1.
Structural Basis of C-terminal β-Amyloid Peptide Binding by the AntibodyPonezumab for the Treatment of Alzheimer's Disease.
Source
Rinat, Pfizer Inc., 230 East Grand Avenue, South San Francisco, CA 94080, USA.
Abstract
Alzheimer's disease, the most common cause of dementia in the elderly and characterized by the deposition and accumulation of plaques, is composed in part of β-amyloid (Aβ) peptides, loss of neurons, and the accumulation of neurofibrillary tangles. Here, we describe ponezumab, a humanized monoclonal antibody, and show how it binds specifically to the carboxyl (C)-terminus of Aβ40. Ponezumab can label Aβ that is deposited in brain parenchyma found in sections from Alzheimer's disease casualties and in transgenic mouse models that overexpress Aβ. Importantly, ponezumab does not label full-length, non-cleaved amyloid precursor protein on the cell surface. The C-terminal epitope of ponezumab appears to be available for binding soluble Aβ present in the circulation because systemic administration of ponezumab greatly elevates plasma Aβ40 levels in a dose-dependent fashion after administration to a mouse model that overexpress human Aβ. Administration of ponezumab to transgenic mice also led to a dose-dependent reduction in hippocampal amyloid load. To further explore the nature of ponezumab binding to Aβ40, we determined the X-ray crystal structure of ponezumab in complex with Aβ40 and found that the Aβ40 carboxyl moiety makes extensive contacts with ponezumab. Furthermore, the structure-function analysis supported this critical requirement for carboxy group of AβV40 in the Aβ-ponezumab interaction. These findings provide novel structural insights into the in vivo conformation of the C-terminus of Aβ40 and the brain Aβ-lowering efficacy that we observed following administration of ponezumab in transgenic mouse models.
Copyright © 2011 Elsevier Ltd. All rights reserved.
Molecular Imaging of Amyloidosis: Will the Heart Be the Next Target After the Brain?
Source
Department of Diagnostic Radiology and Nuclear Medicine, University of Maryland School of Medicine, 22 South Greene Street, Baltimore, MD, 21201, USA, wchen5@umm.edu.
Abstract
Amyloidosis is a heterogeneous group of diseases with a common feature of extracellular deposition and infiltration of different types of amyloid fibrils in various organs. For example, Alzheimer's disease is characterized by deposition ofamyloid β in the brain. Radiolabeled positron emission tomography (PET) tracers, mainly derivatives of thioflavin-T, were recently introduced for identification of amyloid β plaques in Alzheimer's patients. Such advances of amyloid β plaque imaging of the brain may shed light into imaging of other organs in amyloidosis patients, such as the heart. Cardiac infiltration of amyloid confers poor clinical outcomes, which renders early diagnosis for appropriate clinical management. At present, nuclear imaging of cardiac amyloidosis is predominantly accomplished with bone-seeking radiotracers, such as 99m-technetium-labeled pyrophosphate ((99m)Tc-PYP), 99m-technetium-methylene diphosphonate ((99m)Tc-MDP), and 99m-technetium-3,3,-diphosphono-1,2-propanodicarboxylic acid ((99m)Tc-DPD), with conflicting results in terms of diagnostic performance, with the exception for (99m)Tc-DPD, which may differentiate light-chain amyloidosis from transthyretin-related cardiac amyloidosis. Although other non-bone-seeking radiotracers such as iodine-123-labeledamyloid P component ((123)I-SAP), 123-iodine-Meta-iodobenzylguanidine ((123)I-mIBG), 99m-technetium-labeled protease inhibitor, and indium-111-labeled amyloid antibodies have also shown some success in identifying cardiac amyloidosis, the future, however, may lie in labeling derivatives of thioflavin-T. With the recent success of visualizing deposition of amyloid β in the brain, the US Food and Drug Administration-approved PET imaging agent (18)F-florbetapir may be used to target cardiac amyloidosis next.
Autoreactive-Aβ antibodies promote APP β-secretase processing.
Source
Rashid Laboratory for Developmental Neurobiology, Department of Psychiatry and Neurosciences, College of medicine, University of South Florida, Tampa, FL, USA Department of Neurology, Daping Hospital, Third Military Medical University, Chongqing, China Neuroimmunology Laboratory, Department of Psychiatry and Neurosciences, College of medicine, University of South Florida, Tampa, FL, USA Departments of Biomedical Sciences and Pathology, Saitama Medical Center and Saitama Medical University, Kawagoe, Saitama, Japan Department of Neuropathology, Heinrich-Heine-University, Düsseldorf, Germany Department of Demyelinating Disease and Aging, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Kodaira, Tokyo, Japan James A. Haley Veterans' Administration Hospital, Tampa, FL, USA.
Abstract
Several prior investigations of Alzheimer's disease (AD) patients have indicated naturally-occurring autoantibodies against amyloid-β (Aβ) species are produced. While many studies have focused on the relative concentrations or binding affinities of autoantibodies against Aβ-related proteins in AD and aging, data regarding their functional properties are limited. It is generally believed that these antibodies act to aid in clearance of Aβ. However, as antibodies which bind to Aβ also typically bind to the parent amyloid precursor protein (APP), we reasoned that certain Aβ-targeting autoantibodies may bind to APP thereby altering its conformation and processing. Here we show for the first time, that naturally occurring Aβ-reactive autoantibodies isolated from AD patients, but not from healthy controls, promote β-secretase activity in cultured cells. Further, using monoclonal antibodies to various regions of Aβ, we found thatantibodies generated against the N-terminal region, especially Aβ(1-17) , dose dependently promoted amyloidogenic processing of APP viaβ-secretase activation. Thus, this property of certain autoantibodies in driving Aβ generation could be of etiological importance in the development of sporadic forms of AD. Furthermore, future passive or active anti-Aβ immunotherapies must consider potential off-target effects resulting from antibodies targeting the N-terminus of Aβ, as co-binding to the corresponding region of APP may actually enhance Aβ generation. © 2011 The Authors Journal of Neurochemistry© 2011 International Society for Neurochemistry.
© 2011 The Authors Journal of Neurochemistry © 2011 International Society for Neurochemistry.
p75NTR is mainly responsible for Aβ toxicity but not for its internalization: a primary study.
Source
Department of Human Physiology and Centre for Neuroscience, Flinders University, GPO Box 2100, Adelaide, 5001, Australia, yuhlzjl@ccmu.edu.cn.
Abstract
Accumulating evidence indicates that the intraneuronal accumulation of beta-amyloid peptide (Aβ) is earlier than the formation of extraneuronal amyloid plaque but the mechanism of the accumulation remains unclear. p75NTR is a receptor for Aβ and interacts with Aβ in vitro and in vivo but whether p75NTR mediates Aβ internalization and intraneuronal accumulation is not known. In this study, we aim to determine if p75NTR mediates Aβ internalization, which might provide new insights into Aβ metabolism and toxicity. FRET analysis in PC12 cells showed that internalized Aβ was close to p75NTR. Aβ1-42 could be internalized in PC12 cells in a concentration-dependent manner but the antibody to the p75NTR extracellular domain did not prevent its internalization. Aβ1-42 could also be internalized in mouse neonatal cortical neurons and the deletion of p75NTR in these neurons did not prevent its internalization but prevented Aβ neurotoxicity. Cholesterol at 10 μM significantly increased Aβ1-42 internalization in PC12 cells. Internalized Aβ1-42 is mainly co-localized with Beclin-1 (a biomarker of autophagosomes) but not with endosomal and lysomal markers. p75NTR may not play a main role in Aβ internalization at the concentrations tested but is responsible for Aβ induced toxicity in primary neurons. Internalized Aβ is mainly sorted to autophagosomes for metabolism.
125I-Labeled single-chain monoclonal antibody, NS4F5, that targets the GlcNS6S-IdoA2S motif of heparan sulfate proteoglycans for the in vivo imaging of peripheral amyloidosis.
Authors
Source
Molecular Imaging and Contrast Agent Database (MICAD) [Internet]. Bethesda (MD): National Center for Biotechnology Information (US); 2004-2011.
2011 Nov 04 [updated 2011 Dec 01].
Excerpt
Peripheral amyloidosis is the extracellular deposition of insoluble protein fibrils in various organs of animals, including humans, and these deposits are considered to be biomarkers for diseases such as Alzheimer’s disease (AD), light chain amyloidosis (AL), etc (1). The fibrils are made up of disease-specific aggregated proteins or peptides (e.g., Aβ peptides for AD, light chains for AL or multiple myeloma, and reactive amyloidosis (AA)) that incorporate heparin sulfate proteoglycans (HSPG; heparin belongs to the heparan sulfate family of proteoglycans that contain the (GlcNS6S-IdoA2S)3 motif)) and the serum amyloid P component (SAP) within their structure during disease progression. A characteristic feature of the protein fibrils is that the constituent proteins form a secondary cross-β pleated sheet structure that is resistant to proteolytic digestion (for structural details, see Goldsbury et al. (2)). In addition, clinical symptoms of amyloidosis in a patient are influenced by the degree to which an organ is involved in the disease (3).The HSPG contain diverse types of oligosaccharides that are sulfated on the hydroxyl moieties to varying degrees, and these hypersulfated structures are distinct, are found specifically in the amyloid deposits, and differ from one another depending on the organ where they are located (4). In addition, the HSPG contain stretches of the disaccharide GlcNS6S-IdoA2S (HSNS4F5; these motifs are abundantly found in heparin, but have limited presence in the HS of normal tissues) that inhibit cell proliferation and induce apoptosis but do not affect the attachment of cells to collagen I (5). Because the HSPG in the amyloids are hypersulfated compared to the proteoglycans in normal tissues, HSPG are considered to be suitable for the detection of amyloids with imaging techniques and to diagnose and monitor amyloidosis progression and to determine the prognosis for a patient with amyloidosis (1). Whole-body scintigraphy with radioiodinated SAP is commonly used in Europe for the detection of amyloidosis in the various parts of the body, but this technique is not approved by the United States Food and Drug Administration for clinical use in United States because the SAP used to generate the tracer is isolated from human sources (1). In an attempt to develop an amyloidimaging agent that would not require the use of human materials, Wall et al. showed that a 125I-labeled synthetic heparin-binding peptide, p5, could be used with single-photon emission computed tomography (SPECT) to detectamyloid deposits in H2/huIL-6 transgenic mice with severe systemic AA amyloidosis (AA mice) (1). In another study, Wall et al. evaluated the use of 125I-labeled single chain (scFv) antibodies (Abs) directed against HS for the detection of amyloids in AA mice that are prone to develop amyloidosis at ~4–5 months of age as described elsewhere (6). Among these radioiodinated scFv Abs, NS4F5, which binds specifically to HSNS4F5 (5), has been determined to be the most suitable for the noninvasive imaging of amyloids in the transgenic animals (6).
Sections
Structure-based design of conformation- and sequence-specific antibodiesagainst amyloid β
Source
Center for Biotechnology and Interdisciplinary Studies, Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180.
Abstract
Conformation-specific antibodies that recognize aggregated proteins associated with several conformational disorders (e.g., Parkinson and prion diseases) are invaluable for diagnostic and therapeutic applications. However, no systematic strategy exists for generating conformation-specific antibodies that target linear sequence epitopes within misfolded proteins. Here we report a strategy for designing conformation- and sequence-specific antibodies against misfolded proteins that is inspired by the molecular interactions governing protein aggregation. We find that grafting small amyloidogenic peptides (6-10 residues) from the Aβ42 peptide associated with Alzheimer's disease into the complementarity determining regions of a domain (V(H)) antibody generates antibody variants that recognize Aβ soluble oligomers and amyloid fibrils with nanomolar affinity. We refer to these antibodies as gammabodies for grafted amyloid-motif antibodies. Gammabodies displaying the central amyloidogenic Aβ motif () are reactive with Aβ fibrils, whereas those displaying the amyloidogenic C terminus () are reactive with Aβ fibrils and oligomers (and weakly reactive with Aβ monomers). Importantly, we find that the grafted motifs target the corresponding peptide segments within misfolded Aβ conformers. Aβ gammabodies fail to cross-react with other amyloidogenic proteins and scrambling their grafted sequences eliminates antibody reactivity. Finally, gammabodies that recognize Aβ soluble oligomers and fibrils also neutralize the toxicity of each Aβ conformer. We expect that our antibody design strategy is not limited to Aβ and can be used to readily generate gammabodies against other toxic misfolded proteins.
E. coli chaperones DnaK, Hsp33 and Spy inhibit bacterial functional amyloidassembly.
Source
Department of Molecular, Cellular and Developmental Biology; University of Michigan, Ann Arbor, MI USA.
Abstract
Amyloid formation is an ordered aggregation process, where β-sheet rich polymers are assembled from unstructured or partially folded monomers. We examined how two Escherichia coli cytosolic chaperones, DnaK and Hsp33, and a more recently characterized periplasmic chaperone, Spy, modulate the aggregation of a functional amyloid protein, CsgA. We found that DnaK, the Hsp70 homologue in E. coli, and Hsp33, a redox-regulated holdase, potently inhibited CsgA amyloidogenesis. The Hsp33 anti-amyloidogenesis activity was oxidation dependent, as oxidized Hsp33 was significantly more efficient than reduced Hsp33 at preventing CsgA aggregation. When soluble CsgA was seeded with preformed amyloid fibers, neither Hsp33 nor DnaK were able to efficiently prevent soluble CsgA from adopting theamyloid conformation. Moreover, both DnaK and Hsp33 increased the time that CsgA was reactive with the amyloidoligomer conformation-specific A11 antibody. Since CsgA must also pass through the periplasm during secretion, we assessed the ability of the periplasmic chaperone Spy to inhibit CsgA polymerization. Like DnaK and Hsp33, Spy also inhibited CsgA polymerization in vitro. Overexpression of Spy resulted in increased chaperone activity in periplasmic extracts and in reduced curli biogenesis in vivo. We propose that DnaK, Hsp33 and Spy exert their effects during the nucleation stages of CsgA fibrillation. Thus, both housekeeping and stress induced cytosolic and periplasmic chaperones may be involved in discouraging premature CsgA interactions during curli biogenesis.
Bispecific Tandem Single Chain Antibody Simultaneously Inhibits β-Secretase and Promotes α-Secretase Processing of AβPP.
Source
Department of Chemical Engineering, Arizona State University, Tempe, Arizona, AZ, USA.
Abstract
Misfolding and aggregation of amyloid-β (Aβ) is an important early event in the pathogenesis of Alzheimer's disease. Aβ is produced by sequential proteolysis of the amyloid-β protein precursor (AβPP) by β- and γ-secretases. A third protease, α-secretase, cleaves AβPP in the middle of the Aβ sequence precluding formation of Aβ. The levels of Aβ generated from AβPP can therefore be controlled by tailoring activity of these proteases toward AβPP. We previously showed that β-secretase proteolysis of AβPP could be selectively inhibited using the single chain antibody fragment (scFv) iBSEC1, which blocks the cleavage site on AβPP, and α-secretase proteolysis of AβPP could be selectively enhanced using a proteolytic scFv (Asec1A) engineered to have α-secretase-like activity. Here we show that DIA10D, a novel tandem bispecific scFv combining iBSEC1 with the ASec1A can control amyloidogenic processing of AβPP by simultaneously inhibiting β-secretase and increasing α-secretase processing of AβPP. When expressed in H4 (neuroglioma) cells overexpressing AβPP, DIA10D potently reduces levels of extracellular Aβ by around 50% while also increasing levels of neuroprotective sAβPPα. DIA10D activity has been designed to selectively target AβPP, so this modulation of AβPP processing should not affect endogenous activity of α-and β-secretases towards other substrates.
Amyloid precursor protein revisited: Neuronal-specific expression and the highly stable nature of soluble derivatives.
Source
Baylor College of Medicine, United States.
Abstract
APP processing and Aβ production plays a central role in Alzheimer disease (AD) pathogenesis. APP has been considered a ubiquitously expressed protein. In addition to Aβ, α- or β-secretase dependent cleavage of APP also generates soluble secreted APP (APPsα or APPsβ respectively). Interestingly, APPsβ has been shown to be subject to further cleavage to create an N-APP fragment which binds to the DR6 death receptor and mediates axon pruning and degeneration under trophic factor withdrawal conditions. By performing APP immunocytochemical staining, we found that, unexpectedly, many antibodies yielded non-specific staining in APP null samples. Screening of a series ofantibodies allowed us to identify a rabbit monoclonal antibody Y188 that is highly specific for APP and prompted us to re-examine the expression, localization and stability of endogenous APP and APPsβ in wild-type and in APPsβ knock-in mice, respectively. In contrast to earlier reports, we found that APP is specifically expressed in neurons and its expression cannot be detected in major types of glial cells under basal or neuroinflammatory conditions. Both APPsα and APPsβ are highly stable in the central nervous system (CNS) and do not undergo further cleavage with or without trophic factor support. Our results clarify several key questions with regards to the fundamental properties of APP and offer critical cellular insights into the pathophysiology of APP.
The senescence accelerated mouse (SAMP8) as a model for oxidative stress and Alzheimer's disease.
Source
Division of Geriatric Medicine, Saint Louis University School of Medicine, St. Louis, MO, USA.
Abstract
The senescence accelerated mouse (SAMP8) is a spontaneous animal model of overproduction of amyloid precursor protein (APP) and oxidative damage. It develops early memory disturbances and changes in the blood-brain barrier resulting in decreased efflux of amyloid-β protein from the brain. It has a marked increase in oxidative stress in the brain. Pharmacological treatments that reduce oxidative stress improve memory. Treatments that reduce amyloid-β (antisense to APP and antibodies to amyloid-β) not only improve memory but reduce oxidative stress. Early changes in lipid peroxidative damage favor mitochondrial dysfunction as being a trigger for amyloid-β overproduction in this genetically susceptible mouse strain. This sets in motion a cycle where the increased amyloid-beta further damages mitochondria. We suggest that this should be termed the Inflammatory-Amyloid Cycle and may well be similar to the mechanisms responsible for the pathophysiology of Alzheimer's disease. This article is part of a Special Issue entitled: Antioxidants and Antioxidant Treatment in Disease.
Copyright © 2011. Published by Elsevier B.V.
Comparison of Pharmacokinetics, Pharmacodynamics, Safety, and Tolerability of the Amyloid β Monoclonal Antibody Solanezumab in Japanese and White Patients With Mild to Moderate Alzheimer Disease.
Source
*Lilly Research Laboratories Japan, Eli Lilly Japan K.K., Kobe; †Department of Clinical Pharmacokinetics, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan; and ‡Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN.
Abstract
OBJECTIVES:
Solanezumab is a humanized anti-amyloid β monoclonal antibody being developed as a passive immunization treatment to slow the progression of Alzheimer disease (AD). Pharmacokinetics (PK), pharmacodynamics, safety, and tolerability after a single dose of solanezumab were compared between Japanese and white patients with AD.
METHODS:
Japanese and white patients with mild to moderate AD were enrolled in 2 separate studies. In each study, single doses of solanezumab at 0.5, 1.5, 4.0, and 10.0 mg/kg were administered by intravenous infusion. Plasma concentrations of solanezumab and amyloid β (Aβ) were measured. A safety assessment was conducted up to 112 days after a single-dose administration of solanezumab.
RESULTS:
The PK profile was similar between the Japanese and the white patients with AD. In both the Japanese and the white patients, clearance and volume of distribution appeared similar across doses, suggesting that solanezumab exhibited dose-proportional PK within the studied dose range. A marked increase in plasma total Aβ was observed; both the magnitude and time to reach maximum concentration tended to increase with increasing doses of solanezumab. Administration of solanezumab was generally well-tolerated in both Japanese and white patients with AD.
CONCLUSIONS:
When administered as a per-kilogram single dose of solanezumab, PK and pharmacodynamics (plasma total Aβ1-40 concentration) in the Japanese patients with AD were comparable with those in the white patients with AD. In addition, solanezumab was generally well tolerated in both Japanese and white patients at all dose levels.
The Arctic amyloid-β precursor protein (AβPP) mutation results in distinct plaques and accumulation of N- and C-truncated Aβ
Source
Department of Public Health and Caring Sciences, Uppsala University, Uppsala, Sweden.
Abstract
The Arctic (p. E693G) mutation in the amyloid-β precursor protein (AβPP) facilitates amyloid-β (Aβ) protofibril formation and generates clinical symptoms of Alzheimer's disease (AD). Here, molecular details of Aβ in post mortem brain were investigated with biochemical and morphological techniques. The basic structure of Arctic plaques resembled cotton wool plaques. However, they appeared ring-formed with Aβ42-specific antibodies, but were actually targetoid, since the periphery and center of many parenchymal Aβ deposits stained differently with mid-domain, N- and C-terminal Aβantibodies. Aβ fibrils were similar in shape, albeit shorter than in sporadic AD brain, when examined by electron microscopy. Aβwild-type and Aβarctic codeposited and parenchymal deposits were highly enriched in both N- and C-terminally truncated Aβ. In contrast, cerebral amyloid angiopathy (CAA) contained a substantial amount of Aβ1-40. The absence of plaques with cores of fibrillary Aβ might be due to the scarcity of full-length Aβ, although other mechanisms could be involved. Our findings are discussed in relation to mechanisms and relevance of amyloid formation and to the clinical features of AD.
Copyright © 2011 Elsevier Inc. All rights reserved.
AβPP Accumulation and/or Intraneuronal Amyloid-β Accumulation? The 3xTg-AD Mouse Model Revisited.
Source
Division of Molecular Psychiatry, Georg-August-University Göttingen, University Medicine Göttingen, Göttingen, Germany.
Abstract
The triple-transgenic Alzheimer's disease (AD) mouse model, 3xTg-AD, played an important role in supporting the intraneuronal amyloid-β (Aβ) hypothesis in AD. However, recent evidence claims that the 3xTg-AD mice accumulateamyloid-β protein precursor (AβPP) instead of Aβ within neurons. In the present report, we re-investigated the 3xTg-AD mouse model and confirmed recent findings of age-dependent AβPP accumulations. In addition, intraneuronal Aβ was detected mainly in the neocortex using conformation-specific as well as antibodies directed against Aβ neo-epitopes. In contrast to previous work, however, only minor levels of intraneuronal Aβ were detected in the CA1 region of the hippocampus in aged 3xTg-AD mice.
Diagnostic value of markers of muscle degeneration in sporadic inclusion body myositis.
Source
Laboratoire de Neuropathologie, Institut de Myologie, Assistance Publique, Hôpitaux de Paris, Hôpital Pitié-Salpêtrière, Université Pierre et Marie Curie, Paris, France. odile.dubourg@psl.aphp.fr
Abstract
Sporadic inclusion body myositis (s-IBM) is characterized histologically by the association of concomitant inflammatory and degenerative processes. We evaluated the sensitivity and specificity of different markers of the degenerative process in order to refine the histological diagnosis. We performed an immunohistochemical study with antibodies directed against ubiquitin, amyloid-beta precursor protein (AbetaPP), amyloid-beta (Abeta), SMI-31, SMI-310, Tar-DNA binding protein-43 (TDP-43) and p62 on s-IBM and control muscle biopsies. Based on conventional stains 36 patients with characteristic clinical features of s-IBM were subclassified as presumed definite s-IBM (d s-IBM, n = 17) or possible s-IBM (p s-IBM, n = 19) according to the presence or absence of vacuolated muscle fibers. Immunohistochemically, TDP-43 and p62 were the most sensitive markers, accumulating in all d s-IBM and in 31% and 37%, respectively, of the p s-IBM cases and thus enabling reclassification of these cases as d s-IBM. We recommend using TDP-43 and p62antibodies in the histological diagnosis workup of s-IBM. The specificity of these markers has to be further validated in prospective series.
- PMID:
- 22106712
- [PubMed - in process]
Cellular prion protein is essential for oligomeric amyloid-β-induced neuronal cell death.
Source
Department of Pathology.
Abstract
In Alzheimer disease (AD), amyloid-β (Aβ) oligomer is suggested to play a critical role in imitating neurodegeneration, although its pathogenic mechanism remains to be determined. Recently, the cellular prion protein (PrP(C)) has been reported to be an essential co-factor in mediating the neurotoxic effect of Aβ oligomer. However, these previous studies focused on the synaptic plasticity in either the presence or the absence of PrP(C) and no study to date has reported whether PrP(C) is required for the neuronal cell death, the most critical element of neurodegeneration in AD. Here, we show that Prnp(-/-) mice are resistant to the neurotoxic effect of Aβ oligomer in vivo and in vitro. Furthermore, application of an anti-PrP(C) antibody or PrP(C) peptide prevents Aβ oligomer-induced neurotoxicity. These findings are the first to demonstrate that PrP(C) is required for Aβ oligomer-induced neuronal cell death, the pathology essential to cognitive loss.
Defects in the medial entorhinal cortex and dentate gyrus in the mouse model of Sanfilippo syndrome type B.
Source
Department of Biological Chemistry, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California, United States of America.
Abstract
Sanfilippo syndrome type B (MPS IIIB) is characterized by profound mental retardation in childhood, dementia and death in late adolescence; it is caused by deficiency of α-N-acetylglucosaminidase and resulting lysosomal storage of heparan sulfate. A mouse model, generated by homologous recombination of the Naglu gene, was used to study pathological changes in the brain. We found earlier that neurons in the medial entorhinal cortex (MEC) and the dentate gyrus showed a number of secondary defects, including the presence of hyperphosphorylated tau (Ptau) detected withantibodies raised against Ptau in Alzheimer disease brain. By further use of immunohistochemistry, we now show staining in neurons of the same area for beta amyloid, extending the resemblance to Alzheimer disease. Ptau inclusions in the dentate gyrus of MPS IIIB mice were reduced in number when the mice were administered LiCl, a specific inhibitor of Gsk3β. Additional proteins found elevated in MEC include proteins involved in autophagy and the heparan sulfate proteoglycans, glypicans 1 and 5, the latter closely related to the primary defect. The level of secondary accumulations was associated with elevation of glypican, as seen by comparing brains of mice at different ages or with different mucopolysaccharide storage diseases. The MEC of an MPS IIIA mouse had the same intense immunostaining for glypican 1 and other markers as MPS IIIB, while MEC of MPS I and MPS II mice had weak staining, and MEC of an MPS VI mouse had no staining at all for the same proteins. A considerable amount of glypican was found in MEC of MPS IIIB mice outside of lysosomes. We propose that it is the extralysosomal glypican that would be harmful to neurons, because its heparan sulfate branches could potentiate the formation of Ptau and beta amyloid aggregates, which would be toxic as well as difficult to degrade.
Quantitation of amyloid beta peptides in CSF by surface enhanced MALDI-TOF.
Source
Wyeth Research, Collegeville, PA, USA. Takahashi.eddie@gmail.com
Abstract
Alzheimer's disease is characterized by the deposition of amyloid plaques in the brain. The major components of these plaques are β-amyloid (Aβ) peptides. The CSF concentration of these peptides can therefore provide a valuable biomarker for potentially predicting the state of disease and/or monitoring the efficacy of a drug aiming to inhibit the formation of amyloid plaques. Although the concentration of a given peptide in CSF can easily be measured by ELISA methods, few methods are able to simultaneously observe and distinguish between various peptides of similar yet slightly different amino acid composition. The Surface Enhanced Laser Desorption/Ionization-Time Of Flight mass spectrometry (SELDI-TOF) technology, a platform combining the use of an antibody and MALDI-TOF, can be used to simultaneously detect and quantitate various Aβ peptides with sensitivities in the picomolar range.
Aβ inhibition of ionic conductance in mouse basal forebrain neurons is dependent upon the cellular prion protein PrPC.
Source
Department of Medicine-Neurology, University of Alberta, Edmonton, Alberta T6G2S2, Canada.
Abstract
Current therapies for Alzheimer's disease (AD) address a loss of cholinergic neurons, while accumulation of neurotoxicamyloid β (Aβ) peptide assemblies is thought central to molecular pathogenesis. Overlaps may exist between prionopathies and AD wherein Aβ oligomers bind to the cellular prion protein PrP(C) and inhibit synaptic plasticity in the hippocampus (Laurén et al., 2009). Here we applied oligomeric Aβ to neurons with different PrP (Prnp) gene dosage. Whole-cell recordings were obtained from dissociated neurons of the diagonal band of Broca (DBB), a cholinergic basal forebrain nucleus. In wild-type (wt) mice, Aβ₁₋₄₂ evoked a concentration-dependent reduction of whole-cell outward currents in a voltage range between -30 and +30 mV; reduction occurred through a combined modulation of a suite of potassium conductances including the delayed rectifier (I(K)), the transient outward (I(A)), and the iberiotoxin-sensitive (calcium-activated potassium, I(C)) currents. Inhibition was not seen with Aβ₄₂₋₁ peptide, while Aβ₁₋₄₂-induced responses were reduced by application of anti-PrP antibody, attenuated in cells from Prnp⁰/⁺ hemizygotes, and absent in Prnp⁰/⁰ homozygotes. Similarly, amyloidogenic amylin peptide depressed DBB whole-cell currents in DBB cells from wt mice, but not Prnp⁰/⁰ homozygotes. While prior studies give broad support for a neuroprotective function for PrP(C), our data define a latent pro-pathogenic role in the presence of amyloid assemblies.
Detection of Peri-Synaptic Amyloid-β Pyroglutamate Aggregates in Early Stages of Alzheimer's Disease and in AβPP Transgenic Mice Using a Novel Monoclonal Antibody.
Source
AFFiRiS AG, Vienna Biocenter, Vienna, Austria.
Abstract
The neurodegenerative pathology in patients with Alzheimer's disease (AD) has been associated with the progressive accumulation of aggregated and post-translationally modified amyloid-β (Aβ) species. Among them, recent studies indicate that the pyroglutamate modification of Aβ (pE(3)Aβ) catalyzed by glutaminyl cyclase might play an important role in the pathogenesis of AD. Although the effects of the pyroglutamate modification on Aβ aggregation and toxicity have been investigated, less is known about the distribution of pE(3)Aβ across the spectrum of AD and in the brains ofamyloid-β protein precursor (AβPP) transgenic (tg) animals. For this purpose, we generated a novel monoclonal antibody(denominated D129) that specifically recognizes A pE(3)Aβ and characterized the patterns of distribution in the postmortem brains samples from AD patients divided by disease stage (Braak stage) and in AβPP tg mice. We found that in early stages of AD and young AβPP tg mice pE(3)Aβ was found in discrete linear and granular aggregates in the neuropil that co-localized with the pre-synaptic protein synaptophysin and was in close opposition to dendrites labeled with MAP2. In later stages of AD and in older AβPP tg mice, pE(3)Aβ was abundant in diffuse and mature plaques. In conclusion, this study suggests that peri-synaptic accumulation of pE(3)Aβ might contribute to early cognitive dysfunction in AD.
Induction of Methionine-Sulfoxide Reductases Protects Neurons from Amyloidβ-Protein Insults in Vitro and in Vivo.
Source
Department of Pharmacology and Toxicology, School of Pharmacy, University of Kansas , 1251 Wescoe Hall Drive, Lawrence, Kansas 66045, United States.
Abstract
Self-assembly of amyloid β-protein (Aβ) into toxic oligomers and fibrillar polymers is believed to cause Alzheimer's disease (AD). In the AD brain, a high percentage of Aβ contains Met-sulfoxide at position 35, though the role this modification plays in AD is not clear. Oxidation of Met(35) to sulfoxide has been reported to decrease the extent of Aβ assembly and neurotoxicity, whereas surprisingly, oxidation of Met(35) to sulfone yields a toxicity similar to that of unoxidized Aβ. We hypothesized that the lower toxicity of Aβ-sulfoxide might result not only from structural alteration of the C-terminal region but also from activation of methionine-sulfoxide reductase (Msr), an important component of the cellular antioxidant system. Supporting this hypothesis, we found that the low toxicity of Aβ-sulfoxide correlated with induction of Msr activity. In agreement with these observations, in MsrA(-/-) mice the difference in toxicity between native Aβ and Aβ-sulfoxide was essentially eliminated. Subsequently, we found that treatment with N-acetyl-Met-sulfoxide could induce Msr activity and protect neuronal cells from Aβ toxicity. In addition, we measured Msr activity in a double-transgenic mouse model of AD and found that it was increased significantly relative to that of nontransgenic mice. Immunization with a novel Met-sulfoxide-rich antigen for 6 months led to antibody production, decreased Msr activity, and lowered hippocampal plaque burden. The data suggest an important neuroprotective role for the Msr system in the AD brain, which may lead to development of new therapeutic approaches for AD.
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