to serum-free media and 0, 0.1, 1, or 10 μg/mL isolated HMW-HA-induced enlargeosomes were then added. Addition of isolated HMW-HA-induced enlargeosomes to human EC monolayers induces barrier enhancement in a dose-dependent manner.

Mentions: Since we observed differential expression of bioactive agents in EV in Figure 4 that could impact vascular integrity, we next determined the role(s) of isolated HA-induced EV in human EC barrier function. HPMVEC were plated on transendothelial electrical resistance (TER) electrodes, grown to confluence, and switched to serum-free media and either isolated LMW-HA-induced exosomes or isolated HMW-HA-induced enlargeosomes were then added. Figure 5(a) indicates that addition of isolated LMW-HA-induced exosomes to human EC monolayers promotes barrier disruption in a dose-dependent manner. In contrast, Figure 5(b) indicates that addition of isolated HMW-HA-induced enlargeosomes to human EC monolayers induces barrier enhancement in a dose-dependent manner.


Extracellular Vesicles from Caveolin-Enriched Microdomains Regulate Hyaluronan-Mediated Sustained Vascular Integrity.

Mirzapoiazova T, Lennon FE, Mambetsariev B, Allen M, Riehm J, Poroyko VA, Singleton PA - Int J Cell Biol (2015)

The effects of isolated LMW-HA and HMW-HA-induced EV on human EC barrier function. Panel (a): HPMVEC were plated on transendothelial electrical resistance (TER) electrodes, grown to confluence, and switched to serum-free media and 0, 0.1, 1, or 10 μg/mL isolated LMW-HA-induced exosomes were then added. Addition of isolated LMW-HA-induced exosomes to human EC monolayers promoted barrier disruption in a dose-dependent manner. Panel (b): HPMVEC were plated on transendothelial electrical resistance (TER) electrodes, grown to confluence, and switched to serum-free media and 0, 0.1, 1, or 10 μg/mL isolated HMW-HA-induced enlargeosomes were then added. Addition of isolated HMW-HA-induced enlargeosomes to human EC monolayers induces barrier enhancement in a dose-dependent manner.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4581561&req=5

fig5: The effects of isolated LMW-HA and HMW-HA-induced EV on human EC barrier function. Panel (a): HPMVEC were plated on transendothelial electrical resistance (TER) electrodes, grown to confluence, and switched to serum-free media and 0, 0.1, 1, or 10 μg/mL isolated LMW-HA-induced exosomes were then added. Addition of isolated LMW-HA-induced exosomes to human EC monolayers promoted barrier disruption in a dose-dependent manner. Panel (b): HPMVEC were plated on transendothelial electrical resistance (TER) electrodes, grown to confluence, and switched to serum-free media and 0, 0.1, 1, or 10 μg/mL isolated HMW-HA-induced enlargeosomes were then added. Addition of isolated HMW-HA-induced enlargeosomes to human EC monolayers induces barrier enhancement in a dose-dependent manner.
Mentions: Since we observed differential expression of bioactive agents in EV in Figure 4 that could impact vascular integrity, we next determined the role(s) of isolated HA-induced EV in human EC barrier function. HPMVEC were plated on transendothelial electrical resistance (TER) electrodes, grown to confluence, and switched to serum-free media and either isolated LMW-HA-induced exosomes or isolated HMW-HA-induced enlargeosomes were then added. Figure 5(a) indicates that addition of isolated LMW-HA-induced exosomes to human EC monolayers promotes barrier disruption in a dose-dependent manner. In contrast, Figure 5(b) indicates that addition of isolated HMW-HA-induced enlargeosomes to human EC monolayers induces barrier enhancement in a dose-dependent manner.

Bottom Line: These effects were blocked by inhibiting caveolin-enriched microdomain (CEM) formation.Further, inhibiting enlargeosome release by annexin II siRNA attenuated the sustained barrier enhancing effects of HMW-HA.Taken together, these results suggest that differential release of extracellular vesicles from CEM modulate the sustained HPMVEC barrier regulation by HMW-HA and LMW-HA.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Section of Pulmonary and Critical Care, Pritzker School of Medicine, The University of Chicago, Chicago, IL, USA.

ABSTRACT
Defects in vascular integrity are an initiating factor in several disease processes. We have previously reported that high molecular weight hyaluronan (HMW-HA), a major glycosaminoglycan in the body, promotes rapid signal transduction in human pulmonary microvascular endothelial cells (HPMVEC) leading to barrier enhancement. In contrast, low molecular weight hyaluronan (LMW-HA), produced in disease states by hyaluronidases and reactive oxygen species (ROS), induces HPMVEC barrier disruption. However, the mechanism(s) of sustained barrier regulation by HA are poorly defined. Our results indicate that long-term (6-24 hours) exposure of HMW-HA induced release of a novel type of extracellular vesicle from HLMVEC called enlargeosomes (characterized by AHNAK expression) while LMW-HA long-term exposure promoted release of exosomes (characterized by CD9, CD63, and CD81 expression). These effects were blocked by inhibiting caveolin-enriched microdomain (CEM) formation. Further, inhibiting enlargeosome release by annexin II siRNA attenuated the sustained barrier enhancing effects of HMW-HA. Finally, exposure of isolated enlargeosomes to HPMVEC monolayers generated barrier enhancement while exosomes led to barrier disruption. Taken together, these results suggest that differential release of extracellular vesicles from CEM modulate the sustained HPMVEC barrier regulation by HMW-HA and LMW-HA. HMW-HA-induced specialized enlargeosomes can be a potential therapeutic strategy for diseases involving impaired vascular integrity.

No MeSH data available.


Related in: MedlinePlus