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Anthony P. Davenport - One of the best experts on this subject based on the ideXlab platform.

  • BS49 Human embryonic stem cell derived cardiomyocytes express functional receptors for the Cardiovascular Peptide apelin
    Basic Science, 2019
    Co-Authors: Robyn Macrae, William G. Bernard, Rhoda E. Kuc, Maria Colzani, Tom Williams, Duuamene Nyimanu, Janet J. Maguire, Sanjay Sinha, Anthony P. Davenport
    Abstract:

    Introduction The apelin receptor is expressed throughout the Cardiovascular system, including in cardiomyocytes. Receptor activation by either of its endogenous Peptide ligands, apelin or Elabela, has a positive inotropic effect and promotes vasodilatation. Human embryonic stem cell (hESC)-derived cells have the potential for use in translational research to investigate cellular signalling, disease pathogenesis and potential novel treatments. Our aim was to determine if hESC-derived cardiomyocytes express apelin receptor protein and to quantify receptor density to ascertain if this cell type can be used as a phenotypic model for human diseases associated with apelin receptor mutations. Methods H9 hESCs were cultured to induce differentiation to beating cardiomyocytes. Saturation radioligand binding experiments were performed using [Glp65,Nle75,Tyr77][125I]apelin-13 and [Pyr1]apelin-13 to define non-specific binding. Bound radioactivity was counted and data analysed using iterative curve fitting programs to obtain values of receptor affinity (KD) and density (BMAX). Immunocytochemistry was carried out using anti-apelin receptor or anti-cardiac cell marker antibodies. Results Previous work demonstrated expression of the apelin receptor in hESC-derived cardiomyocytes at the gene level by qRT-PCR at similar levels to adult cells (figure 1A). Here, radioligand binding studies have confirmed receptor protein expression in beating hESC-derived cardiomyocytes. Binding was saturable and [125I]apelin-13 bound with sub-nanomolar affinity (0.12 nM) and receptor density found to be 21 fmol/mg (figure 1B), comparable to that found in human adult heart. Hill slope was close to 1 consistent with a single binding site for apelin in these cells. Furthermore, beating hESC-derived cardiomyocytes stained positive for apelin receptor, in addition to the standard cardiac markers including cardiac troponin T (figure 2). Conclusion These data importantly confirm that hESC-derived cardiomyocytes express apelin receptor protein at similar levels to adult human heart. Apelin receptor mutations have been identified in the 100,000 Genomes Bridge Project that are associated with rare diseases, including pulmonary arterial hypertension. Our ongoing experiments aim to pharmacologically characterise the apelinergic signalling pathway in the beating hESC-derived cardiomyocytes and we propose to generate hESC-derived phenotypic models by introducing selected apelin receptor mutations via CRISPR/Cas-9 gene editing. Conflict of interest None

  • [Pyr1]Apelin-13(1-12) Is a Biologically Active ACE2 Metabolite of the Endogenous Cardiovascular Peptide [Pyr1]Apelin-13.
    Frontiers in Neuroscience, 2017
    Co-Authors: Peiran Yang, Rhoda E. Kuc, Anthony P. Davenport, Aimee L. Brame, Alex Dyson, Mervyn Singer, Robert C. Glen, Joseph Cheriyan, Ian B. Wilkinson, Janet J. Maguire
    Abstract:

    Aims: Apelin is a predicted substrate for ACE2, a novel therapeutic target. Our aim was to demonstrate the endogenous presence of the putative ACE2 product [Pyr1]apelin-13(1-12) in human Cardiovascular tissues and to confirm it retains significant biological activity for the apelin receptor in vitro and in vivo. The minimum active apelin fragment was also investigated. Methods and Results: [Pyr1]apelin-13 incubated with recombinant human ACE2 resulted in de novo generation of [Pyr1]apelin-13(1-12) identified by mass spectrometry. Endogenous [Pyr1]apelin-13(1-12) was detected by immunostaining in human heart and lung localized to the endothelium. Expression was undetectable in lung from patients with pulmonary arterial hypertension. In human heart [Pyr1]apelin-13(1-12) (pKi = 8.04±0.06) and apelin-13(F13A) (pKi = 8.07±0.24) competed with [125I]apelin-13 binding with nanomolar affinity, 4-fold lower than for [Pyr1]apelin-13 (pKi = 8.83±0.06) whereas apelin-17 exhibited highest affinity (pKi = 9.63±0.17). The rank order of potency of Peptides to inhibit forskolin-stimulated cAMP was apelin-17 (pD2 = 10.31±0.28) > [Pyr1]apelin-13 (pD2 = 9.67±0.04)  apelin-13(F13A) (pD2 = 9.54±0.05) > [Pyr1]apelin-13(1-12) (pD2 = 9.30±0.06). The truncated Peptide apelin-13(R10M) retained nanomolar potency (pD2 = 8.70±0.04) but shorter fragments exhibited low micromolar potency. In a β-arrestin recruitment assay the rank order of potency was apelin-17 (pD2 = 10.26±0.09) >> [Pyr1]apelin-13 (pD2 = 8.43±0.08) > apelin-13(R10M) (pD2 = 8.26±0.17) > apelin-13(F13A) (pD2 = 7.98±0.04)  [Pyr1]apelin-13(1-12) (pD2 = 7.84±0.06) >> shorter fragments (pD2

  • pyr1 apelin 13 1 12 is a biologically active ace2 metabolite of the endogenous Cardiovascular Peptide pyr1 apelin 13
    Frontiers in Neuroscience, 2017
    Co-Authors: Peiran Yang, Rhoda E. Kuc, Aimee L. Brame, Alex Dyson, Mervyn Singer, Robert C. Glen, Joseph Cheriyan, Ian B. Wilkinson, Anthony P. Davenport
    Abstract:

    Aims: Apelin is a predicted substrate for ACE2, a novel therapeutic target. Our aim was to demonstrate the endogenous presence of the putative ACE2 product [Pyr1]apelin-13(1-12) in human Cardiovascular tissues and to confirm it retains significant biological activity for the apelin receptor in vitro and in vivo. The minimum active apelin fragment was also investigated. Methods and Results: [Pyr1]apelin-13 incubated with recombinant human ACE2 resulted in de novo generation of [Pyr1]apelin-13(1-12) identified by mass spectrometry. Endogenous [Pyr1]apelin-13(1-12) was detected by immunostaining in human heart and lung localized to the endothelium. Expression was undetectable in lung from patients with pulmonary arterial hypertension. In human heart [Pyr1]apelin-13(1-12) (pKi = 8.04±0.06) and apelin-13(F13A) (pKi = 8.07±0.24) competed with [125I]apelin-13 binding with nanomolar affinity, 4-fold lower than for [Pyr1]apelin-13 (pKi = 8.83±0.06) whereas apelin-17 exhibited highest affinity (pKi = 9.63±0.17). The rank order of potency of Peptides to inhibit forskolin-stimulated cAMP was apelin-17 (pD2 = 10.31±0.28) > [Pyr1]apelin-13 (pD2 = 9.67±0.04)  apelin-13(F13A) (pD2 = 9.54±0.05) > [Pyr1]apelin-13(1-12) (pD2 = 9.30±0.06). The truncated Peptide apelin-13(R10M) retained nanomolar potency (pD2 = 8.70±0.04) but shorter fragments exhibited low micromolar potency. In a β-arrestin recruitment assay the rank order of potency was apelin-17 (pD2 = 10.26±0.09) >> [Pyr1]apelin-13 (pD2 = 8.43±0.08) > apelin-13(R10M) (pD2 = 8.26±0.17) > apelin-13(F13A) (pD2 = 7.98±0.04)  [Pyr1]apelin-13(1-12) (pD2 = 7.84±0.06) >> shorter fragments (pD2<6). [Pyr1]apelin-13(1-12) and apelin-13(F13A) contracted human saphenous vein with similar sub-nanomolar potencies and [Pyr1]apelin-13(1-12) was a potent inotrope in paced mouse right ventricle and human atria. [Pyr1]apelin-13(1-12) elicited a dose-dependent decrease in blood pressure in anaesthetized rat and dose-dependent increase in forearm blood flow in human volunteers. Conclusions: We provide evidence that ACE2 cleaves [Pyr1]apelin-13 to [Pyr1]apelin-13(1-12) and this cleavage product is expressed in human Cardiovascular tissues. We have demonstrated biological activity of [Pyr1]apelin-13(1-12) at the human and rodent apelin receptor in vitro and in vivo. Our data show that reported enhanced ACE2 activity in Cardiovascular disease should not significantly compromise the beneficial effects of apelin based therapies for example in PAH.

Robert C. Glen - One of the best experts on this subject based on the ideXlab platform.

  • [Pyr1]Apelin-13(1-12) Is a Biologically Active ACE2 Metabolite of the Endogenous Cardiovascular Peptide [Pyr1]Apelin-13.
    Frontiers in Neuroscience, 2017
    Co-Authors: Peiran Yang, Rhoda E. Kuc, Anthony P. Davenport, Aimee L. Brame, Alex Dyson, Mervyn Singer, Robert C. Glen, Joseph Cheriyan, Ian B. Wilkinson, Janet J. Maguire
    Abstract:

    Aims: Apelin is a predicted substrate for ACE2, a novel therapeutic target. Our aim was to demonstrate the endogenous presence of the putative ACE2 product [Pyr1]apelin-13(1-12) in human Cardiovascular tissues and to confirm it retains significant biological activity for the apelin receptor in vitro and in vivo. The minimum active apelin fragment was also investigated. Methods and Results: [Pyr1]apelin-13 incubated with recombinant human ACE2 resulted in de novo generation of [Pyr1]apelin-13(1-12) identified by mass spectrometry. Endogenous [Pyr1]apelin-13(1-12) was detected by immunostaining in human heart and lung localized to the endothelium. Expression was undetectable in lung from patients with pulmonary arterial hypertension. In human heart [Pyr1]apelin-13(1-12) (pKi = 8.04±0.06) and apelin-13(F13A) (pKi = 8.07±0.24) competed with [125I]apelin-13 binding with nanomolar affinity, 4-fold lower than for [Pyr1]apelin-13 (pKi = 8.83±0.06) whereas apelin-17 exhibited highest affinity (pKi = 9.63±0.17). The rank order of potency of Peptides to inhibit forskolin-stimulated cAMP was apelin-17 (pD2 = 10.31±0.28) > [Pyr1]apelin-13 (pD2 = 9.67±0.04)  apelin-13(F13A) (pD2 = 9.54±0.05) > [Pyr1]apelin-13(1-12) (pD2 = 9.30±0.06). The truncated Peptide apelin-13(R10M) retained nanomolar potency (pD2 = 8.70±0.04) but shorter fragments exhibited low micromolar potency. In a β-arrestin recruitment assay the rank order of potency was apelin-17 (pD2 = 10.26±0.09) >> [Pyr1]apelin-13 (pD2 = 8.43±0.08) > apelin-13(R10M) (pD2 = 8.26±0.17) > apelin-13(F13A) (pD2 = 7.98±0.04)  [Pyr1]apelin-13(1-12) (pD2 = 7.84±0.06) >> shorter fragments (pD2

  • pyr1 apelin 13 1 12 is a biologically active ace2 metabolite of the endogenous Cardiovascular Peptide pyr1 apelin 13
    Frontiers in Neuroscience, 2017
    Co-Authors: Peiran Yang, Rhoda E. Kuc, Aimee L. Brame, Alex Dyson, Mervyn Singer, Robert C. Glen, Joseph Cheriyan, Ian B. Wilkinson, Anthony P. Davenport
    Abstract:

    Aims: Apelin is a predicted substrate for ACE2, a novel therapeutic target. Our aim was to demonstrate the endogenous presence of the putative ACE2 product [Pyr1]apelin-13(1-12) in human Cardiovascular tissues and to confirm it retains significant biological activity for the apelin receptor in vitro and in vivo. The minimum active apelin fragment was also investigated. Methods and Results: [Pyr1]apelin-13 incubated with recombinant human ACE2 resulted in de novo generation of [Pyr1]apelin-13(1-12) identified by mass spectrometry. Endogenous [Pyr1]apelin-13(1-12) was detected by immunostaining in human heart and lung localized to the endothelium. Expression was undetectable in lung from patients with pulmonary arterial hypertension. In human heart [Pyr1]apelin-13(1-12) (pKi = 8.04±0.06) and apelin-13(F13A) (pKi = 8.07±0.24) competed with [125I]apelin-13 binding with nanomolar affinity, 4-fold lower than for [Pyr1]apelin-13 (pKi = 8.83±0.06) whereas apelin-17 exhibited highest affinity (pKi = 9.63±0.17). The rank order of potency of Peptides to inhibit forskolin-stimulated cAMP was apelin-17 (pD2 = 10.31±0.28) > [Pyr1]apelin-13 (pD2 = 9.67±0.04)  apelin-13(F13A) (pD2 = 9.54±0.05) > [Pyr1]apelin-13(1-12) (pD2 = 9.30±0.06). The truncated Peptide apelin-13(R10M) retained nanomolar potency (pD2 = 8.70±0.04) but shorter fragments exhibited low micromolar potency. In a β-arrestin recruitment assay the rank order of potency was apelin-17 (pD2 = 10.26±0.09) >> [Pyr1]apelin-13 (pD2 = 8.43±0.08) > apelin-13(R10M) (pD2 = 8.26±0.17) > apelin-13(F13A) (pD2 = 7.98±0.04)  [Pyr1]apelin-13(1-12) (pD2 = 7.84±0.06) >> shorter fragments (pD2<6). [Pyr1]apelin-13(1-12) and apelin-13(F13A) contracted human saphenous vein with similar sub-nanomolar potencies and [Pyr1]apelin-13(1-12) was a potent inotrope in paced mouse right ventricle and human atria. [Pyr1]apelin-13(1-12) elicited a dose-dependent decrease in blood pressure in anaesthetized rat and dose-dependent increase in forearm blood flow in human volunteers. Conclusions: We provide evidence that ACE2 cleaves [Pyr1]apelin-13 to [Pyr1]apelin-13(1-12) and this cleavage product is expressed in human Cardiovascular tissues. We have demonstrated biological activity of [Pyr1]apelin-13(1-12) at the human and rodent apelin receptor in vitro and in vivo. Our data show that reported enhanced ACE2 activity in Cardiovascular disease should not significantly compromise the beneficial effects of apelin based therapies for example in PAH.

Peiran Yang - One of the best experts on this subject based on the ideXlab platform.

  • [Pyr1]Apelin-13(1-12) Is a Biologically Active ACE2 Metabolite of the Endogenous Cardiovascular Peptide [Pyr1]Apelin-13.
    Frontiers in Neuroscience, 2017
    Co-Authors: Peiran Yang, Rhoda E. Kuc, Anthony P. Davenport, Aimee L. Brame, Alex Dyson, Mervyn Singer, Robert C. Glen, Joseph Cheriyan, Ian B. Wilkinson, Janet J. Maguire
    Abstract:

    Aims: Apelin is a predicted substrate for ACE2, a novel therapeutic target. Our aim was to demonstrate the endogenous presence of the putative ACE2 product [Pyr1]apelin-13(1-12) in human Cardiovascular tissues and to confirm it retains significant biological activity for the apelin receptor in vitro and in vivo. The minimum active apelin fragment was also investigated. Methods and Results: [Pyr1]apelin-13 incubated with recombinant human ACE2 resulted in de novo generation of [Pyr1]apelin-13(1-12) identified by mass spectrometry. Endogenous [Pyr1]apelin-13(1-12) was detected by immunostaining in human heart and lung localized to the endothelium. Expression was undetectable in lung from patients with pulmonary arterial hypertension. In human heart [Pyr1]apelin-13(1-12) (pKi = 8.04±0.06) and apelin-13(F13A) (pKi = 8.07±0.24) competed with [125I]apelin-13 binding with nanomolar affinity, 4-fold lower than for [Pyr1]apelin-13 (pKi = 8.83±0.06) whereas apelin-17 exhibited highest affinity (pKi = 9.63±0.17). The rank order of potency of Peptides to inhibit forskolin-stimulated cAMP was apelin-17 (pD2 = 10.31±0.28) > [Pyr1]apelin-13 (pD2 = 9.67±0.04)  apelin-13(F13A) (pD2 = 9.54±0.05) > [Pyr1]apelin-13(1-12) (pD2 = 9.30±0.06). The truncated Peptide apelin-13(R10M) retained nanomolar potency (pD2 = 8.70±0.04) but shorter fragments exhibited low micromolar potency. In a β-arrestin recruitment assay the rank order of potency was apelin-17 (pD2 = 10.26±0.09) >> [Pyr1]apelin-13 (pD2 = 8.43±0.08) > apelin-13(R10M) (pD2 = 8.26±0.17) > apelin-13(F13A) (pD2 = 7.98±0.04)  [Pyr1]apelin-13(1-12) (pD2 = 7.84±0.06) >> shorter fragments (pD2

  • pyr1 apelin 13 1 12 is a biologically active ace2 metabolite of the endogenous Cardiovascular Peptide pyr1 apelin 13
    Frontiers in Neuroscience, 2017
    Co-Authors: Peiran Yang, Rhoda E. Kuc, Aimee L. Brame, Alex Dyson, Mervyn Singer, Robert C. Glen, Joseph Cheriyan, Ian B. Wilkinson, Anthony P. Davenport
    Abstract:

    Aims: Apelin is a predicted substrate for ACE2, a novel therapeutic target. Our aim was to demonstrate the endogenous presence of the putative ACE2 product [Pyr1]apelin-13(1-12) in human Cardiovascular tissues and to confirm it retains significant biological activity for the apelin receptor in vitro and in vivo. The minimum active apelin fragment was also investigated. Methods and Results: [Pyr1]apelin-13 incubated with recombinant human ACE2 resulted in de novo generation of [Pyr1]apelin-13(1-12) identified by mass spectrometry. Endogenous [Pyr1]apelin-13(1-12) was detected by immunostaining in human heart and lung localized to the endothelium. Expression was undetectable in lung from patients with pulmonary arterial hypertension. In human heart [Pyr1]apelin-13(1-12) (pKi = 8.04±0.06) and apelin-13(F13A) (pKi = 8.07±0.24) competed with [125I]apelin-13 binding with nanomolar affinity, 4-fold lower than for [Pyr1]apelin-13 (pKi = 8.83±0.06) whereas apelin-17 exhibited highest affinity (pKi = 9.63±0.17). The rank order of potency of Peptides to inhibit forskolin-stimulated cAMP was apelin-17 (pD2 = 10.31±0.28) > [Pyr1]apelin-13 (pD2 = 9.67±0.04)  apelin-13(F13A) (pD2 = 9.54±0.05) > [Pyr1]apelin-13(1-12) (pD2 = 9.30±0.06). The truncated Peptide apelin-13(R10M) retained nanomolar potency (pD2 = 8.70±0.04) but shorter fragments exhibited low micromolar potency. In a β-arrestin recruitment assay the rank order of potency was apelin-17 (pD2 = 10.26±0.09) >> [Pyr1]apelin-13 (pD2 = 8.43±0.08) > apelin-13(R10M) (pD2 = 8.26±0.17) > apelin-13(F13A) (pD2 = 7.98±0.04)  [Pyr1]apelin-13(1-12) (pD2 = 7.84±0.06) >> shorter fragments (pD2<6). [Pyr1]apelin-13(1-12) and apelin-13(F13A) contracted human saphenous vein with similar sub-nanomolar potencies and [Pyr1]apelin-13(1-12) was a potent inotrope in paced mouse right ventricle and human atria. [Pyr1]apelin-13(1-12) elicited a dose-dependent decrease in blood pressure in anaesthetized rat and dose-dependent increase in forearm blood flow in human volunteers. Conclusions: We provide evidence that ACE2 cleaves [Pyr1]apelin-13 to [Pyr1]apelin-13(1-12) and this cleavage product is expressed in human Cardiovascular tissues. We have demonstrated biological activity of [Pyr1]apelin-13(1-12) at the human and rodent apelin receptor in vitro and in vivo. Our data show that reported enhanced ACE2 activity in Cardiovascular disease should not significantly compromise the beneficial effects of apelin based therapies for example in PAH.

Rhoda E. Kuc - One of the best experts on this subject based on the ideXlab platform.

  • BS49 Human embryonic stem cell derived cardiomyocytes express functional receptors for the Cardiovascular Peptide apelin
    Basic Science, 2019
    Co-Authors: Robyn Macrae, William G. Bernard, Rhoda E. Kuc, Maria Colzani, Tom Williams, Duuamene Nyimanu, Janet J. Maguire, Sanjay Sinha, Anthony P. Davenport
    Abstract:

    Introduction The apelin receptor is expressed throughout the Cardiovascular system, including in cardiomyocytes. Receptor activation by either of its endogenous Peptide ligands, apelin or Elabela, has a positive inotropic effect and promotes vasodilatation. Human embryonic stem cell (hESC)-derived cells have the potential for use in translational research to investigate cellular signalling, disease pathogenesis and potential novel treatments. Our aim was to determine if hESC-derived cardiomyocytes express apelin receptor protein and to quantify receptor density to ascertain if this cell type can be used as a phenotypic model for human diseases associated with apelin receptor mutations. Methods H9 hESCs were cultured to induce differentiation to beating cardiomyocytes. Saturation radioligand binding experiments were performed using [Glp65,Nle75,Tyr77][125I]apelin-13 and [Pyr1]apelin-13 to define non-specific binding. Bound radioactivity was counted and data analysed using iterative curve fitting programs to obtain values of receptor affinity (KD) and density (BMAX). Immunocytochemistry was carried out using anti-apelin receptor or anti-cardiac cell marker antibodies. Results Previous work demonstrated expression of the apelin receptor in hESC-derived cardiomyocytes at the gene level by qRT-PCR at similar levels to adult cells (figure 1A). Here, radioligand binding studies have confirmed receptor protein expression in beating hESC-derived cardiomyocytes. Binding was saturable and [125I]apelin-13 bound with sub-nanomolar affinity (0.12 nM) and receptor density found to be 21 fmol/mg (figure 1B), comparable to that found in human adult heart. Hill slope was close to 1 consistent with a single binding site for apelin in these cells. Furthermore, beating hESC-derived cardiomyocytes stained positive for apelin receptor, in addition to the standard cardiac markers including cardiac troponin T (figure 2). Conclusion These data importantly confirm that hESC-derived cardiomyocytes express apelin receptor protein at similar levels to adult human heart. Apelin receptor mutations have been identified in the 100,000 Genomes Bridge Project that are associated with rare diseases, including pulmonary arterial hypertension. Our ongoing experiments aim to pharmacologically characterise the apelinergic signalling pathway in the beating hESC-derived cardiomyocytes and we propose to generate hESC-derived phenotypic models by introducing selected apelin receptor mutations via CRISPR/Cas-9 gene editing. Conflict of interest None

  • [Pyr1]Apelin-13(1-12) Is a Biologically Active ACE2 Metabolite of the Endogenous Cardiovascular Peptide [Pyr1]Apelin-13.
    Frontiers in Neuroscience, 2017
    Co-Authors: Peiran Yang, Rhoda E. Kuc, Anthony P. Davenport, Aimee L. Brame, Alex Dyson, Mervyn Singer, Robert C. Glen, Joseph Cheriyan, Ian B. Wilkinson, Janet J. Maguire
    Abstract:

    Aims: Apelin is a predicted substrate for ACE2, a novel therapeutic target. Our aim was to demonstrate the endogenous presence of the putative ACE2 product [Pyr1]apelin-13(1-12) in human Cardiovascular tissues and to confirm it retains significant biological activity for the apelin receptor in vitro and in vivo. The minimum active apelin fragment was also investigated. Methods and Results: [Pyr1]apelin-13 incubated with recombinant human ACE2 resulted in de novo generation of [Pyr1]apelin-13(1-12) identified by mass spectrometry. Endogenous [Pyr1]apelin-13(1-12) was detected by immunostaining in human heart and lung localized to the endothelium. Expression was undetectable in lung from patients with pulmonary arterial hypertension. In human heart [Pyr1]apelin-13(1-12) (pKi = 8.04±0.06) and apelin-13(F13A) (pKi = 8.07±0.24) competed with [125I]apelin-13 binding with nanomolar affinity, 4-fold lower than for [Pyr1]apelin-13 (pKi = 8.83±0.06) whereas apelin-17 exhibited highest affinity (pKi = 9.63±0.17). The rank order of potency of Peptides to inhibit forskolin-stimulated cAMP was apelin-17 (pD2 = 10.31±0.28) > [Pyr1]apelin-13 (pD2 = 9.67±0.04)  apelin-13(F13A) (pD2 = 9.54±0.05) > [Pyr1]apelin-13(1-12) (pD2 = 9.30±0.06). The truncated Peptide apelin-13(R10M) retained nanomolar potency (pD2 = 8.70±0.04) but shorter fragments exhibited low micromolar potency. In a β-arrestin recruitment assay the rank order of potency was apelin-17 (pD2 = 10.26±0.09) >> [Pyr1]apelin-13 (pD2 = 8.43±0.08) > apelin-13(R10M) (pD2 = 8.26±0.17) > apelin-13(F13A) (pD2 = 7.98±0.04)  [Pyr1]apelin-13(1-12) (pD2 = 7.84±0.06) >> shorter fragments (pD2

  • pyr1 apelin 13 1 12 is a biologically active ace2 metabolite of the endogenous Cardiovascular Peptide pyr1 apelin 13
    Frontiers in Neuroscience, 2017
    Co-Authors: Peiran Yang, Rhoda E. Kuc, Aimee L. Brame, Alex Dyson, Mervyn Singer, Robert C. Glen, Joseph Cheriyan, Ian B. Wilkinson, Anthony P. Davenport
    Abstract:

    Aims: Apelin is a predicted substrate for ACE2, a novel therapeutic target. Our aim was to demonstrate the endogenous presence of the putative ACE2 product [Pyr1]apelin-13(1-12) in human Cardiovascular tissues and to confirm it retains significant biological activity for the apelin receptor in vitro and in vivo. The minimum active apelin fragment was also investigated. Methods and Results: [Pyr1]apelin-13 incubated with recombinant human ACE2 resulted in de novo generation of [Pyr1]apelin-13(1-12) identified by mass spectrometry. Endogenous [Pyr1]apelin-13(1-12) was detected by immunostaining in human heart and lung localized to the endothelium. Expression was undetectable in lung from patients with pulmonary arterial hypertension. In human heart [Pyr1]apelin-13(1-12) (pKi = 8.04±0.06) and apelin-13(F13A) (pKi = 8.07±0.24) competed with [125I]apelin-13 binding with nanomolar affinity, 4-fold lower than for [Pyr1]apelin-13 (pKi = 8.83±0.06) whereas apelin-17 exhibited highest affinity (pKi = 9.63±0.17). The rank order of potency of Peptides to inhibit forskolin-stimulated cAMP was apelin-17 (pD2 = 10.31±0.28) > [Pyr1]apelin-13 (pD2 = 9.67±0.04)  apelin-13(F13A) (pD2 = 9.54±0.05) > [Pyr1]apelin-13(1-12) (pD2 = 9.30±0.06). The truncated Peptide apelin-13(R10M) retained nanomolar potency (pD2 = 8.70±0.04) but shorter fragments exhibited low micromolar potency. In a β-arrestin recruitment assay the rank order of potency was apelin-17 (pD2 = 10.26±0.09) >> [Pyr1]apelin-13 (pD2 = 8.43±0.08) > apelin-13(R10M) (pD2 = 8.26±0.17) > apelin-13(F13A) (pD2 = 7.98±0.04)  [Pyr1]apelin-13(1-12) (pD2 = 7.84±0.06) >> shorter fragments (pD2<6). [Pyr1]apelin-13(1-12) and apelin-13(F13A) contracted human saphenous vein with similar sub-nanomolar potencies and [Pyr1]apelin-13(1-12) was a potent inotrope in paced mouse right ventricle and human atria. [Pyr1]apelin-13(1-12) elicited a dose-dependent decrease in blood pressure in anaesthetized rat and dose-dependent increase in forearm blood flow in human volunteers. Conclusions: We provide evidence that ACE2 cleaves [Pyr1]apelin-13 to [Pyr1]apelin-13(1-12) and this cleavage product is expressed in human Cardiovascular tissues. We have demonstrated biological activity of [Pyr1]apelin-13(1-12) at the human and rodent apelin receptor in vitro and in vivo. Our data show that reported enhanced ACE2 activity in Cardiovascular disease should not significantly compromise the beneficial effects of apelin based therapies for example in PAH.

Aimee L. Brame - One of the best experts on this subject based on the ideXlab platform.

  • [Pyr1]Apelin-13(1-12) Is a Biologically Active ACE2 Metabolite of the Endogenous Cardiovascular Peptide [Pyr1]Apelin-13.
    Frontiers in Neuroscience, 2017
    Co-Authors: Peiran Yang, Rhoda E. Kuc, Anthony P. Davenport, Aimee L. Brame, Alex Dyson, Mervyn Singer, Robert C. Glen, Joseph Cheriyan, Ian B. Wilkinson, Janet J. Maguire
    Abstract:

    Aims: Apelin is a predicted substrate for ACE2, a novel therapeutic target. Our aim was to demonstrate the endogenous presence of the putative ACE2 product [Pyr1]apelin-13(1-12) in human Cardiovascular tissues and to confirm it retains significant biological activity for the apelin receptor in vitro and in vivo. The minimum active apelin fragment was also investigated. Methods and Results: [Pyr1]apelin-13 incubated with recombinant human ACE2 resulted in de novo generation of [Pyr1]apelin-13(1-12) identified by mass spectrometry. Endogenous [Pyr1]apelin-13(1-12) was detected by immunostaining in human heart and lung localized to the endothelium. Expression was undetectable in lung from patients with pulmonary arterial hypertension. In human heart [Pyr1]apelin-13(1-12) (pKi = 8.04±0.06) and apelin-13(F13A) (pKi = 8.07±0.24) competed with [125I]apelin-13 binding with nanomolar affinity, 4-fold lower than for [Pyr1]apelin-13 (pKi = 8.83±0.06) whereas apelin-17 exhibited highest affinity (pKi = 9.63±0.17). The rank order of potency of Peptides to inhibit forskolin-stimulated cAMP was apelin-17 (pD2 = 10.31±0.28) > [Pyr1]apelin-13 (pD2 = 9.67±0.04)  apelin-13(F13A) (pD2 = 9.54±0.05) > [Pyr1]apelin-13(1-12) (pD2 = 9.30±0.06). The truncated Peptide apelin-13(R10M) retained nanomolar potency (pD2 = 8.70±0.04) but shorter fragments exhibited low micromolar potency. In a β-arrestin recruitment assay the rank order of potency was apelin-17 (pD2 = 10.26±0.09) >> [Pyr1]apelin-13 (pD2 = 8.43±0.08) > apelin-13(R10M) (pD2 = 8.26±0.17) > apelin-13(F13A) (pD2 = 7.98±0.04)  [Pyr1]apelin-13(1-12) (pD2 = 7.84±0.06) >> shorter fragments (pD2

  • pyr1 apelin 13 1 12 is a biologically active ace2 metabolite of the endogenous Cardiovascular Peptide pyr1 apelin 13
    Frontiers in Neuroscience, 2017
    Co-Authors: Peiran Yang, Rhoda E. Kuc, Aimee L. Brame, Alex Dyson, Mervyn Singer, Robert C. Glen, Joseph Cheriyan, Ian B. Wilkinson, Anthony P. Davenport
    Abstract:

    Aims: Apelin is a predicted substrate for ACE2, a novel therapeutic target. Our aim was to demonstrate the endogenous presence of the putative ACE2 product [Pyr1]apelin-13(1-12) in human Cardiovascular tissues and to confirm it retains significant biological activity for the apelin receptor in vitro and in vivo. The minimum active apelin fragment was also investigated. Methods and Results: [Pyr1]apelin-13 incubated with recombinant human ACE2 resulted in de novo generation of [Pyr1]apelin-13(1-12) identified by mass spectrometry. Endogenous [Pyr1]apelin-13(1-12) was detected by immunostaining in human heart and lung localized to the endothelium. Expression was undetectable in lung from patients with pulmonary arterial hypertension. In human heart [Pyr1]apelin-13(1-12) (pKi = 8.04±0.06) and apelin-13(F13A) (pKi = 8.07±0.24) competed with [125I]apelin-13 binding with nanomolar affinity, 4-fold lower than for [Pyr1]apelin-13 (pKi = 8.83±0.06) whereas apelin-17 exhibited highest affinity (pKi = 9.63±0.17). The rank order of potency of Peptides to inhibit forskolin-stimulated cAMP was apelin-17 (pD2 = 10.31±0.28) > [Pyr1]apelin-13 (pD2 = 9.67±0.04)  apelin-13(F13A) (pD2 = 9.54±0.05) > [Pyr1]apelin-13(1-12) (pD2 = 9.30±0.06). The truncated Peptide apelin-13(R10M) retained nanomolar potency (pD2 = 8.70±0.04) but shorter fragments exhibited low micromolar potency. In a β-arrestin recruitment assay the rank order of potency was apelin-17 (pD2 = 10.26±0.09) >> [Pyr1]apelin-13 (pD2 = 8.43±0.08) > apelin-13(R10M) (pD2 = 8.26±0.17) > apelin-13(F13A) (pD2 = 7.98±0.04)  [Pyr1]apelin-13(1-12) (pD2 = 7.84±0.06) >> shorter fragments (pD2<6). [Pyr1]apelin-13(1-12) and apelin-13(F13A) contracted human saphenous vein with similar sub-nanomolar potencies and [Pyr1]apelin-13(1-12) was a potent inotrope in paced mouse right ventricle and human atria. [Pyr1]apelin-13(1-12) elicited a dose-dependent decrease in blood pressure in anaesthetized rat and dose-dependent increase in forearm blood flow in human volunteers. Conclusions: We provide evidence that ACE2 cleaves [Pyr1]apelin-13 to [Pyr1]apelin-13(1-12) and this cleavage product is expressed in human Cardiovascular tissues. We have demonstrated biological activity of [Pyr1]apelin-13(1-12) at the human and rodent apelin receptor in vitro and in vivo. Our data show that reported enhanced ACE2 activity in Cardiovascular disease should not significantly compromise the beneficial effects of apelin based therapies for example in PAH.