Haemoglobin causes neuronal damage in vivo which is preventable by haptoglobin

Abstract After subarachnoid haemorrhage, prolonged exposure to toxic extracellular haemoglobin occurs in the brain. Here, we investigate the role of haemoglobin neurotoxicity in vivo and its prevention. In humans after subarachnoid haemorrhage, haemoglobin in cerebrospinal fluid was associated with neurofilament light chain, a marker of neuronal damage. Most haemoglobin was not complexed with haptoglobin, an endogenous haemoglobin scavenger present at very low concentration in the brain. Exogenously added haptoglobin bound most uncomplexed haemoglobin, in the first 2 weeks after human subarachnoid haemorrhage, indicating a wide therapeutic window. In mice, the behavioural, vascular, cellular and molecular changes seen after human subarachnoid haemorrhage were recapitulated by modelling a single aspect of subarachnoid haemorrhage: prolonged intrathecal exposure to haemoglobin. Haemoglobin-induced behavioural deficits and astrocytic, microglial and synaptic changes were attenuated by haptoglobin. Haptoglobin treatment did not attenuate large-vessel vasospasm, yet improved clinical outcome by restricting diffusion of haemoglobin into the parenchyma and reducing small-vessel vasospasm. In summary, haemoglobin toxicity is of clinical importance and preventable by haptoglobin, independent of large-vessel vasospasm.

[1]  V. Talesa,et al.  Oleuropein-Induced Apoptosis Is Mediated by Mitochondrial Glyoxalase 2 in NSCLC A549 Cells: A Mechanistic Inside and a Possible Novel Nonenzymatic Role for an Ancient Enzyme , 2019, Oxidative medicine and cellular longevity.

[2]  D. Bulters,et al.  Neuroprotective Role of the Nrf2 Pathway in Subarachnoid Haemorrhage and Its Therapeutic Potential , 2019, Oxidative medicine and cellular longevity.

[3]  S. Blackburn,et al.  Haptoglobin genotype and aneurysmal subarachnoid hemorrhage , 2019, Neurology.

[4]  J. Jeon,et al.  Clazosentan for Aneurysmal Subarachnoid Hemorrhage: An Updated Meta-Analysis with Trial Sequential Analysis. , 2019, World neurosurgery.

[5]  S. Blackburn,et al.  Haemoglobin scavenging in intracranial bleeding: biology and clinical implications , 2018, Nature Reviews Neurology.

[6]  Yang Cao,et al.  Hemopexin increases the neurotoxicity of hemoglobin when haptoglobin is absent , 2018, Journal of neurochemistry.

[7]  R. Keep,et al.  Association of Brain CD163 Expression and Brain Injury/Hydrocephalus Development in a Rat Model of Subarachnoid Hemorrhage , 2018, Front. Neurosci..

[8]  D. Stein,et al.  Neurobehavioral testing in subarachnoid hemorrhage: A review of methods and current findings in rodents , 2017, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[9]  M. Meyerand,et al.  Intrathecal antibody distribution in the rat brain: surface diffusion, perivascular transport and osmotic enhancement of delivery , 2017, The Journal of physiology.

[10]  G. Zipfel,et al.  A novel fluorescent imaging technique for assessment of cerebral vasospasm after experimental subarachnoid hemorrhage , 2017, Scientific Reports.

[11]  Terrance T. Kummer,et al.  Minimal Long-Term Neurobehavioral Impairments after Endovascular Perforation Subarachnoid Hemorrhage in Mice , 2017, Scientific Reports.

[12]  J. ReederBrandon,et al.  Redox and Peroxidase Activities of the Hemoglobin Superfamily: Relevance to Health and Disease. , 2017, Antioxidants & redox signaling.

[13]  R. Keep,et al.  CD163 Expression in Neurons After Experimental Intracerebral Hemorrhage , 2017, Stroke.

[14]  S. Moestrup,et al.  The absence of the CD163 receptor has distinct temporal influences on intracerebral hemorrhage outcomes , 2017, Journal of Cerebral Blood Flow and Metabolism.

[15]  R. L. Macdonald,et al.  Spontaneous subarachnoid haemorrhage , 2017, The Lancet.

[16]  R. Regan,et al.  Haptoglobin increases the vulnerability of CD163‐expressing neurons to hemoglobin , 2016, Journal of neurochemistry.

[17]  R. Du,et al.  Association between S100B Levels and Long-Term Outcome after Aneurysmal Subarachnoid Hemorrhage: Systematic Review and Pooled Analysis , 2016, PloS one.

[18]  R. Keep,et al.  Hemoglobin-induced neuronal degeneration in the hippocampus after neonatal intraventricular hemorrhage , 2016, Brain Research.

[19]  D. Bulters,et al.  Heme–Hemopexin Scavenging Is Active in the Brain and Associates With Outcome After Subarachnoid Hemorrhage , 2016, Stroke.

[20]  N. Plesnila,et al.  Are We Barking Up the Wrong Vessels?: Cerebral Microcirculation After Subarachnoid Hemorrhage , 2015, Stroke.

[21]  F. Heppner,et al.  Microglia inflict delayed brain injury after subarachnoid hemorrhage , 2015, Acta Neuropathologica.

[22]  J. Vickers,et al.  Diffuse axonal injury in brain trauma: insights from alterations in neurofilaments , 2014, Front. Cell. Neurosci..

[23]  H. Lutz,et al.  Mechanisms tagging senescent red blood cells for clearance in healthy humans , 2013, Front. Physiol..

[24]  P. Buehler,et al.  Human Hp1-1 and Hp2-2 phenotype-specific haptoglobin therapeutics are both effective in vitro and in guinea pigs to attenuate hemoglobin toxicity. , 2013, Antioxidants & redox signaling.

[25]  V. Perry,et al.  Immune complex formation impairs the elimination of solutes from the brain: implications for immunotherapy in Alzheimer’s disease , 2013, Acta neuropathologica communications.

[26]  S. Moestrup,et al.  Plasma clearance of hemoglobin and haptoglobin in mice and effect of CD163 gene targeting disruption. , 2013, Antioxidants & redox signaling.

[27]  K. Blennow,et al.  Idiopathic normal-pressure hydrocephalus , 2013, Neurology.

[28]  A. Zlotnik,et al.  The neuro-behavioral profile in rats after subarachnoid hemorrhage , 2013, Brain Research.

[29]  J. Bertolini,et al.  Production of Plasma Proteins for Therapeutic Use , 2012 .

[30]  V. Perry,et al.  Soluble Axoplasm Enriched from Injured CNS Axons Reveals the Early Modulation of the Actin Cytoskeleton , 2012, PloS one.

[31]  I. Galea,et al.  The intrathecal CD163-haptoglobin–hemoglobin scavenging system in subarachnoid hemorrhage , 2012, Journal of neurochemistry.

[32]  B. Reeder.  The redox activity of hemoglobins: from physiologic functions to pathologic mechanisms. , 2010, Antioxidants & redox signaling.

[33]  J. Fandino,et al.  Standard intracranial in vivo animal models of delayed cerebral vasospasm , 2010, British journal of neurosurgery.

[34]  T. Schweizer,et al.  Cognitive and Functional Outcome After Aneurysmal Subarachnoid Hemorrhage , 2010, Stroke.

[35]  I. Cuthill,et al.  Reporting : The ARRIVE Guidelines for Reporting Animal Research , 2010 .

[36]  V. Verhasselt,et al.  Breast milk immune complexes are potent inducers of oral tolerance in neonates and prevent asthma development , 2010, Mucosal Immunology.

[37]  Lello Zolla,et al.  Blood-related proteomics. , 2010, Journal of proteomics.

[38]  J. Grotta,et al.  Neuroprotective Role of Haptoglobin after Intracerebral Hemorrhage , 2009, The Journal of Neuroscience.

[39]  R. Macdonald,et al.  Anterior circulation mouse model of subarachnoid hemorrhage , 2009, Brain Research.

[40]  J. Stark,et al.  Coronary intraplaque hemorrhage evokes a novel atheroprotective macrophage phenotype. , 2009, The American journal of pathology.

[41]  V. Perry,et al.  Immune-to-brain signalling: The role of cerebral CD163-positive macrophages , 2008, Neuroscience Letters.

[42]  P. Buehler,et al.  The reaction of hydrogen peroxide with hemoglobin induces extensive alpha-globin crosslinking and impairs the interaction of hemoglobin with endogenous scavenger pathways. , 2008, Free radical biology & medicine.

[43]  A. Levy,et al.  Rate of nitric oxide scavenging by hemoglobin bound to haptoglobin. , 2008, Nitric oxide : biology and chemistry.

[44]  T. Beems,et al.  Increased GFAP and S100β but not NSE serum levels after subarachnoid haemorrhage are associated with clinical severity , 2006, European journal of neurology.

[45]  P. Højrup,et al.  Identification of the receptor scavenging hemopexin-heme complexes. , 2005, Blood.

[46]  A. Levy,et al.  Haptoglobin Genotype– and Diabetes-Dependent Differences in Iron-Mediated Oxidative Stress In Vitro and In Vivo , 2005, Circulation research.

[47]  S. Moestrup,et al.  Molecular Characterization of the Haptoglobin·Hemoglobin Receptor CD163 , 2004, Journal of Biological Chemistry.

[48]  Y. Imai,et al.  Microglia/macrophage‐specific protein Iba1 binds to fimbrin and enhances its actin‐bundling activity , 2004, Journal of neurochemistry.

[49]  V. Perry,et al.  Synaptic changes characterize early behavioural signs in the ME7 model of murine prion disease , 2003, The European journal of neuroscience.

[50]  D. Hanley,et al.  Blood Clot Resolution in Human Cerebrospinal Fluid: Evidence of First-order Kinetics , 2001, Neurosurgery.

[51]  M. Wilson,et al.  Toxicity of myoglobin and haemoglobin: oxidative stress in patients with rhabdomyolysis and subarachnoid haemorrhage. , 2001, Biochemical Society transactions.

[52]  Robert W. Williams,et al.  Complex trait analysis of the mouse striatum: independent QTLs modulate volume and neuron number , 2001, BMC Neuroscience.

[53]  E. Lentjes,et al.  Iterative model for the calculation of oxyhemoglobin, methemoglobin, and bilirubin in absorbance spectra of cerebrospinal fluid. , 2001, Clinical chemistry.

[54]  R. Macdonald,et al.  Hemoglobin increases endothelin-1 in endothelial cells by decreasing nitric oxide. , 2001, Biochemical and biophysical research communications.

[55]  S. Moestrup,et al.  Identification of the haemoglobin scavenger receptor , 2001, Nature.

[56]  Hugh Davson,et al.  Physiology of the CSF and Blood-Brain Barriers , 1996 .

[57]  J. Costentin,et al.  Thigmotaxis as an index of anxiety in mice. Influence of dopaminergic transmissions , 1994, Behavioural Brain Research.

[58]  R O Weller,et al.  CSF drains directly from the subarachnoid space into nasal lymphatics in the rat. Anatomy, histology and immunological significance , 1993, Neuropathology and applied neurobiology.

[59]  S. Panter,et al.  Neurotoxicity of hemoglobin in cortical cell culture , 1993, Neuroscience Letters.

[60]  C. Morris,et al.  Evidence for the localization of haemopexin immunoreactivity in neurones in the human brain , 1993, Neuroscience Letters.

[61]  S. Moestrup,et al.  Distribution of the α2-macroglobulin receptor/low density lipoprotein receptor-related protein in human tissues , 1992, Cell and Tissue Research.

[62]  V. Perry,et al.  The acute inflammatory response to lipopolysaccharide in cns parenchyma differs from that in other body tissues , 1992, Neuroscience.

[63]  A. Koeppen,et al.  Experimental Superficial Siderosis of the Central Nervous System. I. Morphological Observations , 1991, Journal of neuropathology and experimental neurology.

[64]  G. Murray,et al.  Effect of oral nimodipine on cerebral infarction and outcome after subarachnoid haemorrhage: British aneurysm nimodipine trial. , 1989, BMJ.

[65]  G. Teasdale,et al.  A universal subarachnoid hemorrhage scale: report of a committee of the World Federation of Neurosurgical Societies. , 1988, Journal of neurology, neurosurgery, and psychiatry.

[66]  T. Kojima,et al.  Endothelium-dependent relaxation of canine basilar arteries. Part 2: Inhibition by hemoglobin and cerebrospinal fluid from patients with aneurysmal subarachnoid hemorrhage. , 1987, Stroke.

[67]  R. Rudick,et al.  Clearance of albumin from mouse subarachnoid space: a measure of CSF bulk flow , 1982, Journal of Neuroscience Methods.

[68]  P. Gray,et al.  Stress-induced responses and open-field behavior in estrous and nonestrous mice , 1982, Physiology & Behavior.

[69]  C. Patlak,et al.  Efflux of radiolabeled polyethylene glycols and albumin from rat brain. , 1981, The American journal of physiology.

[70]  A. D. Rosen,et al.  Focal epileptogenesis after intracortical hemoglobin injection , 1979, Experimental Neurology.

[71]  A. Gressner,et al.  Haptoglobin , 2019, Springer Reference Medizin.

[72]  Michael Detmar,et al.  A dural lymphatic vascular system that drains brain interstitial fluid and macromolecules , 2015 .

[73]  ANTIOXlDANTSREDOXS1GNALlNG,et al.  Antioxidants ＆ redox signaling , 2013 .

[74]  R. Macdonald,et al.  Vasospasm: Measurement of Diameter, Perimeter, and Wall Thickness , 2012 .

[75]  S. Moestrup,et al.  Tumor necrosis factor -converting enzyme (TACE/ADAM17) mediates ectodomain shedding of the scavenger receptor CD163 , 2010 .

[76]  Jürgen Götz,et al.  Primary support cultures of hippocampal and substantia nigra neurons , 2008, Nature Protocols.

[77]  V. Perry,et al.  What is immune privilege (not)? , 2007, Trends in immunology.

[78]  M Cortina Borja,et al.  The Design of Animal Experiments: Reducing the use of animals in research through better experimental design. (Revised and updated edition) , 2002 .

[79]  Y. Benjamini,et al.  Controlling the false discovery rate: a practical and powerful approach to multiple testing , 1995 .