Augmentation of Muscle Blood Flow by Ultrasound Cavitation is Mediated by ATP and Purinergic Signaling

Todd T. Belcik, Brian P. Davidson, Aris Xie, Melinda D. Wu, Mrinal Yadava, Qi Yue, Sherry Liang, Chae Ryung Chon, Azzdine Ammi, Joshua Field, Leanne Harmann, William M. Chilian, Joel Linden, Jonathan Lindner

    Research output: Contribution to journalArticle

    15 Citations (Scopus)

    Abstract

    BACKGROUND—: Augmentation of tissue blood flow by therapeutic ultrasound is thought to rely on convective shear. Microbubble contrast agents that undergo ultrasound-mediated cavitation markedly amplify these effects. We hypothesized that purinergic signalling is responsible for shear-dependent increases in muscle perfusion during therapeutic cavitation. METHODS—: Unilateral exposure of the proximal hindlimb of mice (with or without ischemia produced by iliac ligation) to therapeutic ultrasound (1.3 MHz, mechanical index 1.3) was performed for ten minutes after intravenous injection of 2×10 lipid microbubbles. Microvascular perfusion was evaluated by low-power contrast ultrasound perfusion imaging. In vivo muscle ATP release and in vitro ATP release from endothelial cells or erythrocytes were assessed by a luciferin-luciferase assay. Purinergic signalling pathways were assessed by studying interventions that either (1) accelerated ATP degradation; (2) inhibited P2Y receptors, adenosine receptors, or KATP channels; or (3) inhibited downstream signalling pathways involving endothelial nitric oxide synthase (eNOS) or prostanoid production (indomethacin). Augmentation in muscle perfusion by ultrasound cavitation was assessed in a proof-of-concept clinical trial in 12 subjects with stable sickle cell disease (SCD). RESULTS—: Therapeutic ultrasound cavitation increased muscle perfusion by 7-fold in normal mice, reversed tissue ischemia for up to 24 hrs in the murine model of peripheral artery disease, and doubled muscle perfusion in patients with SCD. Augmentation in flow extended well beyond the region of ultrasound exposure. Ultrasound cavitation produced a nearly 40-fold focal and sustained increase in ATP, the source of which included both endothelial cells and erythrocytes. Inhibitory studies indicated that ATP was a critical mediator of flow augmentation that acts primarily through either P2Y receptors or through adenosine produced by ectonucleotidase activity. Combined indomethacin and inhibition of eNOS abolished the effects of therapeutic ultrasound, indicating downstream signalling through both NO and prostaglandins. CONCLUSIONS—: Therapeutic ultrasound using microbubble cavitation to increase muscle perfusion relies on shear-dependent increases in ATP which can act through a diverse portfolio of purinergic signalling pathways. These events can reverse hindlimb ischemia in mice for >24 hours, and increase muscle blood flow in patients with sickle cell disease. CLINICAL TRIAL REGISTRATION—: NCT01566890 https://clinicaltrials.gov

    Original languageEnglish (US)
    JournalCirculation
    DOIs
    StateAccepted/In press - Feb 9 2017

    Fingerprint

    Adenosine Triphosphate
    Perfusion
    Muscles
    Microbubbles
    Sickle Cell Anemia
    Ischemia
    Nitric Oxide Synthase Type III
    Hindlimb
    Indomethacin
    Prostaglandins
    Endothelial Cells
    Erythrocytes
    Clinical Trials
    Therapeutics
    KATP Channels
    Purinergic P1 Receptors
    Perfusion Imaging
    Peripheral Arterial Disease
    Therapeutic Uses
    Luciferases

    ASJC Scopus subject areas

    • Cardiology and Cardiovascular Medicine
    • Physiology (medical)

    Cite this

    Augmentation of Muscle Blood Flow by Ultrasound Cavitation is Mediated by ATP and Purinergic Signaling. / Belcik, Todd T.; Davidson, Brian P.; Xie, Aris; Wu, Melinda D.; Yadava, Mrinal; Yue, Qi; Liang, Sherry; Chon, Chae Ryung; Ammi, Azzdine; Field, Joshua; Harmann, Leanne; Chilian, William M.; Linden, Joel; Lindner, Jonathan.

    In: Circulation, 09.02.2017.

    Research output: Contribution to journalArticle

    Belcik, TT, Davidson, BP, Xie, A, Wu, MD, Yadava, M, Yue, Q, Liang, S, Chon, CR, Ammi, A, Field, J, Harmann, L, Chilian, WM, Linden, J & Lindner, J 2017, 'Augmentation of Muscle Blood Flow by Ultrasound Cavitation is Mediated by ATP and Purinergic Signaling', Circulation. https://doi.org/10.1161/CIRCULATIONAHA.116.024826
    Belcik, Todd T. ; Davidson, Brian P. ; Xie, Aris ; Wu, Melinda D. ; Yadava, Mrinal ; Yue, Qi ; Liang, Sherry ; Chon, Chae Ryung ; Ammi, Azzdine ; Field, Joshua ; Harmann, Leanne ; Chilian, William M. ; Linden, Joel ; Lindner, Jonathan. / Augmentation of Muscle Blood Flow by Ultrasound Cavitation is Mediated by ATP and Purinergic Signaling. In: Circulation. 2017.
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    abstract = "BACKGROUND—: Augmentation of tissue blood flow by therapeutic ultrasound is thought to rely on convective shear. Microbubble contrast agents that undergo ultrasound-mediated cavitation markedly amplify these effects. We hypothesized that purinergic signalling is responsible for shear-dependent increases in muscle perfusion during therapeutic cavitation. METHODS—: Unilateral exposure of the proximal hindlimb of mice (with or without ischemia produced by iliac ligation) to therapeutic ultrasound (1.3 MHz, mechanical index 1.3) was performed for ten minutes after intravenous injection of 2×10 lipid microbubbles. Microvascular perfusion was evaluated by low-power contrast ultrasound perfusion imaging. In vivo muscle ATP release and in vitro ATP release from endothelial cells or erythrocytes were assessed by a luciferin-luciferase assay. Purinergic signalling pathways were assessed by studying interventions that either (1) accelerated ATP degradation; (2) inhibited P2Y receptors, adenosine receptors, or KATP channels; or (3) inhibited downstream signalling pathways involving endothelial nitric oxide synthase (eNOS) or prostanoid production (indomethacin). Augmentation in muscle perfusion by ultrasound cavitation was assessed in a proof-of-concept clinical trial in 12 subjects with stable sickle cell disease (SCD). RESULTS—: Therapeutic ultrasound cavitation increased muscle perfusion by 7-fold in normal mice, reversed tissue ischemia for up to 24 hrs in the murine model of peripheral artery disease, and doubled muscle perfusion in patients with SCD. Augmentation in flow extended well beyond the region of ultrasound exposure. Ultrasound cavitation produced a nearly 40-fold focal and sustained increase in ATP, the source of which included both endothelial cells and erythrocytes. Inhibitory studies indicated that ATP was a critical mediator of flow augmentation that acts primarily through either P2Y receptors or through adenosine produced by ectonucleotidase activity. Combined indomethacin and inhibition of eNOS abolished the effects of therapeutic ultrasound, indicating downstream signalling through both NO and prostaglandins. CONCLUSIONS—: Therapeutic ultrasound using microbubble cavitation to increase muscle perfusion relies on shear-dependent increases in ATP which can act through a diverse portfolio of purinergic signalling pathways. These events can reverse hindlimb ischemia in mice for >24 hours, and increase muscle blood flow in patients with sickle cell disease. CLINICAL TRIAL REGISTRATION—: NCT01566890 https://clinicaltrials.gov",
    author = "Belcik, {Todd T.} and Davidson, {Brian P.} and Aris Xie and Wu, {Melinda D.} and Mrinal Yadava and Qi Yue and Sherry Liang and Chon, {Chae Ryung} and Azzdine Ammi and Joshua Field and Leanne Harmann and Chilian, {William M.} and Joel Linden and Jonathan Lindner",
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    T1 - Augmentation of Muscle Blood Flow by Ultrasound Cavitation is Mediated by ATP and Purinergic Signaling

    AU - Belcik, Todd T.

    AU - Davidson, Brian P.

    AU - Xie, Aris

    AU - Wu, Melinda D.

    AU - Yadava, Mrinal

    AU - Yue, Qi

    AU - Liang, Sherry

    AU - Chon, Chae Ryung

    AU - Ammi, Azzdine

    AU - Field, Joshua

    AU - Harmann, Leanne

    AU - Chilian, William M.

    AU - Linden, Joel

    AU - Lindner, Jonathan

    PY - 2017/2/9

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    N2 - BACKGROUND—: Augmentation of tissue blood flow by therapeutic ultrasound is thought to rely on convective shear. Microbubble contrast agents that undergo ultrasound-mediated cavitation markedly amplify these effects. We hypothesized that purinergic signalling is responsible for shear-dependent increases in muscle perfusion during therapeutic cavitation. METHODS—: Unilateral exposure of the proximal hindlimb of mice (with or without ischemia produced by iliac ligation) to therapeutic ultrasound (1.3 MHz, mechanical index 1.3) was performed for ten minutes after intravenous injection of 2×10 lipid microbubbles. Microvascular perfusion was evaluated by low-power contrast ultrasound perfusion imaging. In vivo muscle ATP release and in vitro ATP release from endothelial cells or erythrocytes were assessed by a luciferin-luciferase assay. Purinergic signalling pathways were assessed by studying interventions that either (1) accelerated ATP degradation; (2) inhibited P2Y receptors, adenosine receptors, or KATP channels; or (3) inhibited downstream signalling pathways involving endothelial nitric oxide synthase (eNOS) or prostanoid production (indomethacin). Augmentation in muscle perfusion by ultrasound cavitation was assessed in a proof-of-concept clinical trial in 12 subjects with stable sickle cell disease (SCD). RESULTS—: Therapeutic ultrasound cavitation increased muscle perfusion by 7-fold in normal mice, reversed tissue ischemia for up to 24 hrs in the murine model of peripheral artery disease, and doubled muscle perfusion in patients with SCD. Augmentation in flow extended well beyond the region of ultrasound exposure. Ultrasound cavitation produced a nearly 40-fold focal and sustained increase in ATP, the source of which included both endothelial cells and erythrocytes. Inhibitory studies indicated that ATP was a critical mediator of flow augmentation that acts primarily through either P2Y receptors or through adenosine produced by ectonucleotidase activity. Combined indomethacin and inhibition of eNOS abolished the effects of therapeutic ultrasound, indicating downstream signalling through both NO and prostaglandins. CONCLUSIONS—: Therapeutic ultrasound using microbubble cavitation to increase muscle perfusion relies on shear-dependent increases in ATP which can act through a diverse portfolio of purinergic signalling pathways. These events can reverse hindlimb ischemia in mice for >24 hours, and increase muscle blood flow in patients with sickle cell disease. CLINICAL TRIAL REGISTRATION—: NCT01566890 https://clinicaltrials.gov

    AB - BACKGROUND—: Augmentation of tissue blood flow by therapeutic ultrasound is thought to rely on convective shear. Microbubble contrast agents that undergo ultrasound-mediated cavitation markedly amplify these effects. We hypothesized that purinergic signalling is responsible for shear-dependent increases in muscle perfusion during therapeutic cavitation. METHODS—: Unilateral exposure of the proximal hindlimb of mice (with or without ischemia produced by iliac ligation) to therapeutic ultrasound (1.3 MHz, mechanical index 1.3) was performed for ten minutes after intravenous injection of 2×10 lipid microbubbles. Microvascular perfusion was evaluated by low-power contrast ultrasound perfusion imaging. In vivo muscle ATP release and in vitro ATP release from endothelial cells or erythrocytes were assessed by a luciferin-luciferase assay. Purinergic signalling pathways were assessed by studying interventions that either (1) accelerated ATP degradation; (2) inhibited P2Y receptors, adenosine receptors, or KATP channels; or (3) inhibited downstream signalling pathways involving endothelial nitric oxide synthase (eNOS) or prostanoid production (indomethacin). Augmentation in muscle perfusion by ultrasound cavitation was assessed in a proof-of-concept clinical trial in 12 subjects with stable sickle cell disease (SCD). RESULTS—: Therapeutic ultrasound cavitation increased muscle perfusion by 7-fold in normal mice, reversed tissue ischemia for up to 24 hrs in the murine model of peripheral artery disease, and doubled muscle perfusion in patients with SCD. Augmentation in flow extended well beyond the region of ultrasound exposure. Ultrasound cavitation produced a nearly 40-fold focal and sustained increase in ATP, the source of which included both endothelial cells and erythrocytes. Inhibitory studies indicated that ATP was a critical mediator of flow augmentation that acts primarily through either P2Y receptors or through adenosine produced by ectonucleotidase activity. Combined indomethacin and inhibition of eNOS abolished the effects of therapeutic ultrasound, indicating downstream signalling through both NO and prostaglandins. CONCLUSIONS—: Therapeutic ultrasound using microbubble cavitation to increase muscle perfusion relies on shear-dependent increases in ATP which can act through a diverse portfolio of purinergic signalling pathways. These events can reverse hindlimb ischemia in mice for >24 hours, and increase muscle blood flow in patients with sickle cell disease. CLINICAL TRIAL REGISTRATION—: NCT01566890 https://clinicaltrials.gov

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