Regulation of endothelial cell permeability by platelet-derived extracellular vesicles

Byron Miyazawa, Alpa Trivedi, Padma Priya Togarrati, Daniel Potter, Gyulnar Baimukanova, Lindsay Vivona, Maximillian Lin, Ernesto Lopez, Rachael Callcut, Amit K. Srivastava, Lucy Z. Kornblith, Alexander T. Fields, Martin Schreiber, Charles E. Wade, John B. Holcomb, Shibani Pati

    Research output: Contribution to journalArticle

    3 Citations (Scopus)

    Abstract

    BACKGROUND Platelet (Plt)-derived extracellular vesicles (Plt-EVs) have hemostatic properties similar to Plts. In addition to hemostasis, Plts also function to stabilize the vasculature and maintain endothelial cell (EC) barrier integrity. We hypothesized that Plt-EVs would inhibit vascular EC permeability, similar to fresh Plts. To investigate this hypothesis, we used in vitro and in vivo models of vascular endothelial compromise and bleeding. METHODS In the vitro model, Plt-EVs were isolated by ultracentrifugation and characterized for Plt markers and particle size distribution. Effects of Plts and Plt-EVs on endothelial barrier function were assessed by transendothelial electrical resistance measurements and histological analysis of endothelial junction proteins. Hemostatic potential of Plt-EVs and Plts was assessed by multiple electrode Plt aggregometry. Using an in vivo model, the effects of Plts and Plt-EVs on vascular permeability and bleeding were assessed in non-obese diabetic-severe combined immunodeficient (NOD-SCID) mice by an established Miles assay of vascular permeability and a tail snip bleeding assay. RESULTS In the in vitro model, Plt-EVs displayed exosomal size distribution and expressed Plt-specific surface markers. Platelets and Plt-EVs decreased EC permeability and restored EC junctions after thrombin challenge. Multiplate aggregometry revealed that Plt-EVs enhanced thrombin receptor-activating peptide-mediated aggregation of whole blood, whereas Plts enhanced thrombin receptor-activating peptide-, arachidonic acid-, collagen-, and adenosine diphosphate-mediated aggregation. In the in vivo model, Plt-EVs are equivalent to Plts in attenuating vascular endothelial growth factor (VEGF)-A-induced vascular permeability and uncontrolled blood loss in a tail snip hemorrhage model. CONCLUSION Our study is the first to report that Plt-EVs might provide a feasible product for transfusion in trauma patients to attenuate bleeding, inhibit vascular permeability, and mitigate the endotheliopathy of trauma.

    Original languageEnglish (US)
    Pages (from-to)931-942
    Number of pages12
    JournalJournal of Trauma and Acute Care Surgery
    Volume86
    Issue number6
    DOIs
    StatePublished - Jun 1 2019

    Fingerprint

    Permeability
    Blood Platelets
    Endothelial Cells
    thrombin receptor peptide SFLLRNP
    Capillary Permeability
    Hemorrhage
    Extracellular Vesicles
    Hemostatics
    Tail
    Intercellular Junctions
    SCID Mice
    Ultracentrifugation
    Wounds and Injuries
    Hemostasis
    Electric Impedance
    Particle Size
    Arachidonic Acid
    Thrombin
    Adenosine Diphosphate
    Vascular Endothelial Growth Factor A

    Keywords

    • barrier disruption
    • hemostasis
    • trauma
    • Vascular instability

    ASJC Scopus subject areas

    • Surgery
    • Critical Care and Intensive Care Medicine

    Cite this

    Miyazawa, B., Trivedi, A., Togarrati, P. P., Potter, D., Baimukanova, G., Vivona, L., ... Pati, S. (2019). Regulation of endothelial cell permeability by platelet-derived extracellular vesicles. Journal of Trauma and Acute Care Surgery, 86(6), 931-942. https://doi.org/10.1097/TA.0000000000002230

    Regulation of endothelial cell permeability by platelet-derived extracellular vesicles. / Miyazawa, Byron; Trivedi, Alpa; Togarrati, Padma Priya; Potter, Daniel; Baimukanova, Gyulnar; Vivona, Lindsay; Lin, Maximillian; Lopez, Ernesto; Callcut, Rachael; Srivastava, Amit K.; Kornblith, Lucy Z.; Fields, Alexander T.; Schreiber, Martin; Wade, Charles E.; Holcomb, John B.; Pati, Shibani.

    In: Journal of Trauma and Acute Care Surgery, Vol. 86, No. 6, 01.06.2019, p. 931-942.

    Research output: Contribution to journalArticle

    Miyazawa, B, Trivedi, A, Togarrati, PP, Potter, D, Baimukanova, G, Vivona, L, Lin, M, Lopez, E, Callcut, R, Srivastava, AK, Kornblith, LZ, Fields, AT, Schreiber, M, Wade, CE, Holcomb, JB & Pati, S 2019, 'Regulation of endothelial cell permeability by platelet-derived extracellular vesicles', Journal of Trauma and Acute Care Surgery, vol. 86, no. 6, pp. 931-942. https://doi.org/10.1097/TA.0000000000002230
    Miyazawa, Byron ; Trivedi, Alpa ; Togarrati, Padma Priya ; Potter, Daniel ; Baimukanova, Gyulnar ; Vivona, Lindsay ; Lin, Maximillian ; Lopez, Ernesto ; Callcut, Rachael ; Srivastava, Amit K. ; Kornblith, Lucy Z. ; Fields, Alexander T. ; Schreiber, Martin ; Wade, Charles E. ; Holcomb, John B. ; Pati, Shibani. / Regulation of endothelial cell permeability by platelet-derived extracellular vesicles. In: Journal of Trauma and Acute Care Surgery. 2019 ; Vol. 86, No. 6. pp. 931-942.
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    abstract = "BACKGROUND Platelet (Plt)-derived extracellular vesicles (Plt-EVs) have hemostatic properties similar to Plts. In addition to hemostasis, Plts also function to stabilize the vasculature and maintain endothelial cell (EC) barrier integrity. We hypothesized that Plt-EVs would inhibit vascular EC permeability, similar to fresh Plts. To investigate this hypothesis, we used in vitro and in vivo models of vascular endothelial compromise and bleeding. METHODS In the vitro model, Plt-EVs were isolated by ultracentrifugation and characterized for Plt markers and particle size distribution. Effects of Plts and Plt-EVs on endothelial barrier function were assessed by transendothelial electrical resistance measurements and histological analysis of endothelial junction proteins. Hemostatic potential of Plt-EVs and Plts was assessed by multiple electrode Plt aggregometry. Using an in vivo model, the effects of Plts and Plt-EVs on vascular permeability and bleeding were assessed in non-obese diabetic-severe combined immunodeficient (NOD-SCID) mice by an established Miles assay of vascular permeability and a tail snip bleeding assay. RESULTS In the in vitro model, Plt-EVs displayed exosomal size distribution and expressed Plt-specific surface markers. Platelets and Plt-EVs decreased EC permeability and restored EC junctions after thrombin challenge. Multiplate aggregometry revealed that Plt-EVs enhanced thrombin receptor-activating peptide-mediated aggregation of whole blood, whereas Plts enhanced thrombin receptor-activating peptide-, arachidonic acid-, collagen-, and adenosine diphosphate-mediated aggregation. In the in vivo model, Plt-EVs are equivalent to Plts in attenuating vascular endothelial growth factor (VEGF)-A-induced vascular permeability and uncontrolled blood loss in a tail snip hemorrhage model. CONCLUSION Our study is the first to report that Plt-EVs might provide a feasible product for transfusion in trauma patients to attenuate bleeding, inhibit vascular permeability, and mitigate the endotheliopathy of trauma.",
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    AU - Miyazawa, Byron

    AU - Trivedi, Alpa

    AU - Togarrati, Padma Priya

    AU - Potter, Daniel

    AU - Baimukanova, Gyulnar

    AU - Vivona, Lindsay

    AU - Lin, Maximillian

    AU - Lopez, Ernesto

    AU - Callcut, Rachael

    AU - Srivastava, Amit K.

    AU - Kornblith, Lucy Z.

    AU - Fields, Alexander T.

    AU - Schreiber, Martin

    AU - Wade, Charles E.

    AU - Holcomb, John B.

    AU - Pati, Shibani

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    N2 - BACKGROUND Platelet (Plt)-derived extracellular vesicles (Plt-EVs) have hemostatic properties similar to Plts. In addition to hemostasis, Plts also function to stabilize the vasculature and maintain endothelial cell (EC) barrier integrity. We hypothesized that Plt-EVs would inhibit vascular EC permeability, similar to fresh Plts. To investigate this hypothesis, we used in vitro and in vivo models of vascular endothelial compromise and bleeding. METHODS In the vitro model, Plt-EVs were isolated by ultracentrifugation and characterized for Plt markers and particle size distribution. Effects of Plts and Plt-EVs on endothelial barrier function were assessed by transendothelial electrical resistance measurements and histological analysis of endothelial junction proteins. Hemostatic potential of Plt-EVs and Plts was assessed by multiple electrode Plt aggregometry. Using an in vivo model, the effects of Plts and Plt-EVs on vascular permeability and bleeding were assessed in non-obese diabetic-severe combined immunodeficient (NOD-SCID) mice by an established Miles assay of vascular permeability and a tail snip bleeding assay. RESULTS In the in vitro model, Plt-EVs displayed exosomal size distribution and expressed Plt-specific surface markers. Platelets and Plt-EVs decreased EC permeability and restored EC junctions after thrombin challenge. Multiplate aggregometry revealed that Plt-EVs enhanced thrombin receptor-activating peptide-mediated aggregation of whole blood, whereas Plts enhanced thrombin receptor-activating peptide-, arachidonic acid-, collagen-, and adenosine diphosphate-mediated aggregation. In the in vivo model, Plt-EVs are equivalent to Plts in attenuating vascular endothelial growth factor (VEGF)-A-induced vascular permeability and uncontrolled blood loss in a tail snip hemorrhage model. CONCLUSION Our study is the first to report that Plt-EVs might provide a feasible product for transfusion in trauma patients to attenuate bleeding, inhibit vascular permeability, and mitigate the endotheliopathy of trauma.

    AB - BACKGROUND Platelet (Plt)-derived extracellular vesicles (Plt-EVs) have hemostatic properties similar to Plts. In addition to hemostasis, Plts also function to stabilize the vasculature and maintain endothelial cell (EC) barrier integrity. We hypothesized that Plt-EVs would inhibit vascular EC permeability, similar to fresh Plts. To investigate this hypothesis, we used in vitro and in vivo models of vascular endothelial compromise and bleeding. METHODS In the vitro model, Plt-EVs were isolated by ultracentrifugation and characterized for Plt markers and particle size distribution. Effects of Plts and Plt-EVs on endothelial barrier function were assessed by transendothelial electrical resistance measurements and histological analysis of endothelial junction proteins. Hemostatic potential of Plt-EVs and Plts was assessed by multiple electrode Plt aggregometry. Using an in vivo model, the effects of Plts and Plt-EVs on vascular permeability and bleeding were assessed in non-obese diabetic-severe combined immunodeficient (NOD-SCID) mice by an established Miles assay of vascular permeability and a tail snip bleeding assay. RESULTS In the in vitro model, Plt-EVs displayed exosomal size distribution and expressed Plt-specific surface markers. Platelets and Plt-EVs decreased EC permeability and restored EC junctions after thrombin challenge. Multiplate aggregometry revealed that Plt-EVs enhanced thrombin receptor-activating peptide-mediated aggregation of whole blood, whereas Plts enhanced thrombin receptor-activating peptide-, arachidonic acid-, collagen-, and adenosine diphosphate-mediated aggregation. In the in vivo model, Plt-EVs are equivalent to Plts in attenuating vascular endothelial growth factor (VEGF)-A-induced vascular permeability and uncontrolled blood loss in a tail snip hemorrhage model. CONCLUSION Our study is the first to report that Plt-EVs might provide a feasible product for transfusion in trauma patients to attenuate bleeding, inhibit vascular permeability, and mitigate the endotheliopathy of trauma.

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    KW - hemostasis

    KW - trauma

    KW - Vascular instability

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