Zurich Open Repository and Archive University of Zurich Main Library Strickhofstrasse 39 CH-8057 Zurich www.zora.uzh.ch Year: 2005 Cholesterol 25-hydroxylase on chromosome 10q is a susceptibility gene for sporadic Alzheimer’s disease Papassotiropoulos, A ; Lambert, J C ; Wavrant-De Vrièze, F ; Wollmer, M A ; von der Kammer, H ; Streffer, J R ; Maddalena, A ; Huynh, K D ; Wolleb, S ; Lütjohann, D ; Schneider, B;Thal,DR Grimaldi, L M E ; Tsolaki, M ; Kapaki, E ; Ravid, R ; Konietzko, U ; Hegi, T ; Pasch, T ; Jung, H ; Braak, H ; Amouyel, P ; Rogaev, E I ; Hardy, J ; Hock, C ; Nitsch, R M Abstract: Alzheimer’s disease (AD) is the most common cause of dementia. It is characterized by beta- amyloid (A beta) plaques, neurofibrillary tangles and the degeneration of specifically vulnerable brain neurons. We observed high expression of the cholesterol 25-hydroxylase (CH25H) gene in specifically vulnerable brain regions of AD patients. CH25H maps to a region within 10q23 that has been previ- ously linked to sporadic AD. Sequencing of the 5’ region of CH25H revealed three common haplotypes, CH25Hchi2, CH25Hchi3 and CH25Hchi4; CSF levels of the cholesterol precursor lathosterol were higher in carriers of the CH25Hchi4 haplotype. In 1,282 patients with AD and 1,312 healthy control subjects from five independent populations, a common variation in the vicinity of CH25H was significantly associated with the risk for sporadic AD (p = 0.006). Quantitative neuropathology of brains from elderly non- demented subjects showed brain A beta deposits in carriers of CH25Hchi4 and CH25Hchi3 haplotypes, whereas no A beta deposits were present in CH25Hchi2 carriers. Together, these results are compatible with a role of CH25Hchi4 as a putative susceptibility factor for sporadic AD; they may explain part of the linkage of chromosome 10 markers with sporadic AD, and they suggest the possibility that CH25H polymorphisms are associated with different rates of brain A beta deposition. DOI: https://doi.org/10.1159/000090362 Posted at the Zurich Open Repository and Archive, University of Zurich ZORA URL: https://doi.org/10.5167/uzh-49534 Journal Article Published Version Originally published at: Papassotiropoulos, A; Lambert, J C; Wavrant-De Vrièze, F; Wollmer, M A; von der Kammer, H; Streffer, J R; Maddalena, A; Huynh, K D; Wolleb, S; Lütjohann, D; Schneider, B; Thal, D R; Grimaldi, L M E; Tsolaki, M; Kapaki, E; Ravid, R; Konietzko, U; Hegi, T; Pasch, T; Jung, H; Braak, H; Amouyel, P; Rogaev, E I; Hardy, J; Hock, C; Nitsch, R M (2005). Cholesterol 25-hydroxylase on chromosome 10q is a susceptibility gene for sporadic Alzheimer’s disease. Neurodegenerative Diseases, 2(5):233-241. DOI: https://doi.org/10.1159/000090362 Original Paper Neurodegenerative Dis 2005;2:233–241 Received: March 23, 2005 Diseases DOI: 10.1159/000090362 Accepted after revision: June 6, 2005 Cholesterol 25-Hydroxylase on Chromosome 10q Is a Susceptibility Gene for Sporadic Alzheimer’s Disease a d e Andreas Papassotiropoulos Jean-Charles Lambert Fabienne Wavrant-De Vrièze a g a a M. Axel Wollmer Heinz von der Kammer Johannes R. Streffer Alessia Maddalena a a h h i Kim-Dung Huynh Sibylle Wolleb Dieter Lütjohann Brigitte Schneider Dietmar R. Thal k l m n a Luigi M.E. Grimaldi Magdalini Tsolaki Elisabeth Kapaki Rivka Ravid Uwe Konietzko b b c j d f Thomas Hegi Thomas Pasch Hans Jung Heiko Braak Philippe Amouyel Evgeny I. Rogaev e a a John Hardy Christoph Hock Roger M. Nitsch a b c Division of Psychiatry Research, Institute of Anesthesiology, and Department of Neurology, University of d e Zurich, Zurich , Switzerland; INSERM U508, Institut Pasteur de Lille, Lille , France; Laboratory of Neurogenetics, f National Institute on Aging, National Institutes of Health, Bethesda, Md. , and Department of Psychiatry, Brudnick Neuropsychiatric Research Institute, University of Massachusetts Medical School, Worcester, Mass. , USA; g h i Evotec Neurosciences, Hamburg , Institute of Clinical Pharmacology, University of Bonn, and Institute of j Neuropathology, University of Bonn Medical Center, Bonn , and Institute of Clinical Neuroanatomy, k Johann Wolfgang Goethe University, Frankfurt , Germany; Department of Neuroscience, AUSL n.2, l Caltanissetta , Italy; Third Department of Neurology, Aristotle University of Thessaloniki, Thessaloniki , and m Department of Neurology, ‘Aeginition’ Hospital, University of Athens Medical School, Athens , Greece; n The Netherlands Brain Bank, Amsterdam , The Netherlands Key Words lesterol precursor lathosterol were higher in carriers of Amyloid ؒ Cholesterol ؒ Cholesterol 25-hydroxylase ؒ the CH25H ␹ 4 haplotype. In 1,282 patients with AD and- ␤ Dementia ؒ Genetic association ؒ Lathosterol ؒ 1,312 healthy control subjects from fi ve independent Susceptibility gene ؒ Tau populations, a common variation in the vicinity of CH25H was signifi cantly associated with the risk for sporadic AD (p = 0.006). Quantitative neuropathology of brains from Abstract elderly non-demented subjects showed brain A ␤ depos- Alzheimer’s disease (AD) is the most common cause of its in carriers of CH25H ␹ 4 and CH25H ␹ 3 haplotypes, dementia. It is characterized by ␤ -amyloid (A ␤ ) plaques, whereas no A ␤ deposits were present in CH25H ␹ 2 carri- neurofi brillary tangles and the degeneration of specifi - ers. Together, these results are compatible with a role of cally vulnerable brain neurons. We observed high ex- CH25H ␹ 4 as a putative susceptibility factor for sporadic pression of the cholesterol 25-hydroxylase (CH25H) gene AD; they may explain part of the linkage of chromosome in specifi cally vulnerable brain regions of AD patients. 10 markers with sporadic AD, and they suggest the pos- CH25H maps to a region within 10q23 that has been pre- sibility that CH25H polymorphisms are associated with .viously linked to sporadic AD. Sequencing of the 5 ؅ re- different rates of brain A ␤ deposition gion of CH25H revealed three common haplotypes, Copyright © 2005 S. Karger AG, Basel CH25H ␹ 2, CH25H ␹ 3 and CH25H ␹ 4; CSF levels of the cho- © 2005 S. Karger AG, Basel Andreas Papassotiropoulos 1660–2854/05/0025–0233$22.00/0 Division of Psychiatry Research Fax +41 61 306 12 34 University of Zürich, Lenggstrasse 31 E-Mail [email protected] Accessible online at: CH–8029 Zürich (Switzerland) www.karger.com www.karger.com/ndd Tel. +41 44 384 2644, Fax +41 44 384 2686, E-Mail [email protected] Downloaded by: Universität Zürich, Zentralbibliothek Zürich 130.60.47.22 - 7/7/2016 2:18:28 PM Introduction pocampus and frontal cortex was determined (relative quantifi ca- tion). The following strategy was used for normalization: Xenopus ␤-globin mRNA spiked into the mRNA of each brain tissue was Alzheimer’s disease (AD) is the most common cause used as a qPCR standard to normalize differences in the cDNA of dementia. It is characterized by ␤ -amyloid ( ␤A) plaques, synthesis effi ciency. Additionally, the qPCR procedure was applied neurofi brillary tangles (NFTs) and the degeneration of to a set of housekeeping genes, which were selected as a reference specifi cally vulnerable brain neurons. Degeneration of standard for quantifi cation. The ratio of the amount of frontal and neurons in AD occurs predominantly in such brain re- temporal mRNA of fi ve such housekeeping genes (cyclophilin B, ribosomal protein S9, ␤-actin, GAPDH and transferrin receptor) gions as the hippocampus, the inferior temporal cortex, was determined, the mean value from the fi ve ratios calculated and the entorhinal cortex and the amygdala. The relative pro- used for normalization of candidate gene expression. Primers for tection from degeneration of neuronal populations in the qPCR of CH25H were 5 ؅-GGT CAA CAT CTG GCT TTC CG-3 ؅ -frontal and the occipital cortex [1] indicates selective vul- (forward) and 5 ؅-CAC CAG TCT GTG AGT GGA CCA A-3 ؅ (re nerability of specifi c neurons in brain regions involved in verse). cognitive and memory processes. Genetic Association Studies We previously demonstrated specifi c downregulation Genetic studies were conducted on fi ve independent popula- of seladin-1 (DHCR24), encoding 24-dehydrocholesterol tions: a Swiss sample (174 AD patients and 285 controls), a Medi- reductase (seladin-1), in the inferior temporal cortex in terranean sample from Greece and Italy (272 AD patients and 125 AD using a differential mRNA display approach [2] ; controls), a Russian sample (74 AD patients and 90 controls), a French sample (551 AD patients and 665 controls) and a US sam- downregulation of DHCR24 is reportedly associated with ple derived from the NIMH Human Genetics Initiative and from pathologic phosphorylation of tau, the major protein- the National Cell Repository for Alzheimer’s Disease (NCRAD; aceous constituent of NFTs in AD [3] . grant No. U24 AG21886; 211 AD patients and 147 controls). The In this study, we targeted a priori CH25H, encoding diagnosis of AD was performed according to the NINCDS-ADRDA cholesterol 25-hydroxylase. The intronless CH25H codes criteria. The control groups comprised cognitively healthy elderly individuals who were either the spouses of AD patients or subjects for a polytopic membrane protein of 272 amino acids and recruited from the outpatient clinics of the participating institu- plays an important role in lipid metabolism [4] . By syn- tions. thesizing 25-hydroxycholesterol, a potent co-repressor of SREBP (sterol regulatory element binding protein) pro- Neuropathological Studies cessing, CH25H is involved in the transcriptional regula- Neuropathological examinations were performed in the brains of 71 elderly individuals (mean age of death: 71.6 years, range 50– CH25H tion of lipid-related genes. Importantly, maps 91 years, 28 females) devoid of signifi cant neuropathological ab- within a 30-cM-broad region on chromosome 10q, which normalities and without signs of dementia, as measured by the has been recently linked to late-onset AD [5–7] . In addi- Clinical Dementia Rating scale [15] . The evolutionary phases tion, CH25H is known to be upregulated in the spinal (0–4) of ␤-amyloidosis in the medial temporal lobe of these subjects cord of patients with amyotrophic lateral sclerosis [8] , were determined as described previously [16, 17] .