title: The NOTCH3 score: a pre-clinical CADASIL biomarker in a novel human genomic NOTCH3 transgenic mouse model with early progressive vascular NOTCH3 accumulation.
abstract: BACKGROUND
  CADASIL (Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy) is a hereditary small vessel disease caused by mutations in the NOTCH3 gene, leading to toxic NOTCH3 protein accumulation in the small- to medium sized arterioles. The accumulation is systemic but most pronounced in the brain vasculature where it leads to clinical symptoms of recurrent stroke and dementia. There is no therapy for CADASIL, and therapeutic development is hampered by a lack of feasible clinical outcome measures and biomarkers, both in mouse models and in CADASIL patients. To facilitate pre-clinical therapeutic interventions for CADASIL, we aimed to develop a novel, translational CADASIL mouse model.
  
  
  RESULTS
  We generated transgenic mice in which we overexpressed the full length human NOTCH3 gene from a genomic construct with the archetypal c.544C > T, p.Arg182Cys mutation. The four mutant strains we generated have respective human NOTCH3 RNA expression levels of 100, 150, 200 and 350 % relative to endogenous mouse Notch3 RNA expression. Immunohistochemistry on brain sections shows characteristic vascular human NOTCH3 accumulation in all four mutant strains, with human NOTCH3 RNA expression levels correlating with age at onset and progression of NOTCH3 accumulation. This finding was the basis for developing the 'NOTCH3 score', a quantitative measure for the NOTCH3 accumulation load. This score proved to be a robust and sensitive method to assess the progression of NOTCH3 accumulation, and a feasible biomarker for pre-clinical therapeutic testing.
  
  
  CONCLUSIONS
  This novel, translational CADASIL mouse model is a suitable model for pre-clinical testing of therapeutic strategies aimed at delaying or reversing NOTCH3 accumulation, using the NOTCH3 score as a biomarker.
authors.affiliation: Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands. j.w.rutten@lumc.nl.
authors.forename: Julie W
authors.initials: JW
authors.lastname: Rutten
authors.affiliation: Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands. r.r.klever@lumc.nl.
authors.forename: Roselin R
authors.initials: RR
authors.lastname: Klever
authors.affiliation: Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands. i.m.hegeman@lumc.nl.
authors.forename: Ingrid M
authors.initials: IM
authors.lastname: Hegeman
authors.affiliation: Department of Radiology, Leiden University Medical Center, Leiden, The Netherlands. d.poole@lumc.nl.
authors.forename: Dana S
authors.initials: DS
authors.lastname: Poole
authors.affiliation: Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands. j.g.dauwerse@lumc.nl.|Department of Clinical Genetics, K5-R, Leiden University Medical Center, PO Box 9600, 2300, RC, Leiden, The Netherlands. j.g.dauwerse@lumc.nl.
authors.forename: Hans G
authors.initials: HG
authors.lastname: Dauwerse
authors.affiliation: Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands. l.a.m.broos@lumc.nl.
authors.forename: Ludo A M
authors.initials: LA
authors.lastname: Broos
authors.affiliation: Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands. c.breukel@lumc.nl.
authors.forename: Cor
authors.initials: C
authors.lastname: Breukel
authors.affiliation: Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands. a.m.aartsma-rus@lumc.nl.
authors.forename: Annemieke M
authors.initials: AM
authors.lastname: Aartsma-Rus
authors.affiliation: Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands. j.s.verbeek@lumc.nl.
authors.forename: J Sjef
authors.initials: JS
authors.lastname: Verbeek
authors.affiliation: Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands. L.van_der_Weerd@lumc.nl.|Department of Radiology, Leiden University Medical Center, Leiden, The Netherlands. L.van_der_Weerd@lumc.nl.
authors.forename: Louise
authors.initials: L
authors.lastname: van der Weerd
authors.affiliation: Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands. S.G.van_Duinen@lumc.nl.
authors.forename: Sjoerd G
authors.initials: SG
authors.lastname: van Duinen
authors.affiliation: Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands. A.M.J.M.van_den_Maagdenberg@lumc.nl.|Department of Neurology, Leiden University Medical Center, Leiden, The Netherlands. A.M.J.M.van_den_Maagdenberg@lumc.nl.
authors.forename: Arn M J M
authors.initials: AM
authors.lastname: van den Maagdenberg
authors.affiliation: Department of Clinical Genetics, K5-R, Leiden University Medical Center, PO Box 9600, 2300, RC, Leiden, The Netherlands. lesnik@lumc.nl.
authors.forename: Saskia A J
authors.initials: SA
authors.lastname: Lesnik Oberstein
mesh_terms: D000367:Age Factors; D000704:Analysis of Variance; D000818:Animals; D001921:Brain / Q000378:metabolism / Q000473:pathology*; D046589:CADASIL / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D004252:DNA Mutational Analysis; D004195:Disease Models, Animal; D005786:Gene Expression Regulation / Q000235:genetics*; D006801:Humans; D008279:Magnetic Resonance Imaging; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008854:Microscopy, Electron; D009154:Mutation / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D000071656:Receptor, Notch3; D051880:Receptors, Notch / Q000235:genetics* / Q000378:metabolism*
chemical_list: C499374:NOTCH3 protein, human; D012333:RNA, Messenger; D000071656:Receptor, Notch3; D051880:Receptors, Notch