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Neuregulin-1 attenuates hemolysis- and ischemia induced-cerebrovascular inflammation associated with sickle cell disease

  • Christopher Chambliss
    Correspondence
    Corresponding authors.
    Affiliations
    Pediatrics Institute, Emory University School of Medicine, 2015 Uppergate Drive, Atlanta, GA 30322, United States

    Aflac Cancer and Blood Disorders Center, 2015 Uppergate Drive, Atlanta, GA 30322, United States

    Cardiovascular Research Institute, Morehouse School of Medicine, 720 Westview Drive SW, Atlanta, GA 30310, United States
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  • Jonathan K. Stiles
    Correspondence
    Corresponding authors.
    Affiliations
    Department of Microbiology, Biochemistry, and Immunology, Morehouse School of Medicine, 720 Westview Drive SW, Atlanta, GA 30310, United States
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  • Beatrice E. Gee
    Affiliations
    Pediatrics Institute, Emory University School of Medicine, 2015 Uppergate Drive, Atlanta, GA 30322, United States

    Aflac Cancer and Blood Disorders Center, 2015 Uppergate Drive, Atlanta, GA 30322, United States

    Children's Healthcare of Atlanta, 35 Jesse Hill Jr Drive SE, Atlanta, GA 30303, United States

    Department of Pediatrics, Morehouse School of Medicine, 720 Westview Drive SW, Atlanta, GA 30310, United States
    Search for articles by this author

      Highlights

      • Mice with SCD show elevated plasma NRG-1 levels in comparison to healthy controls.
      • Experimental conditions mimicking hemolysis and ischemia increase NRG-1 production.
      • NRG-1 reduced inflammatory cytokine and adhesion molecule expression.
      • NRG-1 expression in SCD is likely a protective endogenous response.

      Abstract

      Objectives

      Individuals with sickle cell disease (SCD) are at severely heightened risk for cerebrovascular injury and acute cerebrovascular events, including ischemic and hemorrhagic stroke, potentially leading to impaired development and life-long physical and cognitive disabilities. Cerebrovascular injury specific to SCD includes inflammation caused by underlying conditions of chronic hemolysis and reduced cerebrovascular perfusion. The objectives of this study were to investigate whether expression of neuregulin-1β (NRG-1), an endogenous neuroprotective polypeptide, is increased in SCD or experimental conditions mimicking the hemolysis and ischemic conditions of SCD, and to determine if treatment with exogenous NRG-1 reduces markers of cerebrovascular inflammation.

      Materials and methods

      Plasma and brain-specific NRG-1 levels were measured in transgenic SCD mice. Endogenous NRG-1 levels and response to experimental conditions of excess heme and ischemia were measured in cultured human brain microvascular cells and astrocytes. Pre-treatment with NRG-1 was used to determine NRG-1’s ability to ameliorate resultant cerebrovascular inflammation.

      Results

      Plasma and brain-specific NRG-1 were elevated in transgenic SCD mice compared to healthy controls. Neuregulin-1 expression was significantly increased in cultured human microvascular cells and astrocytes exposed to excess heme and ischemia. Pre-treatment with NRG-1 reduced inflammatory chemokine (CXCL-1 and CXCL-10) and adhesion molecule (ICAM-1 and VCAM-1) expression and increased pro-angiogenic factors (VEGF-A) in microvascular cells and astrocytes exposed to excess heme and ischemia.

      Conclusions

      Elevated NRG-1 in SCD is likely a protective endogenous response to ongoing cerebrovascular insults caused by chronic hemolysis and reduced cerebrovascular perfusion. Administration of NRG-1 to reduce cerebrovascular inflammation may be therapeutically beneficial in SCD and warrants continued investigation.

      Keywords

      Abbreviations:

      AA (Hemoglobin AA), AB (Antibody), AC (Hemoglobin AC), AKT (Protein kinase B), ARC (Absolute reticulocyte count), AS (Hemoglobin AS), BDNF (Brain derived neurotrophic factor), CT (Cycle of threshold), CXCL-1 (C-X-C motif chemokine ligand 1), CXCL-10 (C-X-C motif chemokine ligand 10), DAPI (4,6-diamidino-2-phenylindole), DMSO (Dimethyl sulfoxide), ELISA (Enzyme-linked immunosorbent assay), ERBB4 (Erythroblastic oncogene B receptor tyrosine kinase 4), eNOS (Endothelial derived nitric oxide synthase), GFAP (Glial fibrillary acidic protein), HB (Hemoglobin), HBMVEC (Human brain microvascular endothelial cells), HO-1 (Heme oxygenase 1), GAPDH (Glyceraldehyde 3-phosphate dehydrogenase), GLU (Glutamine), ICAM-1 (Intracellular adhesion molecule 1), IQR (Interquartile range), LYS (Lysine), MCAO (Middle cerebral artery occlusion), N/A (Not applicable), NEUN (Neuronal nuclei), NRG-1 (Neuregulin-1), PBS (Phosphate buffered saline), PDGF-AA (Platelet derived growth factor type-aa), PGK-1 (Phosphoglycerate kinase 1), RBC (Red blood cell), RPM (Rotations per minute), SCA (Sickle cell anemia), SCD (Sickle cell disease), SS (Hemoglobin SS), STAT (Signal transducer and activator of transcription proteins), TCD (Transcranial Doppler), VAL (Valine), VEGFA (Vascular endothelial growth factor a), VCAM-1 (Vascular cell adhesion molecule 1), WBC (White blood cell)
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