title: HMGB1 and thrombin mediate the blood-brain barrier dysfunction acting as biomarkers of neuroinflammation and progression to neurodegeneration in Alzheimer's disease.
abstract: The blood-brain barrier (BBB) dysfunction represents an early feature of Alzheimer's disease (AD) that precedes the hallmarks of amyloid beta (amyloid β) plaque deposition and neuronal neurofibrillary tangle (NFT) formation. A damaged BBB correlates directly with neuroinflammation involving microglial activation and reactive astrogliosis, which is associated with increased expression and/or release of high-mobility group box protein 1 (HMGB1) and thrombin. However, the link between the presence of these molecules, BBB damage, and progression to neurodegeneration in AD is still elusive. Therefore, we aimed to profile and validate non-invasive clinical biomarkers of BBB dysfunction and neuroinflammation to assess the progression to neurodegeneration in mild cognitive impairment (MCI) and AD patients.
  
  
  
  We determined the serum levels of various proinflammatory damage-associated molecules in aged control subjects and patients with MCI or AD using validated ELISA kits. We then assessed the specific and direct effects of such molecules on BBB integrity in vitro using human primary brain microvascular endothelial cells or a cell line.
  
  
  
  We observed a significant increase in serum HMGB1 and soluble receptor for advanced glycation end products (sRAGE) that correlated well with amyloid beta levels in AD patients (vs. control subjects). Interestingly, serum HMGB1 levels were significantly elevated in MCI patients compared to controls or AD patients. In addition, as a marker of BBB damage, soluble thrombomodulin (sTM) antigen, and activity were significantly (and distinctly) increased in MCI and AD patients. Direct in vitro BBB integrity assessment further revealed a significant and concentration-dependent increase in paracellular permeability to dextrans by HMGB1 or α-thrombin, possibly through disruption of zona occludins-1 bands. Pre-treatment with anti-HMGB1 monoclonal antibody blocked HMGB1 effects and leaving BBB integrity intact.
  
  
  
  Our current studies indicate that thrombin and HMGB1 are causal proximate proinflammatory mediators of BBB dysfunction, while sTM levels may indicate BBB endothelial damage; HMGB1 and sRAGE might serve as clinical biomarkers for progression and/or therapeutic efficacy along the AD spectrum.
authors.affiliation: pHLOGISTIX LLC, 4220 Shawnee Mission Parkway, Fairway, KS, 66205, USA.|Department of Neurology, University of Kansas Medical Center, 3901 Rainbow Blvd, Kansas City, KS, 66160, USA.
authors.forename: Barry W
authors.initials: BW
authors.lastname: Festoff
authors.affiliation: Department of Pharmaceutical Sciences, Texas Tech University Health Sciences Center, 1300 S. Coulter Street, Amarillo, TX, 79106, USA.
authors.forename: Ravi K
authors.initials: RK
authors.lastname: Sajja
authors.affiliation: Clinical Research Department, R&D, Diagnostica Stago, Gennevilliers, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: van Dreden
authors.affiliation: Department of Pharmaceutical Sciences, Texas Tech University Health Sciences Center, 1300 S. Coulter Street, Amarillo, TX, 79106, USA. luca.cucullo@ttuhsc.edu.
authors.forename: Luca
authors.identifier: 0000-0002-2827-7162
authors.initials: L
authors.lastname: Cucullo
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000544:Alzheimer Disease / Q000150:complications; D016229:Amyloid beta-Peptides / Q000097:blood; D000906:Antibodies / Q000494:pharmacology; D001812:Blood-Brain Barrier / Q000187:drug effects / Q000378:metabolism / Q000503:physiopathology*; D002199:Capillary Permeability / Q000502:physiology; D002478:Cells, Cultured; D060825:Cognitive Dysfunction / Q000150:complications; D018450:Disease Progression; D004660:Encephalitis / Q000209:etiology / Q000473:pathology*; D042783:Endothelial Cells / Q000378:metabolism; D005260:Female; D017127:Glycation End Products, Advanced / Q000097:blood; D024243:HMGB1 Protein / Q000276:immunology / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D008297:Male; D008875:Middle Aged; D009410:Nerve Degeneration / Q000209:etiology / Q000473:pathology*; D010446:Peptide Fragments / Q000097:blood; D013917:Thrombin / Q000378:metabolism*
chemical_list: D016229:Amyloid beta-Peptides; D000906:Antibodies; D017127:Glycation End Products, Advanced; D024243:HMGB1 Protein; D010446:Peptide Fragments; C075222:amyloid beta-protein (1-42); D013917:Thrombin
keywords: Biomarkers; Blood-brain barrier; Clinical; DAMPS; HMGB1; Neurodegeneration; Neuroinflammation; Permeability; Thrombin