Molecular Medicine

, Volume 20, Issue 1, pp 435–447 | Cite as

Chemokine (C-X-C Motif) Receptor 4 and Atypical Chemokine Receptor 3 Regulate Vascular α1-Adrenergic Receptor Function

  • Harold H. BachIV
  • Yee M. Wong
  • Abhishek Tripathi
  • Amanda M. Nevins
  • Richard L. Gamelli
  • Brian F. Volkman
  • Kenneth L. Byron
  • Matthias Majetschak
Research Article


Chemokine (C-X-C motif) receptor (CXCR) 4 and atypical chemokine receptor (ACKR) 3 ligands have been reported to modulate cardiovascular function in various disease models. The underlying mechanisms, however, remain unknown. Thus, it was the aim of the present study to determine how pharmacological modulation of CXCR4 and ACKR3 regulate cardiovascular function. In vivo administration of TC14012, a CXCR4 antagonist and ACKR3 agonist, caused cardiovascular collapse in normal animals. During the cardiovascular stress response to hemorrhagic shock, ubiquitin, a CXCR4 agonist, stabilized blood pressure, whereas coactivation of CXCR4 and ACKR3 with CXC chemokine ligand 12 (CXCL12), or blockade of CXCR4 with AMD3100 showed opposite effects. While CXCR4 and ACKR3 ligands did not affect myocardial function, they selectively altered vascular reactivity upon α1-adrenergic receptor (AR) activation in pressure myography experiments. CXCR4 activation with ubiquitin enhanced α1-AR-mediated vasoconstriction, whereas ACKR3 activation with various natural and synthetic ligands antagonized α1-AR-mediated vasoconstriction. The opposing effects of CXCR4 and ACKR3 activation by CXCL12 could be dissected pharmacologically. CXCR4 and ACKR3 ligands did not affect vasoconstriction upon activation of voltage-operated Ca2+ channels or endothelin receptors. Effects of CXCR4 and ACKR3 agonists on vascular α1-AR responsiveness were independent of the endothelium. These findings suggest that CXCR4 and ACKR3 modulate α1-AR reactivity in vascular smooth muscle and regulate hemodynamics in normal and pathological conditions. Our observations point toward CXCR4 and ACKR3 as new pharmacological targets to control vasore-activity and blood pressure.



The authors thank Heather M La Porte for technical help and P de Tombe, X Ji, S Sadayappan, and R. Tiniakov, Loyola University Chicago, for help with myocardial function analyses. This research was made possible, in part, by a grant that was awarded and administered by the U.S. Army Medical Research & Materiel Command (USAMRMC) and the Telemedicine and Advanced Technology Research Center (TATRC), at Fort Detrick, MD, USA, under contract number W81XWH1020122. The views, opinions and/or findings contained in this research are those of the author(s) and do not necessarily reflect the views of the Department of Defense and should not be construed as an official DoD/Army position, policy or decision unless so designated by other documentation. No official endorsement should be made. This research was also supported, in part, by grants from the American Heart Association (13GRNT17230072), the NIH (T32GM008750) and the Dr. Ralph and Marian Falk Medical Research Trust.

Supplementary material

10020_2014_2001435_MOESM1_ESM.pdf (1.4 mb)
Supplementary material, approximately 1454 KB.


  1. 1.
    Bachelerie F, et al. (2013) International Union of Basic and Clinical Pharmacology. [corrected]. LXXXIX. Update on the extended family of chemokine receptors and introducing a new nomenclature for atypical chemokine receptors. Pharmacol. Rev. 66:1–79; erratum 66:467.CrossRefPubMedGoogle Scholar
  2. 2.
    Tachibana K, et al. (1998) The chemokine receptor CXCR4 is essential for vascularization of the gastrointestinal tract. Nature. 393:591–4.CrossRefPubMedGoogle Scholar
  3. 3.
    Gerrits H, et al. (2008) Early postnatal lethality and cardiovascular defects in CXCR7-deficient mice. Genesis. 46:235–45.CrossRefPubMedGoogle Scholar
  4. 4.
    Nagasawa T, et al. (1996) Defects of B-cell lymphopoiesis and bone-marrow myelopoiesis in mice lacking the CXC chemokine PBSF/SDF-1. Nature. 382:635–8.CrossRefPubMedGoogle Scholar
  5. 5.
    Sierro F, et al. (2007) Disrupted cardiac development but normal hematopoiesis in mice deficient in the second CXCL12/SDF-1 receptor, CXCR7. Proc. Natl. Acad. Sci. U. S. A. 104:14759–64.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Bleul CC, et al. (1996) The lymphocyte chemoattractant SDF-1 is a ligand for LESTR/fusin and blocks HIV-1 entry. Nature. 382:829–33.CrossRefPubMedGoogle Scholar
  7. 7.
    Balabanian K, et al. (2005) The chemokine SDF-1/CXCL12 binds to and signals through the orphan receptor RDC1 in T lymphocytes. J. Biol. Chem. 280:35760–6.CrossRefPubMedGoogle Scholar
  8. 8.
    Teicher BA, Fricker SP. (2010) CXCL12 (SDF-1)/CXCR4 pathway in cancer. Clin. Cancer Res. 16:2927–31.CrossRefPubMedGoogle Scholar
  9. 9.
    Nagasawa T, Tachibana K, Kishimoto T. (1998) A novel CXC chemokine PBSF/SDF-1 and its receptor CXCR4: their functions in development, hematopoiesis and HIV infection. Semin. Immunol. 10:179–85.CrossRefPubMedGoogle Scholar
  10. 10.
    Rajagopal S, et al. (2010) Beta-arrestin- but not G protein-mediated signaling by the “decoy” receptor CXCR7. Proc. Natl. Acad. Sci. U. S. A. 107:628–32.CrossRefPubMedGoogle Scholar
  11. 11.
    Boldajipour B, et al. (2008) Control of chemokineguided cell migration by ligand sequestration. Cell. 132:463–73.CrossRefPubMedGoogle Scholar
  12. 12.
    Kumar R, et al. (2012) CXCR7 mediated Gia independent activation of ERK and Akt promotes cell survival and chemotaxis in T cells. Cell. Immunol. 272:230–41.CrossRefPubMedGoogle Scholar
  13. 13.
    Regard JB, Sato IT, Coughlin SR. (2008) Anatomical profiling of G protein-coupled receptor expression. Cell. 135:561–71.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Tripathi A, Davis JD, Staren DM, Volkman BF, Majetschak M. (2013) CXC chemokine receptor 4 signaling upon co-activation with stromal cell-derived factor-1alpha and ubiquitin. Cytokine. 65:121–5.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    LaRocca TJ, et al. (2010) β2-Adrenergic receptor signaling in the cardiac myocyte is modulated by interactions with CXCR4. J. Cardiovasc. Pharmacol. 56:548–59.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Agarwal U, et al. (2010) Role of cardiac myocyte CXCR4 expression in development and left ventricular remodeling after acute myocardial infarction. Circ. Res. 107:667–76.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Levoye A, Balabanian K, Baleux F, Bachelerie F, Lagane B. (2009) CXCR7 heterodimerizes with CXCR4 and regulates CXCL12-mediated G protein signaling. Blood. 113:6085–93.CrossRefPubMedGoogle Scholar
  18. 18.
    Saini V, Marchese A, Majetschak M. (2010) CXC chemokine receptor 4 is a cell surface receptor for extracellular ubiquitin. J. Biol. Chem. 285:15566–76.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Majetschak M, Cohn SM, Nelson JA, Burton EH, Obertacke U, Proctor KG. (2004) Effects of exogenous ubiquitin in lethal endotoxemia. Surgery. 135:536–43.CrossRefPubMedGoogle Scholar
  20. 20.
    Baker TA, Romero J, Bach HHt, Strom JA, Gamelli RL, Majetschak M. (2012) Effects of exogenous ubiquitin in a polytrauma model with blunt chest trauma. Crit. Care Med. 40:2376–84.CrossRefPubMedGoogle Scholar
  21. 21.
    Bach Iv HH, Saini V, Baker TA, Tripathi A, Gamelli RL, Majetschak M. (2012) Initial assessment of the role of CXC chemokine receptor 4 after polytrauma. Mol. Med. 18:1056–66.Google Scholar
  22. 22.
    Bodart V, et al. (2009) Pharmacology of AMD3465: a small molecule antagonist of the chemokine receptor CXCR4. Biochem. Pharmacol. 78:993–1000.CrossRefPubMedGoogle Scholar
  23. 23.
    Chu PY, et al. (2011) CXCR4 antagonism attenuates the cardiorenal consequences of mineralocorticoid excess. Circ. Heart Fail. 4:651–8.CrossRefPubMedGoogle Scholar
  24. 24.
    Yu L, Hales CA. (2011)Effect of chemokine receptor CXCR4 on hypoxia-induced pulmonary hypertension and vascular remodeling in rats. Respir. Res. 12:21.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Zabel BA, et al. (2009) Elucidation of CXCR7-mediated signaling events and inhibition of CXCR4-mediated tumor cell transendothelial migration by CXCR7 ligands. J. Immunol. 183:3204–11.CrossRefPubMedGoogle Scholar
  26. 26.
    Sartina E, et al. (2012) Antagonism of CXCR7 attenuates chronic hypoxia-induced pulmonary hypertension. Pediatr. Res. 71:682–8.CrossRefPubMedGoogle Scholar
  27. 27.
    Veldkamp CT, Seibert C, Peterson FC, Sakmar TP, Volkman BF. (2006) Recognition of a CXCR4 sulfotyrosine by the chemokine stromal cell-derived factor-1alpha (SDF-1alpha/CXCL12). J. Mol. Biol. 359:1400–9.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Veldkamp CT, et al. (2008) Structural basis of CXCR4 sulfotyrosine recognition by the chemokine SDF-1/CXCL12. Sci. Signal. 1:ra4.CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Veldkamp CT, et al. (2009) Monomeric structure of the cardioprotective chemokine SDF-1/CXCL12. Protein Sci. 18:1359–69.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Takekoshi T, Ziarek JJ, Volkman BF, Hwang ST. (2012) A locked, dimeric CXCL12 variant effectively inhibits pulmonary metastasis of CXCR4-expressing melanoma cells due to enhanced serum stability. Mol. Cancer Ther. 11:2516–25.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    (1985) Bacterial endotoxins/pyrogens [Internet]. Silver Spring (MD): FDA; [cited 2014 Aug 29]. (Inspection technical guide). Available from: (web page last updated 2009 Feb 17).
  32. 32.
    Committee for the Update of the Guide for the Care and Use of Laboratory Animals, Institute for Laboratory Animal Research, Division on Earth and Life Studies, National Research Council of the National Academies. (2011) Guide for the Care and Use of Laboratory Animals. 8th edition. Washington (DC): National Academies Press.Google Scholar
  33. 33.
    Henderson KK, Byron KL. (2007) Vasopressin-induced vasoconstriction: two concentration-dependent signaling pathways. J. Appl. Physiol. 102:1402–9.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Brueggemann LI, Mackie AR, Mani BK, Cribbs LL, Byron KL. (2009) Differential effects of selective cyclooxygenase-2 inhibitors on vascular smooth muscle ion channels may account for differences in cardiovascular risk profiles. Mol. Pharmacol. 76:1053–61.CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Brueggemann LI, Mani BK, Haick J, Byron KL. (2012) Exploring arterial smooth muscle Kv7 potassium channel function using patch clamp electrophysiology and pressure myography. J. Vis. Exp. 14:e4263.Google Scholar
  36. 36.
    Mani BK, Brueggemann LI, Cribbs LL, Byron KL. (2011) Activation of vascular KCNQ (Kv7) potassium channels reverses spasmogen-induced constrictor responses in rat basilar artery. Br. J. Pharmacol. 164:237–49.CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Dow JW, Harding NG, Powell T. (1981) Isolated cardiac myocytes. I. Preparation of adult myocytes and their homology with the intact tissue. Cardiovasc. Res. 15:483–514.CrossRefPubMedGoogle Scholar
  38. 38.
    Govindan S, et al. (2012) Pathogenic properties of the N-terminal region of cardiac myosin binding protein-C in vitro. J. Muscle Res. Cell Motil. 33:17–30.CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Jin CZ, et al. (2013) Myofilament Ca desensitization mediates positive lusitropic effect of neuronal nitric oxide synthase in left ventricular myocytes from murine hypertensive heart. J. Mol. Cell. Cardiol. 60:107–15.CrossRefPubMedGoogle Scholar
  40. 40.
    Geng Q, et al. (2009) A subset of 26S proteasomes is activated at critically low ATP concentrations and contributes to myocardial injury during cold ischemia. Biochem. Biophys. Res. Commun. 390:1136–41.CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Bach HHt, Laporte HM, Wong YM, Gamelli RL, Majetschak M. (2013) Proteasome inhibition prolongs survival during lethal hemorrhagic shock in rats. J. Trauma Acute Care Surg. 74:499–507.CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Saini V, et al. (2011) The CXC chemokine receptor 4 ligands ubiquitin and stromal cell-derived factor-1α function through distinct receptor interactions. J. Biol. Chem. 286:33466–77.CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Hatse S, Princen K, Bridger G, De Clercq E, Schols D. (2002) Chemokine receptor inhibition by AMD3100 is strictly confined to CXCR4. FEBS Lett. 527:255–62.CrossRefPubMedGoogle Scholar
  44. 44.
    Gravel S, et al. (2010) The peptidomimetic CXCR4 antagonist TC14012 recruits beta-arrestin to CXCR7: roles of receptor domains. J. Biol. Chem. 285:37939–43.CrossRefPubMedPubMedCentralGoogle Scholar
  45. 45.
    Majetschak M, Cohn SM, Obertacke U, Proctor KG. (2004) Therapeutic potential of exogenous ubiquitin during resuscitation from severe trauma. J. Trauma. 56:991–9.CrossRefPubMedGoogle Scholar
  46. 46.
    O’Boyle G, Mellor P, Kirby JA, Ali S. (2009) Anti-inflammatory therapy by intravenous delivery of non-heparan sulfate-binding CXCL12. FASEB J. 23:3906–16.CrossRefPubMedPubMedCentralGoogle Scholar
  47. 47.
    Misra P, et al. (2008) Quantitation of CXCR4 expression in myocardial infarction using 99mTc-labeled SDF-1alpha. J. Nucl. Med. 49:963–9.CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Burns JM, et al. (2006) A novel chemokine receptor for SDF-1 and I-TAC involved in cell survival, cell adhesion, and tumor development. J. Exp. Med. 203:2201–13.CrossRefPubMedPubMedCentralGoogle Scholar
  49. 49.
    Berahovich RD, Penfold ME, Schall TJ. (2010) Nonspecific CXCR7 antibodies. Immunol. Lett. 133:112–4.CrossRefPubMedGoogle Scholar
  50. 50.
    Majetschak M. (2011) Extracellular ubiquitin: immune modulator and endogenous opponent of damage-associated molecular pattern molecules. J. Leukoc. Biol. 89:205–19.CrossRefPubMedGoogle Scholar
  51. 51.
    Butera D, et al. (2005) Plasma chemokine levels correlate with the outcome of antiviral therapy in patients with hepatitis C. Blood. 106:1175–82.CrossRefPubMedPubMedCentralGoogle Scholar
  52. 52.
    Derdeyn CA, et al. (1999) Correlation between circulating stromal cell-derived factor 1 levels and CD4+ cell count in human immunodeficiency virus type 1-infected individuals. AIDS Res. Hum. Retroviruses. 15:1063–71.CrossRefPubMedGoogle Scholar
  53. 53.
    Kieffer AE, et al. (2003) The N- and C-terminal fragments of ubiquitin are important for the antimicrobial activities. Faseb J. 17:776–8.CrossRefPubMedGoogle Scholar
  54. 54.
    Hendrix CW, et al. (2000) Pharmacokinetics and safety of AMD-3100, a novel antagonist of the CXCR-4 chemokine receptor, in human volunteers. Antimicrob. Agents Chemother. 44:1667–73.CrossRefPubMedPubMedCentralGoogle Scholar
  55. 55.
    Widney DP, Xia YR, Lusis AJ, Smith JB. (2000) The murine chemokine CXCL11 (IFN-inducible T cell alpha chemoattractant) is an IFN-gamma- and lipopolysaccharide-inducible glucocortioid-attenuated response gene expressed in lung and other tissues during endotoxemia. J. Immunol. 164:6322–31.CrossRefPubMedGoogle Scholar
  56. 56.
    Moore CA, Milano SK, Benovic JL. (2007) Regulation of receptor trafficking by GRKs and arrestins. Annu. Rev. Physiol. 69:451–82.CrossRefPubMedGoogle Scholar
  57. 57.
    Busillo JM, Benovic JL. (2007) Regulation of CXCR4 signaling. Biochim. Biophys. Acta. 1768:952–63.CrossRefPubMedGoogle Scholar
  58. 58.
    Marchese A, Chen C, Kim YM, Benovic JL. (2003) The ins and outs of G protein-coupled receptor trafficking. Trends Biochem. Sci. 28:369–76.CrossRefPubMedGoogle Scholar
  59. 59.
    Rockman HA, Koch WJ, Lefkowitz RJ. (2002) Seven-transmembrane-spanning receptors and heart function. Nature. 415:206–12.CrossRefGoogle Scholar
  60. 60.
    Diviani D, et al. (1996) Effect of different G protein-coupled receptor kinases on phosphorylation and desensitization of the alpha1B-adrenergic receptor. J. Biol. Chem. 271:5049–58.CrossRefPubMedGoogle Scholar
  61. 61.
    Collins S, Bouvier M, Lohse MJ, Benovic JL, Caron MG, Lefkowitz RJ. (1990) Mechanisms involved in adrenergic receptor desensitization. Biochem. Soc. Trans. 18:541–4.CrossRefPubMedGoogle Scholar
  62. 62.
    McGraw DW, et al. (2006) Airway smooth muscle prostaglandin-EP1 receptors directly modulate beta2-adrenergic receptors within a unique heterodimeric complex. J. Clin. Invest. 116:1400–9.CrossRefPubMedPubMedCentralGoogle Scholar
  63. 63.
    Dzimiri N. (2002) Receptor crosstalk. Implications for cardiovascular function, disease and therapy. Eur. J. Biochem. 269:4713–30.CrossRefGoogle Scholar
  64. 64.
    Milligan G, Canals M, Pediani JD, Ellis J, Lopez-Gimenez JF. (2006) The role of GPCR dimerisation/oligomerisation in receptor signalling. Ernst Schering Found. Symp. Proc. 2:145–61.Google Scholar
  65. 65.
    Stanasila L, Perez JB, Vogel H, Cotecchia S. (2003) Oligomerization of the alpha 1a- and alpha 1b-adrenergic receptor subtypes. Potential implications in receptor internalization. J. Biol. Chem. 278:40239–51.CrossRefPubMedGoogle Scholar
  66. 66.
    Mustafa S, et al. (2012) Identification and profiling of novel alpha1A-adrenoceptor-CXC chemokine receptor 2 heteromer. J. Biol. Chem. 287:12952–65.CrossRefPubMedPubMedCentralGoogle Scholar
  67. 67.
    Schols D, Struyf S, Van Damme J, Este JA, Henson G, De Clercq E. (1997) Inhibition of T-tropic HIV strains by selective antagonization of the chemokine receptor CXCR4. J. Exp. Med. 186:1383–8.CrossRefPubMedPubMedCentralGoogle Scholar
  68. 68.
    Kalatskaya I, Berchiche YA, Gravel S, Limberg BJ, Rosenbaum JS, Heveker N. (2009) AMD3100 is a CXCR7 ligand with allosteric agonist properties. Mol. Pharmacol. 75:1240–7.CrossRefPubMedGoogle Scholar

Copyright information

© The Author(s) 2014

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, and provide a link to the Creative Commons license. You do not have permission under this license to share adapted material derived from this article or parts of it.

The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

To view a copy of this license, visit (

Authors and Affiliations

  • Harold H. BachIV
    • 1
    • 2
  • Yee M. Wong
    • 1
  • Abhishek Tripathi
    • 1
  • Amanda M. Nevins
    • 3
  • Richard L. Gamelli
    • 1
  • Brian F. Volkman
    • 3
  • Kenneth L. Byron
    • 2
  • Matthias Majetschak
    • 1
    • 2
    • 4
  1. 1.Department of SurgeryLoyola University ChicagoMaywoodUSA
  2. 2.Department of Molecular Pharmacology and TherapeuticsLoyola University ChicagoMaywoodUSA
  3. 3.Department of BiochemistryMedical College of WisconsinMilwaukeeUSA
  4. 4.Loyola University ChicagoMaywoodUSA

Personalised recommendations