- Email: [email protected]
PS1 mice
Poster Presentations: P2
of CCF-STTG1 cells. Moreover, recombinant KLK7 protein was capable to degrade Ab. Intriguingly, the conditioned media from rat primary astrocytes, but not neurons, showed the Ab-degrading activity as well as KLK7 expression. This activity was inhibited by anti-KLK7 neutralizing antibody. We are currently investigating effect of gene ablation of KLK7 on brain Ab levels. Conclusions: We identified KLK7 as a novel Ab-degrading protease secreted from astrocytic cells.
P2-058
PHYSIOLOGICALLY RELEVANT NON-TOXIC AMYLOID-LIKE OLIGOMERS OF BRAIN PROTEINS BASP1 AND GAP-43
Oksana S. Forsova1, Vladislav V. Zakharov1,2, Faina M. Zakharova3, Petersburg Nuclear Physics Institute, Gatchina, Russia; 2Saint Petersburg State Polytechnical University, Saint Petersburg, Russia; 3Saint Petersburg State University, Saint Petersburg, Russia. Contact e-mail: vit-oksana@ yandex.ru
1
Background: Amyloid oligomers are currently considered as primary toxic species in Alzheimer’s and other neurodegenerative diseases. On the other hand, amyloid-like aggregates (including oligomers) of some neuronal proteins have been shown to participate in synaptic plasticity and hence to be non-pathological. Previously, we have shown in vitro that presynaptic proteins BASP1 and GAP-43, not associated with any known amyloid disease, form oligomers similar to oligomers of amyloid proteins. In this study, we examined amyloid-like properties and toxicity of BASP1 and GAP-43 oligomers and investigated whether BASP1 and GAP-43 oligomers exist as native membrane-bound complexes. Methods: BASP1 and GAP-43 proteins were purified from bovine brain. Amyloidlike aggregation properties were assessed by electron microscopy, electrophoresis, Congo red and ThT binding assays, dotblotting using the conformation-specific antibodies against amyloid oligomers (A11). Cell toxicity was measured by MTT and WST-8 microplate assays using PC12 cells. Lipid rafts and intact synaptosomes were isolated from rat brain and treated with glutaraldehyde and DTSP, respectively. In situ cross-linked proteins were analyzed by SDS-PAGE in 2.5-20% gradient gels and western blotting. Results: BASP1 and GAP-43 oligomers resemble amyloid protein oligomers by the following features: variation of aggregation number (on average, 10-14 for BASP1 and 5-7 for GAP-43), ion channel activity in planar lipid bilayers, fibril formation, recognition by A11 anti-oligomer antibodies. Nterminal peptide myr-BASP1(1-13) readily forms amyloid-like fibrils. BASP1 and GAP-43 proved to be non-toxic to PC12 cells when applied either as monomers or oligomers, while myrBASP1(1-13) decreased MTT reduction by PC12 cells in a manner similar to amyloid-beta peptides. Cross-linking of rat brain synaptic membranes revealed high molecular weight complexes recognized by antibodies to BASP1 and GAP-43. Nearest neighbor cross-linking analysis revealed that they represent BASP1 and GAP-43 homo- and heterooligomers, also containing calmodulin. The ability of the proteins to form heterooligomers of various stoichiometric composition was confirmed in vitro. Conclusions: BASP1 and GAP-43 exist as lipid raft-associated homo- and heterooligomers. These oligomers may therefore be a physiologically relevant protein form with no cell toxicity, despite its similarity to amyloid oligomers. The results are of great interest for the study of possible functions of diseaserelated amyloid oligomers.
P2-059
P505
ROLE OF AMYLOID PRECURSOR PROTEIN INTERACTING PROTEIN FE65 IN NEURITE OUTGROWTH
Kwok-Fai Lau, Hei Nga Cheung, Ho Yin Edwin Chan, The Chinese University of Hong Kong, Hong Kong, Hong Kong. Contact e-mail: kflau@ cuhk.edu.hk Background: FE65 is a brain-enriched multi-domain adaptor protein which has been shown to interact with the cytoplasmic domain of amyloid precursor protein (APP) and to alter APP metabolism. Additionally, emerging evidence reveals that FE65 is also involved in other neurophysiological processes. As an adaptor protein, FE65 functions in recruiting various interactors to form biological complexes. Hence, identification of the full complement of FE65 interacting proteins is an important step for improving our understanding of FE65. Methods: In this project, we used FE65 PTB1 domain as bait to perform yeast two-hybrid screen. Protein-protein interaction was further analyzed by various biochemical assays. The effect of the novel FE65 interacting protein on neurite outgrowth was analyzed. Results: We showed that via its first phosphotyrosine binding (PTB) domain, FE65 binds to the small GTPase ADP-ribosylation factor 6 (ARF6). Interestingly, FE65 stimulates the activation of both ARF6 and its downstream GTPase Rac1, a regulator of actin dynamics and functions in growth cones to stimulate neurite outgrowth. Furthermore, overexpression of FE65 and/or ARF6 promotes whereas siRNA knockdown of FE65 or ARF6 inhibits neurite outgrowth in cultured neurons as compared to the mock transfected control cells. Moreover, knockdown of ARF6 attenuates FE65 stimulation of neurite outgrowth and defective neurite outgrowth seen in FE65-deficient neurons is partially corrected by ARF6 overexpression. Importantly, the stimulatory effect of FE65 and ARF6 on neurite outgrowth is abrogated either by dominant-negative Rac1 or knockdown of Rac1. Conclusions: We show that FE65 as a novel regulator of neurite outgrowth via controlling ARF6-Rac1 signaling. Severe neurite degeneration is observed in Alzheimer’s disease (AD) which is suggested to be associated with cognitive decline in the disease. Thus, strategies that attenuate neurite loss and/or stimulate neurite outgrowth may represent a promising direction for AD therapeutic development. Our work shed important light on the role of FE65 in neurite outgrowth.
P2-060
THE EFFECT OF EXTRASYNAPTIC NMDAR-2B WITH DAPK1 IMPAIRED THE HIPPOCAMPAL CIRCUIT IN APP/PS1 MICE
Qu Na, Jr.1, Teng Zhang2, Qi Zhang, Sr.2,3, Qing Tian2, 1Wuhan Mental Centre, Huazhong University of Science and Technology, Wuhan, China; 2 Huazhong University of Science and Technology, Wuhan, China; 3Liyuan Hospital, Wuhan, China. Contact e-mail: [email protected] Background: Hippocampus is the key brain region in sentiment
and cognition and shows obvious neural circuit impairment even in the early stage of neurodegenerative disease related, e.g. Alzheimer’s disease (AD). Previous studies indicates the important role of extra-synaptic NMDA receptor 2B (NMDAR-2B) in the hippocampal circuit impairment of AD, and the death-associated kinase protein 1(DAPK1) is the key active factor of extra-synaptic NMDAR-2B. To specific inhibit the activating of extra-synaptic NMDAR-2B without any effect on its physiological function, we designed a peptide (Tat-D-NR2Bi) with fine permeation through the blood-brain barrier and cell membrane which could
P506
Poster Presentations: P2
competitvely binding DAPK1 and decrease the activation of NMDAR-2B by DAPK1. Methods: In this research we will deploy DAPK1-/- mice and APP/PS1 mice, cultured brain slice and cell/ primary neuron to study the role and the mechanism of DAPK1 and NMDAR-2B of DG neuron in the impairment of hippocampal neural circuit with electron-microscope, immunofluorence, electrophysiologic and photo-hereditism techniques. Results: We found that DAPK1 was decreased phosphorlation at Y491/492 in hippocampus and extra-synaptic NMDAR-2B with DAPK1 induced neuron and synapse loss on DG-CA3-CA1 circuit in APP/PS1 mice. However, Tat-D-NR2Bi specific inhibited impairment of extra-synaptic NMDAR-2B with DAPK1 in DG region. Conclusions: The results will help to the understanding the rules of neural circuit impairment under pathological conditions.
subjects showed a significantly higher mean level of phosphorylated tau in comparison to low-risk subjects (36.7061.28 vs 29.1761.58 pg/mL; p < 0.0001). A marked difference in PET imaging measures of amyloid beta load by Standardized Uptake Value Ratios (SUVR) was observed between high- risk subjects and lowrisk subjects. High-risk subjects (n¼40) showed a significantly higher (worse) SUVR measurement of amyloid burden compared to 30 low-risk subjects (mean SUVR 1.9060.06 vs 1.5260.07; p < 0.0001). SUVR shows a strong association with TOMM40 ’523 genotype. On the Mini Mental State Examination, high-risk subjects had a significantly lower mean score than low-risk subjects (26.4560.13 vs 27.3760.16; p ¼ 0.0025). Conclusions: These results show that the high and low BRAA risk categories are associated with statistically significant differences in neurocognitive tests, CSF, and imaging biomarkers between patients diagnosed with MCI or AD and healthy controls. BRAA assignment of risk is consistent with other currently accepted measures of neurocognitive decline and is accurate and less expensive to apply to current drug development, compared to non-validated CSF assays or expensive imaging studies.
P2-062
ARE CEREBROSPINAL FLUID LEVELS OF NEUROGRANIN ASSOCIATED WITH HIPPOCAMPAL ATROPHY AND BRAIN WHITE MATTER VOLUME DECREASE?
Simone Lista1, Enrica Cavedo1,2,3, Henrik Zetterberg4,5, Bruno Dubois1,6, Stephane Epelbaum6, Foudil Lamari7, Kaj Blennow4, Harald Hampel1, 1 Sorbonne Universites, Universite Pierre et Marie Curie-Paris 6, Paris, France; 2CATI Multicenter Neuroimaging Platform, Paris, France; 3IRCCS Istituto Centro San Giovanni di Dio-Fatebenefratelli, Brescia, Italy; 4 Clinical Neurochemistry Laboratory, Institute of Neuroscience and Physiology, the Sahlgrenska Academy at the University of Gothenburg, M€olndal, Sweden; 5UCL Institute of Neurology, London, United Kingdom; 6 APHP, Paris, France; 7Laboratoire de Biochimie, AP-HP, H^opital de la Pitie-Salp^etriere, Paris, France. Contact e-mail: [email protected] P2-061
A MULTI-MODAL COMPARISON OF BIOMARKERS STRATIFIED BY A SIMPLE GENETIC RISK PREDICTION ALGORITHM
Michael W. Lutz1, Daniel K. Burns2, Ann M. Saunders1, Scott Sundseth3, Allen D. Roses2,3,4, 1Duke Bryan ADRC, Durham, NC, USA; 2Zinfandel Pharmaceuticals, Chapel Hill, NC, USA; 3Cabernet Pharmaceuticals, Chapel Hill, NC, USA; 4Duke University School of Medicine, Durham, NC, USA. Contact e-mail: [email protected] Background: A biomarker risk assignment algorithm (BRAA) based on APOE and TOMM40 523 genotypes and age is being used to enrich an international phase 3, double-blind, randomized, placebo-controlled clinical trial. The data from this prospective trial will be used to support qualification of the BRAA by regulatory agencies. This presentation reports a multi-modal comparison of genetic risk estimated by the algorithm and by neurocognitive measures, CSF biomarker levels, and PET imaging measures of amyloid beta load in the Alzheimer’s Disease Neuroimaging Initiative (ADNI) cohort. Methods: Data from ADNI (n ¼ 660) were used with the BRAA to estimate risk of developing MCI due to AD. Risk prediction was compared to multi-modal data including neurocognitive tests, CSF biomarkers (AB and tau), and PET imaging measurement of amyloid burden. Results: For the ADNI cohort, subjects predicted to be high risk by the BRAA showed a significantly lower mean level of CSF AB42 compared with low-risk subjects (157.1563.78 vs 195.5964.67 pg/mL; p < 0.0001). High-risk
Background: Synaptic dysfunction, an early event in the pathogenesis of Alzheimer’s disease (AD), is associated with cognitive decline. Neurogranin (Ng) is a postsynaptic protein implicated in the modulation of calmodulin levels in response to increased intracellular calcium after neuronal excitation. Ng is supposed to be associated with cognitive function. Ng expression is particularly elevated in the associative cortical areas of the normal human brain while lower concentrations are present in the hippocampus and frontal cortex in AD patients. Interestingly, the cerebrospinal fluid (CSF) contains a set of synaptic proteins, including Ng. A pilot study has demonstrated an increase in CSF Ng levels in AD patients versus healthy subjects, thus highlighting the potential role of Ng as an AD biomarker. Furthermore, a significant increase in CSF Ng levels in AD dementia is documented in three independent clinical cohorts and the presence of elevated CSF Ng concentrations – at the Mild Cognitive Impairment stage – is shown to predict progression to dementia due to AD. The aims of this study are: (1) the detection of CSF Ng levels in AD dementia and prodromal AD patients and (2) the association of CSF Ng with hippocampal volume (HV) and brain white matter (WM) volume. Methods: 12 AD dementia and 12 prodromal AD patients have been enrolled in a cross-sectional study. A recently in-house developed ELISA will be utilized to detect CSF Ng levels. A voxel-based morphometry (VBM) analysis using the Statistical Parametric Mapping (SPM8) software will be performed to measure HV and brain WM volume. Results: At the time of the writing the
of CCF-STTG1 cells. Moreover, recombinant KLK7 protein was capable to degrade Ab. Intriguingly, the conditioned media from rat primary astrocytes, but not neurons, showed the Ab-degrading activity as well as KLK7 expression. This activity was inhibited by anti-KLK7 neutralizing antibody. We are currently investigating effect of gene ablation of KLK7 on brain Ab levels. Conclusions: We identified KLK7 as a novel Ab-degrading protease secreted from astrocytic cells.
P2-058
PHYSIOLOGICALLY RELEVANT NON-TOXIC AMYLOID-LIKE OLIGOMERS OF BRAIN PROTEINS BASP1 AND GAP-43
Oksana S. Forsova1, Vladislav V. Zakharov1,2, Faina M. Zakharova3, Petersburg Nuclear Physics Institute, Gatchina, Russia; 2Saint Petersburg State Polytechnical University, Saint Petersburg, Russia; 3Saint Petersburg State University, Saint Petersburg, Russia. Contact e-mail: vit-oksana@ yandex.ru
1
Background: Amyloid oligomers are currently considered as primary toxic species in Alzheimer’s and other neurodegenerative diseases. On the other hand, amyloid-like aggregates (including oligomers) of some neuronal proteins have been shown to participate in synaptic plasticity and hence to be non-pathological. Previously, we have shown in vitro that presynaptic proteins BASP1 and GAP-43, not associated with any known amyloid disease, form oligomers similar to oligomers of amyloid proteins. In this study, we examined amyloid-like properties and toxicity of BASP1 and GAP-43 oligomers and investigated whether BASP1 and GAP-43 oligomers exist as native membrane-bound complexes. Methods: BASP1 and GAP-43 proteins were purified from bovine brain. Amyloidlike aggregation properties were assessed by electron microscopy, electrophoresis, Congo red and ThT binding assays, dotblotting using the conformation-specific antibodies against amyloid oligomers (A11). Cell toxicity was measured by MTT and WST-8 microplate assays using PC12 cells. Lipid rafts and intact synaptosomes were isolated from rat brain and treated with glutaraldehyde and DTSP, respectively. In situ cross-linked proteins were analyzed by SDS-PAGE in 2.5-20% gradient gels and western blotting. Results: BASP1 and GAP-43 oligomers resemble amyloid protein oligomers by the following features: variation of aggregation number (on average, 10-14 for BASP1 and 5-7 for GAP-43), ion channel activity in planar lipid bilayers, fibril formation, recognition by A11 anti-oligomer antibodies. Nterminal peptide myr-BASP1(1-13) readily forms amyloid-like fibrils. BASP1 and GAP-43 proved to be non-toxic to PC12 cells when applied either as monomers or oligomers, while myrBASP1(1-13) decreased MTT reduction by PC12 cells in a manner similar to amyloid-beta peptides. Cross-linking of rat brain synaptic membranes revealed high molecular weight complexes recognized by antibodies to BASP1 and GAP-43. Nearest neighbor cross-linking analysis revealed that they represent BASP1 and GAP-43 homo- and heterooligomers, also containing calmodulin. The ability of the proteins to form heterooligomers of various stoichiometric composition was confirmed in vitro. Conclusions: BASP1 and GAP-43 exist as lipid raft-associated homo- and heterooligomers. These oligomers may therefore be a physiologically relevant protein form with no cell toxicity, despite its similarity to amyloid oligomers. The results are of great interest for the study of possible functions of diseaserelated amyloid oligomers.
P2-059
P505
ROLE OF AMYLOID PRECURSOR PROTEIN INTERACTING PROTEIN FE65 IN NEURITE OUTGROWTH
Kwok-Fai Lau, Hei Nga Cheung, Ho Yin Edwin Chan, The Chinese University of Hong Kong, Hong Kong, Hong Kong. Contact e-mail: kflau@ cuhk.edu.hk Background: FE65 is a brain-enriched multi-domain adaptor protein which has been shown to interact with the cytoplasmic domain of amyloid precursor protein (APP) and to alter APP metabolism. Additionally, emerging evidence reveals that FE65 is also involved in other neurophysiological processes. As an adaptor protein, FE65 functions in recruiting various interactors to form biological complexes. Hence, identification of the full complement of FE65 interacting proteins is an important step for improving our understanding of FE65. Methods: In this project, we used FE65 PTB1 domain as bait to perform yeast two-hybrid screen. Protein-protein interaction was further analyzed by various biochemical assays. The effect of the novel FE65 interacting protein on neurite outgrowth was analyzed. Results: We showed that via its first phosphotyrosine binding (PTB) domain, FE65 binds to the small GTPase ADP-ribosylation factor 6 (ARF6). Interestingly, FE65 stimulates the activation of both ARF6 and its downstream GTPase Rac1, a regulator of actin dynamics and functions in growth cones to stimulate neurite outgrowth. Furthermore, overexpression of FE65 and/or ARF6 promotes whereas siRNA knockdown of FE65 or ARF6 inhibits neurite outgrowth in cultured neurons as compared to the mock transfected control cells. Moreover, knockdown of ARF6 attenuates FE65 stimulation of neurite outgrowth and defective neurite outgrowth seen in FE65-deficient neurons is partially corrected by ARF6 overexpression. Importantly, the stimulatory effect of FE65 and ARF6 on neurite outgrowth is abrogated either by dominant-negative Rac1 or knockdown of Rac1. Conclusions: We show that FE65 as a novel regulator of neurite outgrowth via controlling ARF6-Rac1 signaling. Severe neurite degeneration is observed in Alzheimer’s disease (AD) which is suggested to be associated with cognitive decline in the disease. Thus, strategies that attenuate neurite loss and/or stimulate neurite outgrowth may represent a promising direction for AD therapeutic development. Our work shed important light on the role of FE65 in neurite outgrowth.
P2-060
THE EFFECT OF EXTRASYNAPTIC NMDAR-2B WITH DAPK1 IMPAIRED THE HIPPOCAMPAL CIRCUIT IN APP/PS1 MICE
Qu Na, Jr.1, Teng Zhang2, Qi Zhang, Sr.2,3, Qing Tian2, 1Wuhan Mental Centre, Huazhong University of Science and Technology, Wuhan, China; 2 Huazhong University of Science and Technology, Wuhan, China; 3Liyuan Hospital, Wuhan, China. Contact e-mail: [email protected] Background: Hippocampus is the key brain region in sentiment
and cognition and shows obvious neural circuit impairment even in the early stage of neurodegenerative disease related, e.g. Alzheimer’s disease (AD). Previous studies indicates the important role of extra-synaptic NMDA receptor 2B (NMDAR-2B) in the hippocampal circuit impairment of AD, and the death-associated kinase protein 1(DAPK1) is the key active factor of extra-synaptic NMDAR-2B. To specific inhibit the activating of extra-synaptic NMDAR-2B without any effect on its physiological function, we designed a peptide (Tat-D-NR2Bi) with fine permeation through the blood-brain barrier and cell membrane which could
P506
Poster Presentations: P2
competitvely binding DAPK1 and decrease the activation of NMDAR-2B by DAPK1. Methods: In this research we will deploy DAPK1-/- mice and APP/PS1 mice, cultured brain slice and cell/ primary neuron to study the role and the mechanism of DAPK1 and NMDAR-2B of DG neuron in the impairment of hippocampal neural circuit with electron-microscope, immunofluorence, electrophysiologic and photo-hereditism techniques. Results: We found that DAPK1 was decreased phosphorlation at Y491/492 in hippocampus and extra-synaptic NMDAR-2B with DAPK1 induced neuron and synapse loss on DG-CA3-CA1 circuit in APP/PS1 mice. However, Tat-D-NR2Bi specific inhibited impairment of extra-synaptic NMDAR-2B with DAPK1 in DG region. Conclusions: The results will help to the understanding the rules of neural circuit impairment under pathological conditions.
subjects showed a significantly higher mean level of phosphorylated tau in comparison to low-risk subjects (36.7061.28 vs 29.1761.58 pg/mL; p < 0.0001). A marked difference in PET imaging measures of amyloid beta load by Standardized Uptake Value Ratios (SUVR) was observed between high- risk subjects and lowrisk subjects. High-risk subjects (n¼40) showed a significantly higher (worse) SUVR measurement of amyloid burden compared to 30 low-risk subjects (mean SUVR 1.9060.06 vs 1.5260.07; p < 0.0001). SUVR shows a strong association with TOMM40 ’523 genotype. On the Mini Mental State Examination, high-risk subjects had a significantly lower mean score than low-risk subjects (26.4560.13 vs 27.3760.16; p ¼ 0.0025). Conclusions: These results show that the high and low BRAA risk categories are associated with statistically significant differences in neurocognitive tests, CSF, and imaging biomarkers between patients diagnosed with MCI or AD and healthy controls. BRAA assignment of risk is consistent with other currently accepted measures of neurocognitive decline and is accurate and less expensive to apply to current drug development, compared to non-validated CSF assays or expensive imaging studies.
P2-062
ARE CEREBROSPINAL FLUID LEVELS OF NEUROGRANIN ASSOCIATED WITH HIPPOCAMPAL ATROPHY AND BRAIN WHITE MATTER VOLUME DECREASE?
Simone Lista1, Enrica Cavedo1,2,3, Henrik Zetterberg4,5, Bruno Dubois1,6, Stephane Epelbaum6, Foudil Lamari7, Kaj Blennow4, Harald Hampel1, 1 Sorbonne Universites, Universite Pierre et Marie Curie-Paris 6, Paris, France; 2CATI Multicenter Neuroimaging Platform, Paris, France; 3IRCCS Istituto Centro San Giovanni di Dio-Fatebenefratelli, Brescia, Italy; 4 Clinical Neurochemistry Laboratory, Institute of Neuroscience and Physiology, the Sahlgrenska Academy at the University of Gothenburg, M€olndal, Sweden; 5UCL Institute of Neurology, London, United Kingdom; 6 APHP, Paris, France; 7Laboratoire de Biochimie, AP-HP, H^opital de la Pitie-Salp^etriere, Paris, France. Contact e-mail: [email protected] P2-061
A MULTI-MODAL COMPARISON OF BIOMARKERS STRATIFIED BY A SIMPLE GENETIC RISK PREDICTION ALGORITHM
Michael W. Lutz1, Daniel K. Burns2, Ann M. Saunders1, Scott Sundseth3, Allen D. Roses2,3,4, 1Duke Bryan ADRC, Durham, NC, USA; 2Zinfandel Pharmaceuticals, Chapel Hill, NC, USA; 3Cabernet Pharmaceuticals, Chapel Hill, NC, USA; 4Duke University School of Medicine, Durham, NC, USA. Contact e-mail: [email protected] Background: A biomarker risk assignment algorithm (BRAA) based on APOE and TOMM40 523 genotypes and age is being used to enrich an international phase 3, double-blind, randomized, placebo-controlled clinical trial. The data from this prospective trial will be used to support qualification of the BRAA by regulatory agencies. This presentation reports a multi-modal comparison of genetic risk estimated by the algorithm and by neurocognitive measures, CSF biomarker levels, and PET imaging measures of amyloid beta load in the Alzheimer’s Disease Neuroimaging Initiative (ADNI) cohort. Methods: Data from ADNI (n ¼ 660) were used with the BRAA to estimate risk of developing MCI due to AD. Risk prediction was compared to multi-modal data including neurocognitive tests, CSF biomarkers (AB and tau), and PET imaging measurement of amyloid burden. Results: For the ADNI cohort, subjects predicted to be high risk by the BRAA showed a significantly lower mean level of CSF AB42 compared with low-risk subjects (157.1563.78 vs 195.5964.67 pg/mL; p < 0.0001). High-risk
Background: Synaptic dysfunction, an early event in the pathogenesis of Alzheimer’s disease (AD), is associated with cognitive decline. Neurogranin (Ng) is a postsynaptic protein implicated in the modulation of calmodulin levels in response to increased intracellular calcium after neuronal excitation. Ng is supposed to be associated with cognitive function. Ng expression is particularly elevated in the associative cortical areas of the normal human brain while lower concentrations are present in the hippocampus and frontal cortex in AD patients. Interestingly, the cerebrospinal fluid (CSF) contains a set of synaptic proteins, including Ng. A pilot study has demonstrated an increase in CSF Ng levels in AD patients versus healthy subjects, thus highlighting the potential role of Ng as an AD biomarker. Furthermore, a significant increase in CSF Ng levels in AD dementia is documented in three independent clinical cohorts and the presence of elevated CSF Ng concentrations – at the Mild Cognitive Impairment stage – is shown to predict progression to dementia due to AD. The aims of this study are: (1) the detection of CSF Ng levels in AD dementia and prodromal AD patients and (2) the association of CSF Ng with hippocampal volume (HV) and brain white matter (WM) volume. Methods: 12 AD dementia and 12 prodromal AD patients have been enrolled in a cross-sectional study. A recently in-house developed ELISA will be utilized to detect CSF Ng levels. A voxel-based morphometry (VBM) analysis using the Statistical Parametric Mapping (SPM8) software will be performed to measure HV and brain WM volume. Results: At the time of the writing the