The isoform-specific pathological effects of ApoE4 in vivo are prevented by a fish oil (DHA) diet and are modified by cholesterol

Journal of Alzheimer's Disease

Volumn 28, Issue 3, 2012, Pages 667-683

  Kariv Inbal, Zehavit ^a   Yacobson, Shiri ^a   Berkecz, Robert ^b   Peter, Maria ^c   Janaky, Tamas ^b   Lütjohann, Dieter ^d   Broersen, Laus M ^e   Hartmann, Tobias ^f   Michaelson, Daniel M ^a  

^a TEL AVIV UNIVERSITY   (Israel)

^b UNIVERSITY OF SZEGED   (Hungary)

^c INSTITUTE OF EXPERIMENTAL MEDICINE   (Hungary)

^d UNIVERSITY OF BONN   (Germany)

^e Danone N Research Center for Specialized Nutrition   (Netherlands)

^f SAARLAND UNIVERSITY   (Germany)

Author keywords

Alzheimer's disease; amyloid ; apolipoprotein E4; cholesterol; DHA; GABAergic; glutamatergic

Indexed keywords

AMYLOID BETA PROTEIN[1-42]; APOLIPOPROTEIN E3; APOLIPOPROTEIN E4; CHOLESTEROL; DOCOSAHEXAENOIC ACID; FATTY ACID; FISH OIL; PHOSPHATIDYLCHOLINE; PHOSPHATIDYLETHANOLAMINE; PHOSPHATIDYLSERINE; PHOSPHOLIPID; VESICULAR GLUTAMATE TRANSPORTER; VESICULAR INHIBITORY AMINO ACID TRANSPORTER;

ALZHEIMER DISEASE; ANIMAL EXPERIMENT; ANIMAL MODEL; ARTICLE; BEHAVIOR; CHOLESTEROL METABOLISM; CONTROLLED STUDY; DIETARY INTAKE; ENVIRONMENTAL FACTOR; GENOTYPE; HIPPOCAMPAL CA3 REGION; IN VIVO STUDY; LEARNING; MALE; MEMORY; MOUSE; NEUROPROTECTION; NONHUMAN; PHENOTYPE; PRIORITY JOURNAL; RISK FACTOR; TASK PERFORMANCE;

EID: 84858031544     PISSN: 13872877     EISSN: 18758908     Source Type: Journal    
DOI: 10.3233/JAD-2011-111265     Document Type: Article

References (83)

1. Mahley RW, Rall SC Jr (2000) Apolipoprotein E: Far more than a lipid transport protein. Annu Rev Genomics Hum Genet 1, 507-537.
2. Pitas RE, Boyles JK, Lee SH, Foss D, Mahley RW (1987) Astrocytes synthesize apolipoprotein E and metabolize apolipoprotein E-containing lipoproteins. Biochim Biophys Acta 917, 148-161. (Pubitemid 17231059)
3. Grehan S, Tse E, Taylor JM (2001) Two distal downstream enhancers direct expression of the human apolipoprotein E gene to astrocytes in the brain. J Neurosci 21, 812-822. (Pubitemid 32163122)
4. Aoki K, Uchihara T, Sanjo N, Nakamura A, Ikeda K, Tsuchiya K, Wakayama Y (2003) Increased expression of neuronal apolipoprotein E in human brain with cerebral infarction. Stroke 34, 875-880. (Pubitemid 36402954)
5. Xu PT, Gilbert JR, Qiu HL, Ervin J, Rothrock-Christian TR, Hulette C, Schmechel DE (1999) Specific regional transcription of apolipoprotein E in human brain neurons. Am J Pathol 154, 601-611. (Pubitemid 29094109)
6. Herz J (2009) Apolipoprotein E receptors in the nervous system. Curr Opin Lipidol 20, 190-196.
7. Weisgraber KH (1994) Apolipoprotein E: Structure-function relationships. Adv Protein Chem 45, 249-302.
8. Corder EH, Saunders AM, Strittmatter WJ, Schmechel DE, Gaskell PC, Small GW, Roses AD, Haines JL, Pericak-Vance MA (1993) Gene dose of apolipoprotein E type 4 allele and the risk of Alzheimer's disease in late onset families. Science 261, 921-923. (Pubitemid 23275809)
9. Saunders AM, Strittmatter WJ, Schmechel D, George-Hyslop PH, Pericak-Vance MA, Joo SH, Rosi BL, Gusella JF, Crapper-MacLachlan DR, Alberts MJ et al. (1993) Association of apolipoprotein E allele epsilon 4 with late-onset familial and sporadic Alzheimer's disease. Neurology 43, 1467-1472. (Pubitemid 23230526)
10. Strittmatter WJ, Weisgraber KH, Huang DY, Dong LM, Salvesen GS, Pericak-Vance M, Schmechel D, Saunders AM, Goldgaber D, Roses AD (1993) Binding of human apolipoprotein E to synthetic amyloid beta peptide: Isoformspecific effects and implications for late-onset Alzheimer disease. Proc Natl Acad Sci U S A 90, 8098-8102. (Pubitemid 23260862)
11. Genin E, Hannequin D, Wallon D, Sleegers K, Hiltunen M, Combarros O, Bullido MJ, Engelborghs S, De Deyn P, Berr C, Pasquier F, Dubois B, Tognoni G, Fievet N, Brouwers N, Bettens K, Arosio B, Coto E, Del Zompo M, Mateo I, Epelbaum J, Frank-Garcia A, Helisalmi S, Porcellini E, Pilotto A, Forti P, Ferri R, Scarpini E, Siciliano G, Solfrizzi V, Sorbi S, Spalletta G, Valdivieso F, Vepsalainen S, Alvarez V, Bosco P, Mancuso M, Panza F, Nacmias B, Bossu P, Hanon O, Piccardi P, Annoni G, Seripa D, Galimberti D, Licastro F, Soininen H, Dartigues JF, Kamboh MI, Van Broeckhoven C, Lambert JC, Amouyel P, Campion D (2011) APOE and Alzheimer disease: A major gene with semi-dominant inheritance. Mol Psychiatry 16, 903- 907.
12. MO MC, Muir KW, Weir CJ, Dyker AG, Bone I, Nicoll JA, Lees KR (1998) The apolipoprotein E epsilon4 allele and outcome in cerebrovascular disease. Stroke 29, 1882-1887.
13. Zhou W, Xu D, Peng X, Zhang Q, Jia J, Crutcher KA (2008) Meta-analysis of APOE4 allele and outcome after traumatic brain injury. J Neurotrauma 25, 279-290. (Pubitemid 351457767)
14. Schmechel DE, Saunders AM, Strittmatter WJ, Crain BJ, Hulette CM, Joo SH, Pericak-Vance MA, Goldgaber D, Roses AD(1993) Increased amyloid beta-peptide deposition in cerebral cortex as a consequence of apolipoprotein E genotype in late-onset Alzheimer disease. Proc Natl Acad Sci U S A 90, 9649-9653. (Pubitemid 23315843)
15. Arendt T, Schindler C, Bruckner MK, Eschrich K, Bigl V, Zedlick D, Marcova L (1997) Plastic neuronal remodeling is impaired in patients with Alzheimer's disease carrying apolipoprotein epsilon 4 allele. J Neurosci 17, 516-529. (Pubitemid 27024565)
16. Teter B (2004) ApoE-dependent plasticity in Alzheimer's disease. J Mol Neurosci 23, 167-179.
17. Egensperger R, Kosel S, von Eitzen U, Graeber MB (1998) Microglial activation in Alzheimer disease: Association with APOE genotype. Brain Pathol 8, 439-447. (Pubitemid 28308843)
18. Belinson H, Kariv-Inbal Z, Kayed R, Masliah E, Michaelson DM (2008) Following activation of the amyloid cascade, apolipoprotein E4 drives the in vivo oligomerization of amyloid-beta resulting in neurodegeneration. J Alzheimers Dis 22, 959-970.
19. Belinson H, Lev D, Masliah E, Michaelson DM (2008) Activation of the amyloid cascade in apolipoprotein E4 transgenic mice induces lysosomal activation and neurodegeneration resulting in marked cognitive deficits. J Neurosci 28, 4690- 4701.
20. Belinson H, Michaelson DM (2009) Pathological synergism between amyloid-beta and apolipoprotein E4-the most prevalent yet understudied genetic risk factor for Alzheimer's disease. J Alzheimers Dis 17, 469-481.
21. Hartman RE, Wozniak DF, Nardi A, Olney JW, Sartorius L, Holtzman DM (2001) Behavioral phenotyping of GFAP-apoE3 and -apoE4 transgenic mice: ApoE4 mice show profound working memory impairments in the absence of Alzheimer's-like neuropathology. Exp Neurol 170, 326- 344.
22. Ye S, Huang Y, Mullendorff K, Dong L, Giedt G, Meng EC, Cohen FE, Kuntz ID, Weisgraber KH, Mahley RW (2005) Apolipoprotein (apo) E4 enhances amyloid beta peptide production in cultured neuronal cells: ApoE structure as a potential therapeutic target. Proc Natl Acad Sci U S A 102, 18700-18705. (Pubitemid 43011289)
23. Manelli AM, Bulfinch LC, Sullivan PM, LaDu MJ (2007) Abeta42 neurotoxicity in primary co-cultures: Effect of apoE isoform and Abeta conformation. Neurobiol Aging 28, 1139- 1147.
24. Selkoe DJ (2001) Alzheimer's disease: Genes, proteins, and therapy. Physiol Rev 81, 741-766.
25. Holtzman DM (2004) In vivo effects of ApoE and clusterin on amyloid-beta metabolism and neuropathology. J Mol Neurosci 23, 247-254.
26. Hardy J (2006) Has the amyloid cascade hypothesis for Alzheimer's disease been proved? Curr Alzheimer Res 3, 71-73.
27. Xu Q, Bernardo A, Walker D, Kanegawa T, Mahley RW, Huang Y (2006) Profile and regulation of apolipoprotein E (ApoE) expression in the CNS in mice with targeting of green fluorescent protein gene to the ApoE locus. J Neurosci 26, 4985-4994. (Pubitemid 44315280)
28. Tesseur I, Van Dorpe J, Spittaels K, Van den Haute C, Moechars D, Van Leuven F (2000) Expression of human apolipoprotein E4 in neurons causes hyperphosphorylation of protein tau in the brains of transgenic mice. Am J Pathol 156, 951-964. (Pubitemid 30626953)
29. Brecht WJ, Harris FM, Chang S, Tesseur I, Yu GQ, Xu Q, Dee Fish J, Wyss-Coray T, Buttini M, Mucke L, Mahley RW, Huang Y (2004) Neuron-specific apolipoprotein e4 proteolysis is associated with increased tau phosphorylation in brains of transgenic mice. J Neurosci 24, 2527-2534. (Pubitemid 38327965)
30. Chen Y, Durakoglugil MS, Xian X, Herz J (2010) ApoE4 reduces glutamate receptor function and synaptic plasticity by selectively impairing ApoE receptor recycling. Proc Natl Acad Sci U S A 107, 12011-12016.
31. Rapp A, Gmeiner B, Huttinger M (2006) Implication of apoE isoforms in cholesterol metabolism by primary rat hippocampal neurons and astrocytes. Biochimie 88, 473-483. (Pubitemid 43867898)
32. Michikawa M, Fan QW, Isobe I, Yanagisawa K (2000) Apolipoprotein E exhibits isoform-specific promotion of lipid efflux from astrocytes and neurons in culture. J Neurochem 74, 1008-1016. (Pubitemid 30104296)
33. Sullivan PM, Mezdour H, Aratani Y, Knouff C, Najib J, Reddick RL, Quarfordt SH, Maeda N (1997) Targeted replacement of the mouse apolipoprotein E gene with the common human APOE3 allele enhances diet-induced hypercholesterolemia and atherosclerosis. J Biol Chem 272, 17972-17980. (Pubitemid 27306374)
34. Mihovilovic M, Robinette JB, DeKroon RM, Sullivan PM, Strittmatter WJ (2007) High-fat/high-cholesterol diet promotes a S1P receptor-mediated antiapoptotic activity for VLDL. J Lipid Res 48, 806-815. (Pubitemid 46557885)
35. Jenner AM, Lim WL, Ng MP, Wenk MR, Shui G, Sharman MJ, Gandy SE, Martins RN (2010) The effect of APOE genotype on brain levels of oxysterols in young and old human APOE epsilon2, epsilon3 and epsilon4 knock-in mice. Neuroscience 169, 109-115.
36. Pfrieger FW (2003) Cholesterol homeostasis and function in neurons of the central nervous system. Cell Mol Life Sci 60, 1158-1171.
37. Poirier J, Baccichet A, Dea D, Gauthier S (1993) Cholesterol synthesis and lipoprotein reuptake during synaptic remodelling in hippocampus in adult rats. Neuroscience 55, 81-90. (Pubitemid 23194987)
38. Lutjohann D (2006) Cholesterol metabolism in the brain: Importance of 24S-hydroxylation. Acta Neurol Scand Suppl 185, 33-42.
39. Shobab LA, Hsiung GY, Feldman HH (2005) Cholesterol in Alzheimer's disease. Lancet Neurol 4, 841-852. (Pubitemid 41619439)
40. Stefani M, Liguri G (2009) Cholesterol in Alzheimer's disease: Unresolved questions. Curr Alzheimer Res 6, 15-29.
41. Grosgen S, Grimm MO, Friess P, Hartmann T (2010) Role of amyloid beta in lipid homeostasis. Biochim Biophys Acta 1801, 966-974.
42. Lukiw WJ, Bazan NG (2008) Docosahexaenoic acid and the aging brain. J Nutr 138, 2510-2514.
43. Scott BL, Bazan NG (1989) Membrane docosahexaenoate is supplied to the developing brain and retina by the liver. Proc Natl Acad Sci U S A 86, 2903-2907. (Pubitemid 19112648)
44. Spector AA (2001) Plasma free fatty acid and lipoproteins as sources of polyunsaturated fatty acid for the brain. J Mol Neurosci 16, 159-165; discussion 215-121.
45. Rapoport SI, Rao JS, Igarashi M (2007) Brain metabolism of nutritionally essential polyunsaturated fatty acids depends on both the diet and the liver. Prostaglandins Leukot Essent Fatty Acids 77, 251-261. (Pubitemid 350214520)
46. Cole GM, Ma QL, Frautschy SA (2009) Omega-3 fatty acids and dementia. Prostaglandins Leukot Essent Fatty Acids 81, 213-221.
47. Oksman M, Iivonen H, Hogyes E, Amtul Z, Penke B, Leenders I, Broersen L, Lutjohann D, Hartmann T, Tanila H (2006) Impact of different saturated fatty acid, polyunsaturated fatty acid and cholesterol containing diets on beta-amyloid accumulation in APP/PS1 transgenic mice. Neurobiol Dis 23, 563-572. (Pubitemid 44234092)
48. Jicha GA, Markesbery WR (2010) Omega-3 fatty acids: Potential role in the management of early Alzheimer's disease. Clin Interv Aging 5, 45-61.
49. Levi O, Jongen-Relo AL, Feldon J, Roses AD, Michaelson DM (2003) ApoE4 impairs hippocampal plasticity isoform specifically and blocks the environmental stimulation of synaptogenesis and memory. Neurobiol Dis 13, 273-282. (Pubitemid 36937209)
50. Levi O, Michaelson DM (2007) Environmental enrichment stimulates neurogenesis in apolipoprotein E3 and neuronal apoptosis in apolipoprotein E4 transgenic mice. J Neurochem 100, 202-210. (Pubitemid 46085849)
51. Matyash V, Liebisch G, Kurzchalia TV, Shevchenko A, Schwudke D (2008) Lipid extraction by methyl-tert-butyl ether for high-throughput lipidomics. J Lipid Res 49, 1137- 1146.
52. Balogh G, Peter M, Liebisch G, Horvath I, Torok Z, Nagy E, Maslyanko A, Benko S, Schmitz G, Harwood JL, Vigh L (2010) Lipidomics reveals membrane lipid remodelling and release of potential lipid mediators during early stress responses in a murine melanoma cell line. Biochim Biophys Acta 1801, 1036-1047.
53. Lutjohann D, Brzezinka A, Barth E, Abramowski D, Staufenbiel M, von Bergmann K, Beyreuther K, Multhaup G, Bayer TA (2002) Profile of cholesterol-related sterols in aged amyloid precursor protein transgenic mouse brain. J Lipid Res 43, 1078-1085. (Pubitemid 34760879)
54. Bales KR, Liu F, Wu S, Lin S, Koger D, DeLong C, Hansen JC, Sullivan PM, Paul SM (2009) Human APOE isoformdependent effects on brain beta-amyloid levels in PDAPP transgenic mice. J Neurosci 29, 6771-6779.
55. Ennaceur A (2010) One-trial object recognition in rats and mice: Methodological and theoretical issues. Behav Brain Res 215, 244-254.
56. Wassall SR, Stillwell W (2009) Polyunsaturated fatty acidcholesterol interactions: Domain formation in membranes. Biochim Biophys Acta 1788, 24-32.
57. Du C, Sato A, Watanabe S, Wu CZ, Ikemoto A, Ando K, Kikugawa K, Fujii Y, Okuyama H (2003) Cholesterol synthesis in mice is suppressed but lipofuscin formation is not affected by long-term feeding of n-3 fatty acid-enriched oils compared with lard and n-6 fatty acid-enriched oils. Biol Pharm Bull 26, 766-770. (Pubitemid 39655292)
58. Guo M, Stockert L, Akbar M, Kim HY (2007) Neuronal specific increase of phosphatidylserine by docosahexaenoic acid. J Mol Neurosci 33, 67-73. (Pubitemid 47574960)
59. Corsini A, Verri D, Raiteri M, Quarato P, Paoletti R, Fumagalli R (1995) Effects of 26-aminocholesterol, 27-hydroxycholesterol, and 25-hydroxycholesterol on proliferation and cholesterol homeostasis in arterial myocytes. Arterioscler Thromb Vasc Biol 15, 420-428.
60. Leoni V, Masterman T, Patel P, Meaney S, Diczfalusy U, Bjorkhem I (2003) Side chain oxidized oxysterols in cerebrospinal fluid and the integrity of blood-brain and bloodcerebrospinal fluid barriers. J Lipid Res 44, 793-799. (Pubitemid 37279681)
61. Grimm MO, Kuchenbecker J, Grosgen S, Burg VK, Hundsdorfer B, Rothhaar TL, Friess P, de Wilde MC, Broersen LM, Penke B, Peter M, Vigh L, Grimm HS, Hartmann T (2011) Docosahexaenoic acid reduces amyloid {beta} production via multiple pleiotropic mechanisms. J Biol Chem 286, 14028-14039.
62. Ma QL, Teter B, Ubeda OJ, Morihara T, Dhoot D, Nyby MD, Tuck ML, Frautschy SA, Cole GM (2007) Omega-3 fatty acid docosahexaenoic acid increases SorLA/LR11, a sorting protein with reduced expression in sporadic Alzheimer's disease (AD): Relevance to AD prevention. J Neurosci 27, 14299- 14307.
63. Du J, Zhang L, Liu S, Wang Z (2010) Palmitic acid and docosahexaenoic acid opposingly regulate the expression of insulin-degrading enzyme in neurons. Pharmazie 65, 231- 232.
64. Cirrito JR, Yamada KA, Finn MB, Sloviter RS, Bales KR, May PC, Schoepp DD, Paul SM, Mennerick S, Holtzman DM (2005) Synaptic activity regulates interstitial fluid amyloidbeta levels in vivo. Neuron 48, 913-922. (Pubitemid 41814696)
65. Grimm MO, Grimm HS, Hartmann T (2007) Amyloid beta as a regulator of lipid homeostasis. Trends Mol Med 13, 337- 344.
66. Grimm MO, Grimm HS, Patzold AJ, Zinser EG, Halonen R, Duering M, Tschape JA, De Strooper B, Muller U, Shen J, Hartmann T (2005) Regulation of cholesterol and sphingomyelin metabolism by amyloid-beta and presenilin. Nat Cell Biol 7, 1118-1123.
67. Dyall SC, Michael-Titus AT (2008) Neurological benefits of omega-3 fatty acids. Neuromolecular Med 10, 219-235.
68. Zhong N, Weisgraber KH (2009) Understanding the basis for the association of apoE4 with Alzheimer's disease: Opening the door for therapeutic approaches. Curr Alzheimer Res 6, 415-418.
69. Cao D, Kevala K, Kim J, Moon HS, Jun SB, Lovinger D, Kim HY (2009) Docosahexaenoic acid promotes hippocampal neuronal development and synaptic function. J Neurochem 111, 510-521.
70. Quinn JF, Raman R, Thomas RG, Yurko-Mauro K, Nelson EB, Van Dyck C, Galvin JE, Emond J, Jack CR Jr, Weiner M, Shinto L, Aisen PS (2010) Docosahexaenoic acid supplementation and cognitive decline in Alzheimer disease: A randomized trial. JAMA 304, 1903-1911.
71. Laurin D, Verreault R, Lindsay J,Dewailly E, Holub BJ (2003) Omega-3 fatty acids and risk of cognitive impairment and dementia. J Alzheimers Dis 5, 315-322. (Pubitemid 37411779)
72. Barberger-Gateau P, Raffaitin C, Letenneur L, Berr C, Tzourio C, Dartigues JF, Alperovitch A (2007) Dietary patterns and risk of dementia: The Three-City cohort study. Neurology 69, 1921-1930. (Pubitemid 350100350)
73. Morris MC, Evans DA, Bienias JL, Tangney CC, Bennett DA, Wilson RS, Aggarwal N, Schneider J (2003) Consumption of fish and n-3 fatty acids and risk of incident Alzheimer disease. Arch Neurol 60, 940-946. (Pubitemid 36858352)
74. Huang TL, Zandi PP, Tucker KL, Fitzpatrick AL, Kuller LH, Fried LP, Burke GL, Carlson MC (2005) Benefits of fatty fish on dementia risk are stronger for those without APOE epsilon4. Neurology 65, 1409-1414. (Pubitemid 41552771)
75. Schaefer EJ, Bongard V, Beiser AS, Lamon-Fava S, Robins SJ, Au R, Tucker KL, Kyle DJ, Wilson PW, Wolf PA (2006) Plasma phosphatidylcholine docosahexaenoic acid content and risk of dementia and Alzheimer disease: The Framingham Heart Study. Arch Neurol 63, 1545-1550. (Pubitemid 44748796)
76. Whalley LJ, Deary IJ, Starr JM, Wahle KW, Rance KA, Bourne VJ, Fox HC (2008) n-3 Fatty acid erythrocyte membrane content, APOE varepsilon4, and cognitive variation: An observational follow-up study in late adulthood. Am J Clin Nutr 87, 449-454. (Pubitemid 351226084)
77. Dietschy JM, Turley SD (2001) Cholesterol metabolism in the brain. Curr Opin Lipidol 12, 105-112. (Pubitemid 32242918)
78. Perez SE, Berg BM, Moore KA, He B, Counts SE, Fritz JJ, Hu YS, Lazarov O, Lah JJ, Mufson EJ (2010) DHA diet reduces AD pathology in young APPswe/PS1 Delta E9 transgenic mice: Possible gender effects. J Neurosci Res 88, 1026-1040.
79. Arendash GW, Jensen MT, Salem N Jr, Hussein N, Cracchiolo J, Dickson A, Leighty R, Potter H (2007) A diet high in omega-3 fatty acids does not improve or protect cognitive performance in Alzheimer's transgenic mice. Neuroscience 149, 286-302. (Pubitemid 47576542)
80. Hooijmans CR, Van der Zee CE, Dederen PJ, Brouwer KM, Reijmer YD, van Groen T, Broersen LM, Lutjohann D, Heerschap A, Kiliaan AJ (2009) DHA and cholesterol containing diets influence Alzheimer-like pathology, cognition and cerebral vasculature in APPswe/PS1dE9 mice. Neurobiol Dis 33, 482-498.
81. Caselli RJ, Dueck AC, Osborne D, Sabbagh MN, Connor DJ, Ahern GL, Baxter LC, Rapcsak SZ, Shi J, Woodruff BK, Locke DE, Snyder CH, Alexander GE, Rademakers R, Reiman EM (2009) Longitudinal modeling of age-related memory decline and the APOE epsilon4 effect. N Engl J Med 361, 255-263.
82. Filippini N, MacIntosh BJ, Hough MG, Goodwin GM, Frisoni GB, Smith SM, Matthews PM, Beckmann CF, Mackay CE (2009) Distinct patterns of brain activity in young carriers of the APOE-epsilon4 allele. Proc Natl Acad Sci U S A 106, 7209-7214.
83. Salloway S, Sperling R, Gilman S, Fox NC, Blennow K, Raskind M, Sabbagh M, Honig LS, Doody R, van Dyck CH, Mulnard R, Barakos J, Gregg KM, Liu E, Lieberburg I, Schenk D, Black R, Grundman M (2009) A phase 2 multiple ascending dose trial of bapineuzumab in mild to moderate Alzheimer disease. Neurology 73, 2061-2070.