Platelet Proteomic Analysis Revealed Differential Pattern of Cytoskeletal- and Immune-Related Proteins at Early Stages of Alzheimer’s Disease
Platelets are considered a good model system to study a number of elements associated with neuronal pathways as they share biochemical similarities. Platelets represent the major source of amyloid-β (Aβ) in blood contributing to the Aβ accumulation in the brain parenchyma and vasculature. Peripheral blood platelet alterations including cytoskeletal abnormalities, abnormal cytoplasmic calcium fluxes or increased oxidative stress levels have been related to Alzheimer’s disease (AD) pathology. Therefore, platelets can be considered a peripheral model to study metabolic mechanisms occurring in AD. To investigate peripheral molecular alterations, we examined platelet protein expression in a cohort of 164 subjects, including mild cognitive impairment (MCI), and AD patients, and healthy aged-matched controls. A two-dimensional difference gel electrophoresis (2D-DIGE) discovery phase revealed significant differences between patients and controls in five proteins: talin, vinculin, moesin, complement C3b and Rho GDP, which are known to be involved in cytoskeletal regulation including focal adhesions, inflammation and immune functions. Western blot analysis verified that talin was found to be increased in mild and moderate AD groups versus control, while the other three were found to be decreased. We also analysed amyloid precursor protein (APP), amyloid-β 1-40 (Aβ40) and 1-42 (Aβ42) levels in platelets from the same groups of subjects. Upregulation of platelet APP and Aβ peptides was found in AD patients compared to controls. These findings complement and expand previous reports concerning the morphological and functional alterations in AD platelets, and provide more insights into possible mechanisms that participate in the multifactorial and systemic damage in AD.
KeywordsAlzheimer’s disease Platelets Cytoskeletal proteins Amyloid-β Peripheral changes Amyloid precursor protein
We are grateful to the patients and donors without which this work would not have been possible. We also thank the i+12 Proteomic Unit, in particular Ines Garcia-Consuegra for her advice in the proteomic analysis. We also thank Lola Gutierrez from the UCM CAI-Proteomic Unit helping to analyse and understand the 2D-DIGE results.
Compliance with Ethical Standards
Conflict of Interest
The authors declare they have no conflict of interest.
Ethical Approval and Informed Consent
Blood samples were obtained through antecubital vein puncture. Donors gave written consent, in accordance with the Declaration of Helsinki and the project was approved by the local ethical review committee from the Research Institute Hospital 12 de Octubre (i+12).
- 1.Pellicano M, Bulati M, Buffa S, Barbagallo M, Di Prima A, Misiano G, Picone P, Di Carlo M et al (2010) Systemic immune responses in Alzheimer's disease: in vitro mononuclear cell activation and cytokine production. J Alzheimers Dis 21(1):181–192. https://doi.org/10.3233/jad-2010-091714 CrossRefPubMedGoogle Scholar
- 4.Bartolome F, de Las Cuevas N, Munoz U, Bermejo F, Martin-Requero A (2007) Impaired apoptosis in lymphoblasts from Alzheimer's disease patients: cross-talk of Ca2+/calmodulin and ERK1/2 signaling pathways. Cell Mol Life Sci : CMLS 64(11):1437–1448. https://doi.org/10.1007/s00018-007-7081-3 CrossRefPubMedGoogle Scholar
- 7.Vignini A, Nanetti L, Moroni C, Tanase L, Bartolini M, Luzzi S, Provinciali L, Mazzanti L (2007) Modifications of platelet from Alzheimer disease patients: a possible relation between membrane properties and NO metabolites. Neurobiol Aging 28(7):987–994. https://doi.org/10.1016/j.neurobiolaging.2006.05.010 CrossRefPubMedGoogle Scholar
- 9.Smirnov A, Trupp A, Henkel AW, Bloch E, Reulbach U, Lewczuk P, Riggert J, Kornhuber J et al (2009) Differential processing and secretion of Abeta peptides and sAPPalpha in human platelets is regulated by thrombin and prostaglandine 2. Neurobiol Aging 30(10):1552–1562. https://doi.org/10.1016/j.neurobiolaging.2007.12.009 CrossRefPubMedGoogle Scholar
- 14.Davies TA, Long HJ, Eisenhauer PB, Hastey R, Cribbs DH, Fine RE, Simons ER (2000) Beta amyloid fragments derived from activated platelets deposit in cerebrovascular endothelium: usage of a novel blood brain barrier endothelial cell model system. Amyloid : Int J Exp Clin Investig : Off J Int Soc Amyloidosis 7(3):153–165CrossRefGoogle Scholar
- 15.Kawamoto EM, Munhoz CD, Glezer I, Bahia VS, Caramelli P, Nitrini R, Gorjao R, Curi R et al (2005) Oxidative state in platelets and erythrocytes in aging and Alzheimer's disease. Neurobiol Aging 26(6):857–864. https://doi.org/10.1016/j.neurobiolaging.2004.08.011 CrossRefPubMedGoogle Scholar
- 18.McKhann GM, Knopman DS, Chertkow H, Hyman BT, Jack CR Jr, Kawas CH, Klunk WE, Koroshetz WJ et al (2011) The diagnosis of dementia due to Alzheimer’s disease: recommendations from the National Institute on Aging-Alzheimer’s Association workgroups on diagnostic guidelines for Alzheimer’s disease. Alzheimer’s & Dementia : J Alzheimer's Assoc 7(3):263–269. https://doi.org/10.1016/j.jalz.2011.03.005 CrossRefGoogle Scholar
- 19.Albert MS, DeKosky ST, Dickson D, Dubois B, Feldman HH, Fox NC, Gamst A, Holtzman DM et al (2011) The diagnosis of mild cognitive impairment due to Alzheimer’s disease: recommendations from the National Institute on Aging-Alzheimer’s Association workgroups on diagnostic guidelines for Alzheimer’s disease. Alzheimer’s & Dementia : J Alzheimer’s Assoc 7(3):270–279. https://doi.org/10.1016/j.jalz.2011.03.008 CrossRefGoogle Scholar
- 23.Roher AE, Esh CL, Kokjohn TA, Castano EM, Van Vickle GD, Kalback WM, Patton RL, Luehrs DC et al (2009) Amyloid beta peptides in human plasma and tissues and their significance for Alzheimer’s disease. Alzheimer's Dementia : J Alzheimer's Assoc 5(1):18–29. https://doi.org/10.1016/j.jalz.2008.10.004 CrossRefGoogle Scholar
- 24.Gattaz WF, Talib LL, Schaeffer EL, Diniz BS, Forlenza OV (2014) Low platelet iPLA(2) activity predicts conversion from mild cognitive impairment to Alzheimer’s disease: a 4-year follow-up study. J Neural Transm (Vienna) 121(2):193–200. https://doi.org/10.1007/s00702-013-1088-8 CrossRefGoogle Scholar
- 37.Rogers J, Li R, Mastroeni D, Grover A, Leonard B, Ahern G, Cao P, Kolody H et al (2006) Peripheral clearance of amyloid beta peptide by complement C3-dependent adherence to erythrocytes. Neurobiol Aging 27(12):1733–1739. https://doi.org/10.1016/j.neurobiolaging.2005.09.043 CrossRefPubMedGoogle Scholar
- 39.Vignini A, Morganti S, Salvolini E, Sartini D, Luzzi S, Fiorini R, Provinciali L, Di Primio R et al (2013) Amyloid precursor protein expression is enhanced in human platelets from subjects with Alzheimer’s disease and frontotemporal lobar degeneration: a real-time PCR study. Exp Gerontol 48:1505–1508. https://doi.org/10.1016/j.exger.2013.10.008 CrossRefPubMedGoogle Scholar
- 47.Casoli T, Di Stefano G, Giorgetti B, Balietti M, Recchioni R, Moroni F, Marcheselli F, Bernardini G et al (2008) Platelet as a physiological model to investigate apoptotic mechanisms in Alzheimer beta-amyloid peptide production. Mech Ageing Dev 129(3):154–162. https://doi.org/10.1016/j.mad.2007.11.004 CrossRefPubMedGoogle Scholar
- 49.Johnston JA, Liu WW, Coulson DT, Todd S, Murphy S, Brennan S, Foy CJ, Craig D et al (2008) Platelet beta-secretase activity is increased in Alzheimer’s disease. Neurobiol Aging 29(5):661–668. https://doi.org/10.1016/j.neurobiolaging.2006.11.003 CrossRefPubMedGoogle Scholar
- 51.Donner L, Falker K, Gremer L, Klinker S, Pagani G, Ljungberg LU, Lothmann K, Rizzi F et al (2016) Platelets contribute to amyloid-beta aggregation in cerebral vessels through integrin alphaIIbbeta3-induced outside-in signaling and clusterin release. Sci Signal 9(429):ra52. https://doi.org/10.1126/scisignal.aaf6240 CrossRefPubMedGoogle Scholar
- 53.Wang J, Ohno-Matsui K, Yoshida T, Kojima A, Shimada N, Nakahama K, Safranova O, Iwata N et al (2008) Altered function of factor I caused by amyloid beta: implication for pathogenesis of age-related macular degeneration from Drusen. J Immunol (Baltimore, Md : 1950) 181(1):712–720CrossRefGoogle Scholar