- Email: [email protected]
Domain Specific Cognitive Patterns (dscp): Identifying Unique Cognitive Profiles in Patients With Dementia and MCI
e50
Hot Topics
by the same person, conducted in a location convenient for participants. Proper compensation was provided. Focus groups were semi-structured, lasting for an hour. Data was recorded by a stenographer; all sessions were tape-recorded and transcribed. Data was analyzed using qualitative computer-based analysis programs designed to extract common themes from the focus groups. Results: 83% of the participants were female; 17% male 48% were between the ages of 55-64; 26% were 65-74; 22% were 75-84; 4% were older than 85. 22% of participants completed less than high school; 30% completed high school; 35% completed some college; 4% have a college degree only; 9% have an advanced degree. 61% have household incomes of less than $20,000 a year; 22% has household income between 20,000- 39,000; 17% 40,000 and above. 13% were employed; 87% were retired.
oligomers of pGlu-Ab3-42 are cytotoxic at an ˜5-fold lower concentration than Ab1-42, and that incubation of pGlu-Ab3-42 with 19-fold molar excess of Ab1-42 yields amyloid cocktails at 100nM total concentration, which is 10-50fold more cytotoxic than either peptide individually. Furthermore, primary neurons from tau knockout mice were insensitive to oligomers made from either pure or pGlu-Ab3-42-containing mixtures. Passive immunization against it in aged APP/PS1dE9 mice significantly reduced general Ab-immunoreactivity. Conclusions: pGlu-Ab stimulates tau-dependent neurotoxicity and inflammation which can be ameliorated by QC-inhibition. Removal of general and fibrillar Ab by anti-pGlu-Ab immunotherapy supports seeding by pGlu-Ab. Based on these data, Probiodrug has launched a drug development program which has entered the regulatory phase.
44% married, 39% widowed, 13% divorced. An analysis of the decision making process highlighted the presence of specific key parameters:
P4-096
#1 Trust of the provider making the request #2 Input of the primary medical doctor (if not the one making the request) #3 Distrust to participate in preliminary drug development #4 Required level of involvement and risk Other factors will also be discussed. Conclusions: Against available anecdotal information, our data suggest the presence of key elements related to the clinical trial participation decision-making process. Addressing such issues in well-designed interventions could be the answer to overcome the lack of diversity in clinical trials that challenges researchers. P4-094
PYROGLUTAMATED b-AMYLOID IS TOXIC, HIGHLY ABUNDANT IN THE ALZHEIMER’S BRAIN, AMPLIFIES TAU-DEPENDENT b-AMYLOID CYTOTOXICITY AND CAN BE ATTENUATED BY PASSIVE IMMUNIZATION OR INHIBITION OF GLUTAMINYL CYCLASE.
Hans-Ulrich Demuth1, George S. Bloom2, Cynthia A. Lemere3, 1Probiodrug AG, Halle(Saale), Germany; 2University of Virginia, Department of Biology, Gilmer Hall, Charlottesville, VA, USA; 3Brigham & Women’s Hospital, Harvard Medical School, Center for Neurologic Diseases NRB 636F, Boston, MA, USA. Contact e-mail: hans-ulrich.demuth@probiodrug. de. Background: Glutaminyl cyclase (QC) facilitates N-pyroglutamate-formation of b-amyloid (pGlu-Ab) promoting neurotoxicity. QC inhibition reduced cognitive impairment, as well as both pGlu-Ab and total plaque load in conventional APP transgenic (tg) mice (Schilling, 2008). Here, we created mice producing primarily toxic pGlu-Ab. In addition, we determined whether pGlu-Ab impacts tau toxicity using primary neurons of wildtype and of tau-KO mice. Lastly, as pGlu-Ab is either deposited in plaques or constantly formed as soluble oligomers, immunization against it or treatment by QC-inhibitors should reduce total plaque load and improve behavior. Methods: pGlu-Ab was expressed in secretory or in lysosomal/endosomal vesicles of novel tg mice. Brains were examined for total and pGluAb load, and behavior screened using a comprehensive testing battery. Immunoreactivity for general Ab and pGlu-Ab was compared in brain of aged humans (including AD), non-human primates, and transgenic AD-like mouse models. Passive immunization with anti-pGlu-Ab mAb’s was performed in aged APP/PS1 mice with AD-like pathology for neuropathological analyses. Results: The new mice display age-dependent neuron loss, gliosis, Ab-deposits and behavioral deficits, all related to the onset of pGlu-Ab-expression. In human brain immunohistochemistry revealed close co-localization of pGlu-Ab and general Ab deposits but less pGlu-Ab relative to general Ab deposits in monkey and AD-like mouse brain. Interestingly, tau-pathology and neuronal loss are also less obvious in these models compared to human AD brain. Using synthetic pGlu-Ab and conventional Ab to treat primary cortical neurons in culture, we report that apparent
TIME COURSE OF COGNITIVE DECLINE IN SUBJECTS WITH MILD ALZHEIMER’S DISEASE BASED ON ADAS-COG SUBSCALES AND NEUROPSYCHOLOGICAL TESTS MEASURED IN ADNI
Suzanne B. Hendrix, Brian M. Wells, Pentara Corporation, Salt Lake City, UT, USA. Contact e-mail: [email protected]. Background: Biomarkers are often used as primary outcomes in proof of concept studies in mild patients rather than ADAS-cog, due to improved sensitivity to change over time. Myriad and Elan measured an alternate cognitive instrument, referred to as an NTB, to improve sensitivity, but it was shown to be comparably sensitive to the ADAS-cog. A cognitive outcome with improved sensitivity would be helpful in a proof of concept study in mild subjects. The ADNI dataset provides useful data for exploring this possibility. Methods: Each separate item on the 13 item ADAS-cog, as well as the neuropsychological test items collected in ADNI were assessed to determine changes at different stages of Alzheimer’s disease. Reverse “S-curves” show the progression on each item relative to different measures of disease progression. Composite scores include items that measure change in different cognitive domains, or at different times within mild disease, and are assessed for sensitivity to progression over time. Results: Four items were identified as changing in the very early stages of AD and quickly reaching a ceiling: Rey Auditory Verbal delayed score, Logical Memory Test - delayed and immediate, and the Delayed Word Recall. Four items were identified as changing steadily across mild and moderate stages of the disease: Digit Span Test, Rey Auditory Verbal Word Recognition, Clock Drawing Test, and Orientation. Three tests showed unusually large variability that was visible in the S-curves: Number Cancellation, Naming Objects and Fingers, and Trail Making Part ATime. A composite score constructed from ADAS-cog and NTB items has substantially improved sensitivity to change in Mild AD, resulting in better power for small sample sizes. Conclusions: Understanding which cognitive items change in mild Alzheimer’s disease enables us to construct composite scores with improved sensitivity to change for use in proof of concept studies. Studies measuring cognitive outcomes could be reanalyzed to assess whether differences were by chance, or are seen on a more robust measure of cognitive change. P4-097
DOMAIN SPECIFIC COGNITIVE PATTERNS (DSCP): IDENTIFYING UNIQUE COGNITIVE PROFILES IN PATIENTS WITH DEMENTIA AND MCI
Rex L. Cannon, John H. Dougherty, Jr, Monica K. Crane, Andrew Dougherty, University of Tennessee Medical Center, Knoxville, TN, USA. Contact e-mail: [email protected]. Background: Recent work by our laboratory resulted in an accurate computer-based tool for detecting early cognitive abnormalities in mild cognitive impairment (MCI) and various stages of Alzheimer’s disease (AD). This Internet accessible screening tool, Computerized Self Test (CST), identified abnormalities in multiple cognitive domains of patients with cognitive
Hot Topics impairment as compared to controls. In recent years, a variety of methods have been proposed to differentially classify patients presenting with memory problems. We sought to develop models that would be efficient in classifying individual patients with significant accuracy and ease of use across laboratories and clinical settings. Methods: We proposed to return to the basic components of human cognition and their invariable correlative structure. We utilized the CST in a sample of 229 (99 male) normal individuals and 140 (59 male) cognitively impaired (CI) individuals. We evaluated fundamental properties of statistical paradigms for classification of patients with cognitive abnormalities. Results: Multivariate statistics provide ample power for detecting and plotting the degree of deficit from the normal population in our study, in essence creating an individualized domain specific cognitive pattern (DSCP). Discriminant analysis provides an effective means for classification of these identified cognitive patterns in individual patients compared to control subjects. Conclusions: Domain Specific Cognitive Patterns may provide an accurate means to differentiate between normal, mild cognitive impairment and stages within AD. Importantly, this type of technology may be utilized in differentiating AD or MCI from other types of dementias, trauma or stroke, thereby improving diagnostic accuracy. P4-098
POTENTIAL OF SAC (S-ALLYL-L-CYSTEINE), A COMPONENT OF GARLIC, AS A NOVEL DRUG TARGET FOR ALZHEIMER’S DISEASE (AD) BASED ON STUDIES FROM PRIMARY NEURONAL CULTURES AND IN VIVO
Balmiki Ray1, Neelima B. Chauhan2, Debomoy K. Lahiri1, Indiana University School of Medicine, Department of Psychiatry, Indianapolis, IN, USA; 2Jesse Brown VA Medical Center Chicago, University of Illinois at Chicago, Chicago, IL, USA. Contact e-mail: rayb@ iupui.edu.
P4-099
e51 PROTECTIVE ROLES OF SNITROSOGLUTATHIONE (GSNO) IN RAT CHRONIC CEREBRAL HYPOPERFUSIONINDUCED MILD COGNITIVE IMPAIRMENT
Singh Inderjit, Medical University of South Carolina, Charleston, SC, USA. Contact e-mail: [email protected]. Background: Early decrease in cerebral blood perfusion featuring in most Alzheimer’s disease (AD) cases has documented as a critical risk for AD. Methods: Because loss of cerebral vascular function is critical for development of AD pathologies under chronic cerebral hypoperfusion (CCH) conditions, we examined if S-nitroso-glutathione (GSNO) is able to improve damaged learning-memory function and Ab accumulation in the brains of aged rats treated with bilateral common carotid artery occlusion (BCCAO), an animal model for human CCH. GSNO is a most abundant low molecular weight S-nitrosothiol and has been regarded as an important endogenous NO source exerting anti-inflammatory and vasoprotective activities. Results: We here report that long-term administration of GSNO (50mg/kg/day for 4 months) improved learning and memory functions of BCCAO treated rats with reducing brain Ab levels. GSNO inhibited proinflammatory signaling in bEnd3 brain endothelial cells and thus reduced gene expression related to endothelial inflammation (i.e. ICAM-1, VCAM, and MMP-9). In addition, GSNO increased uptake of Ab by cultured bEnd3 cell with increasing S-nitrosylation of dynamin-2 protein, a protein regulating cellular endocytosis activity. Conclusions: Taken together, these data first time document a potential therapeutic activity of GSNO on neurovascular pathologies involved in CCH and AD.
1
Background: AD is characterized by amyloid b-peptide (Ab) plaques and neurofibrillary tangles. Ab produces reactive oxygen species (ROS) causing neuroinflammation and neuronal damage. Anticholinesterase drugs are predominantly used for AD but they fail to cure AD. Thus, alternative medications should be developed. From ancient times, garlic (Allium sativum) is used to treat diabetes and cardiovascular disorders. In AD models, we successfully tested a unique preparation, “Aged Garlic Extract” (AGE). AGE contains DAD (Diallyl disulfide) and SAC (S-allyl-L-cysteine). Here we demonstrate novel property of AGE and SAC in neuroprotection, neuropreservation and synaptic integrity. Methods: For neuroprotection, differentiated neuronal PC12 cells were co-treated with 200mM of hydrogen peroxide and either AGE, DAD or SAC. Western immunoblotiing and choline acetyltransferase (ChAT) assays were performed with cell lysates. Cells were also fixed with 4% paraformaldehyde for morphology analyses by ICC. For neurorescue experiments, differentiated neuronal cells were pretreated with AGE, DAD or SAC alone for 48 hours and thereafter, posttreated with ROS alone for 24 hours. Cells were harvested and fixed. Further, APP-Tg2576 mice were fed with special diet containing AGE, SAC or isocaloric control diet for 4 months. Animals were sacrificed and brain was dissected, homogenized and brain homogenates were subjected to Western immunoblotting and ELISA. Results: We observed significant neuroprotective and neurorescue properties of AGE and SAC independently against ROS- mediated insults to neuronal cells. Both AGE and SAC preserved pre-synaptic proteins (SYPH, SNAP-25) in Alzheimer’s APP-Tg mice and ROS-insulted neuronal cells. Notably, AGE increased levels of ChAT activity in ROS-challenge. Conclusions: AGE or SAC treatment protects neurons from ROS-mediated oxidative stress and preserves pre-synaptic proteins. AGE treatment reverses ROS-mediated decline in cholinergic function of neuronal cells by increasing neuronal ChAT activity. Synaptic damage from excessive Ab deposition in APP-Tg mice brain was also prevented by oral AGE or SAC treatment. The pleiotropic effects of AGE and the most active ingredient SAC, can be used as a potential therapeutic agent in treating AD. These findings warrant evaluation of SAC’s clinical potency in larger clinical settings.
P4-100
SMALL MOLECULE INHIBITOR OF AMYLOID BETA-PROTEIN OLIGOMERIZATION REDUCES BRAIN ABETA LOAD IN AN ALZHEIMER’S DISEASE MOUSE MODEL
Aida Attar1, Sharmistha Sinha1, Panchanan Maiti1, Fusheng Yang1, Dana Gant1, Mychica Jones1, Peter Talbiersky2, Thomas Schrader2, Frank-Gerrit Kla¨rner2, Sally Frautschy1, Gal Bitan1, 1UCLA, Los Angeles, CA, USA; 2Universita¨t Duisburg-Essen, Essen, Germany. Contact e-mail: [email protected]. Background: Cognitive dysfunction and brain atrophy seen in Alzheimer’s disease (AD) are believed to be results of a cascade of events beginning with self-assembly of amyloid beta-protein (Abeta) leading to synaptic injury. Formation of Abeta oligomers is mediated by specific electrostatic and hydrophobic interactions that can be targeted to prevent neurotoxicity. Methods: We have identified a small molecule, termed CLR01, that complexes with Abeta and competes for the molecular interactions controlling initial Abeta folding and assembly. We used a variety of in vitro and in vivo assays to test the efficacy of CLR01. Results: Dot blot experiments with the Abeta oligomer specific antibody, A11, show inhibition of oligomer formation and cell culture experiments show significant protection against Abeta-induced toxicity upon administration of CLR01. Initial in vivo experiments have been performed in triple-transgenic and doubletransgenic mouse models of AD. These experiments show significant brain-Abeta load reduction and improvement of spatial working memory following 28 days of subcutaneous CLR01 administration. Conclusions: These data support CLR01 as a promising lead for disease-modifying AD therapy. P4-101
DIAGNOSTIC DISCRIMINATION OF MILD COGNITIVE IMPAIRMENT, ALZHEIMER’S DISEASE, AND OTHER DEMENTIA GROUPS USING THE COGSTATE CLINIC BATTERY
Dustin Hammers, Elizabeth Spurgeon, Kelly Ryan, Carol Persad, Kenya Talton, Tarin Coulas, Arijit Bhaumik, Aviva Nathan, Judith Heidebrink, Nancy Barbas, Bruno Giordani, University of Michigan, Ann Arbor, MI, USA. Contact e-mail: [email protected].
Hot Topics
by the same person, conducted in a location convenient for participants. Proper compensation was provided. Focus groups were semi-structured, lasting for an hour. Data was recorded by a stenographer; all sessions were tape-recorded and transcribed. Data was analyzed using qualitative computer-based analysis programs designed to extract common themes from the focus groups. Results: 83% of the participants were female; 17% male 48% were between the ages of 55-64; 26% were 65-74; 22% were 75-84; 4% were older than 85. 22% of participants completed less than high school; 30% completed high school; 35% completed some college; 4% have a college degree only; 9% have an advanced degree. 61% have household incomes of less than $20,000 a year; 22% has household income between 20,000- 39,000; 17% 40,000 and above. 13% were employed; 87% were retired.
oligomers of pGlu-Ab3-42 are cytotoxic at an ˜5-fold lower concentration than Ab1-42, and that incubation of pGlu-Ab3-42 with 19-fold molar excess of Ab1-42 yields amyloid cocktails at 100nM total concentration, which is 10-50fold more cytotoxic than either peptide individually. Furthermore, primary neurons from tau knockout mice were insensitive to oligomers made from either pure or pGlu-Ab3-42-containing mixtures. Passive immunization against it in aged APP/PS1dE9 mice significantly reduced general Ab-immunoreactivity. Conclusions: pGlu-Ab stimulates tau-dependent neurotoxicity and inflammation which can be ameliorated by QC-inhibition. Removal of general and fibrillar Ab by anti-pGlu-Ab immunotherapy supports seeding by pGlu-Ab. Based on these data, Probiodrug has launched a drug development program which has entered the regulatory phase.
44% married, 39% widowed, 13% divorced. An analysis of the decision making process highlighted the presence of specific key parameters:
P4-096
#1 Trust of the provider making the request #2 Input of the primary medical doctor (if not the one making the request) #3 Distrust to participate in preliminary drug development #4 Required level of involvement and risk Other factors will also be discussed. Conclusions: Against available anecdotal information, our data suggest the presence of key elements related to the clinical trial participation decision-making process. Addressing such issues in well-designed interventions could be the answer to overcome the lack of diversity in clinical trials that challenges researchers. P4-094
PYROGLUTAMATED b-AMYLOID IS TOXIC, HIGHLY ABUNDANT IN THE ALZHEIMER’S BRAIN, AMPLIFIES TAU-DEPENDENT b-AMYLOID CYTOTOXICITY AND CAN BE ATTENUATED BY PASSIVE IMMUNIZATION OR INHIBITION OF GLUTAMINYL CYCLASE.
Hans-Ulrich Demuth1, George S. Bloom2, Cynthia A. Lemere3, 1Probiodrug AG, Halle(Saale), Germany; 2University of Virginia, Department of Biology, Gilmer Hall, Charlottesville, VA, USA; 3Brigham & Women’s Hospital, Harvard Medical School, Center for Neurologic Diseases NRB 636F, Boston, MA, USA. Contact e-mail: hans-ulrich.demuth@probiodrug. de. Background: Glutaminyl cyclase (QC) facilitates N-pyroglutamate-formation of b-amyloid (pGlu-Ab) promoting neurotoxicity. QC inhibition reduced cognitive impairment, as well as both pGlu-Ab and total plaque load in conventional APP transgenic (tg) mice (Schilling, 2008). Here, we created mice producing primarily toxic pGlu-Ab. In addition, we determined whether pGlu-Ab impacts tau toxicity using primary neurons of wildtype and of tau-KO mice. Lastly, as pGlu-Ab is either deposited in plaques or constantly formed as soluble oligomers, immunization against it or treatment by QC-inhibitors should reduce total plaque load and improve behavior. Methods: pGlu-Ab was expressed in secretory or in lysosomal/endosomal vesicles of novel tg mice. Brains were examined for total and pGluAb load, and behavior screened using a comprehensive testing battery. Immunoreactivity for general Ab and pGlu-Ab was compared in brain of aged humans (including AD), non-human primates, and transgenic AD-like mouse models. Passive immunization with anti-pGlu-Ab mAb’s was performed in aged APP/PS1 mice with AD-like pathology for neuropathological analyses. Results: The new mice display age-dependent neuron loss, gliosis, Ab-deposits and behavioral deficits, all related to the onset of pGlu-Ab-expression. In human brain immunohistochemistry revealed close co-localization of pGlu-Ab and general Ab deposits but less pGlu-Ab relative to general Ab deposits in monkey and AD-like mouse brain. Interestingly, tau-pathology and neuronal loss are also less obvious in these models compared to human AD brain. Using synthetic pGlu-Ab and conventional Ab to treat primary cortical neurons in culture, we report that apparent
TIME COURSE OF COGNITIVE DECLINE IN SUBJECTS WITH MILD ALZHEIMER’S DISEASE BASED ON ADAS-COG SUBSCALES AND NEUROPSYCHOLOGICAL TESTS MEASURED IN ADNI
Suzanne B. Hendrix, Brian M. Wells, Pentara Corporation, Salt Lake City, UT, USA. Contact e-mail: [email protected]. Background: Biomarkers are often used as primary outcomes in proof of concept studies in mild patients rather than ADAS-cog, due to improved sensitivity to change over time. Myriad and Elan measured an alternate cognitive instrument, referred to as an NTB, to improve sensitivity, but it was shown to be comparably sensitive to the ADAS-cog. A cognitive outcome with improved sensitivity would be helpful in a proof of concept study in mild subjects. The ADNI dataset provides useful data for exploring this possibility. Methods: Each separate item on the 13 item ADAS-cog, as well as the neuropsychological test items collected in ADNI were assessed to determine changes at different stages of Alzheimer’s disease. Reverse “S-curves” show the progression on each item relative to different measures of disease progression. Composite scores include items that measure change in different cognitive domains, or at different times within mild disease, and are assessed for sensitivity to progression over time. Results: Four items were identified as changing in the very early stages of AD and quickly reaching a ceiling: Rey Auditory Verbal delayed score, Logical Memory Test - delayed and immediate, and the Delayed Word Recall. Four items were identified as changing steadily across mild and moderate stages of the disease: Digit Span Test, Rey Auditory Verbal Word Recognition, Clock Drawing Test, and Orientation. Three tests showed unusually large variability that was visible in the S-curves: Number Cancellation, Naming Objects and Fingers, and Trail Making Part ATime. A composite score constructed from ADAS-cog and NTB items has substantially improved sensitivity to change in Mild AD, resulting in better power for small sample sizes. Conclusions: Understanding which cognitive items change in mild Alzheimer’s disease enables us to construct composite scores with improved sensitivity to change for use in proof of concept studies. Studies measuring cognitive outcomes could be reanalyzed to assess whether differences were by chance, or are seen on a more robust measure of cognitive change. P4-097
DOMAIN SPECIFIC COGNITIVE PATTERNS (DSCP): IDENTIFYING UNIQUE COGNITIVE PROFILES IN PATIENTS WITH DEMENTIA AND MCI
Rex L. Cannon, John H. Dougherty, Jr, Monica K. Crane, Andrew Dougherty, University of Tennessee Medical Center, Knoxville, TN, USA. Contact e-mail: [email protected]. Background: Recent work by our laboratory resulted in an accurate computer-based tool for detecting early cognitive abnormalities in mild cognitive impairment (MCI) and various stages of Alzheimer’s disease (AD). This Internet accessible screening tool, Computerized Self Test (CST), identified abnormalities in multiple cognitive domains of patients with cognitive
Hot Topics impairment as compared to controls. In recent years, a variety of methods have been proposed to differentially classify patients presenting with memory problems. We sought to develop models that would be efficient in classifying individual patients with significant accuracy and ease of use across laboratories and clinical settings. Methods: We proposed to return to the basic components of human cognition and their invariable correlative structure. We utilized the CST in a sample of 229 (99 male) normal individuals and 140 (59 male) cognitively impaired (CI) individuals. We evaluated fundamental properties of statistical paradigms for classification of patients with cognitive abnormalities. Results: Multivariate statistics provide ample power for detecting and plotting the degree of deficit from the normal population in our study, in essence creating an individualized domain specific cognitive pattern (DSCP). Discriminant analysis provides an effective means for classification of these identified cognitive patterns in individual patients compared to control subjects. Conclusions: Domain Specific Cognitive Patterns may provide an accurate means to differentiate between normal, mild cognitive impairment and stages within AD. Importantly, this type of technology may be utilized in differentiating AD or MCI from other types of dementias, trauma or stroke, thereby improving diagnostic accuracy. P4-098
POTENTIAL OF SAC (S-ALLYL-L-CYSTEINE), A COMPONENT OF GARLIC, AS A NOVEL DRUG TARGET FOR ALZHEIMER’S DISEASE (AD) BASED ON STUDIES FROM PRIMARY NEURONAL CULTURES AND IN VIVO
Balmiki Ray1, Neelima B. Chauhan2, Debomoy K. Lahiri1, Indiana University School of Medicine, Department of Psychiatry, Indianapolis, IN, USA; 2Jesse Brown VA Medical Center Chicago, University of Illinois at Chicago, Chicago, IL, USA. Contact e-mail: rayb@ iupui.edu.
P4-099
e51 PROTECTIVE ROLES OF SNITROSOGLUTATHIONE (GSNO) IN RAT CHRONIC CEREBRAL HYPOPERFUSIONINDUCED MILD COGNITIVE IMPAIRMENT
Singh Inderjit, Medical University of South Carolina, Charleston, SC, USA. Contact e-mail: [email protected]. Background: Early decrease in cerebral blood perfusion featuring in most Alzheimer’s disease (AD) cases has documented as a critical risk for AD. Methods: Because loss of cerebral vascular function is critical for development of AD pathologies under chronic cerebral hypoperfusion (CCH) conditions, we examined if S-nitroso-glutathione (GSNO) is able to improve damaged learning-memory function and Ab accumulation in the brains of aged rats treated with bilateral common carotid artery occlusion (BCCAO), an animal model for human CCH. GSNO is a most abundant low molecular weight S-nitrosothiol and has been regarded as an important endogenous NO source exerting anti-inflammatory and vasoprotective activities. Results: We here report that long-term administration of GSNO (50mg/kg/day for 4 months) improved learning and memory functions of BCCAO treated rats with reducing brain Ab levels. GSNO inhibited proinflammatory signaling in bEnd3 brain endothelial cells and thus reduced gene expression related to endothelial inflammation (i.e. ICAM-1, VCAM, and MMP-9). In addition, GSNO increased uptake of Ab by cultured bEnd3 cell with increasing S-nitrosylation of dynamin-2 protein, a protein regulating cellular endocytosis activity. Conclusions: Taken together, these data first time document a potential therapeutic activity of GSNO on neurovascular pathologies involved in CCH and AD.
1
Background: AD is characterized by amyloid b-peptide (Ab) plaques and neurofibrillary tangles. Ab produces reactive oxygen species (ROS) causing neuroinflammation and neuronal damage. Anticholinesterase drugs are predominantly used for AD but they fail to cure AD. Thus, alternative medications should be developed. From ancient times, garlic (Allium sativum) is used to treat diabetes and cardiovascular disorders. In AD models, we successfully tested a unique preparation, “Aged Garlic Extract” (AGE). AGE contains DAD (Diallyl disulfide) and SAC (S-allyl-L-cysteine). Here we demonstrate novel property of AGE and SAC in neuroprotection, neuropreservation and synaptic integrity. Methods: For neuroprotection, differentiated neuronal PC12 cells were co-treated with 200mM of hydrogen peroxide and either AGE, DAD or SAC. Western immunoblotiing and choline acetyltransferase (ChAT) assays were performed with cell lysates. Cells were also fixed with 4% paraformaldehyde for morphology analyses by ICC. For neurorescue experiments, differentiated neuronal cells were pretreated with AGE, DAD or SAC alone for 48 hours and thereafter, posttreated with ROS alone for 24 hours. Cells were harvested and fixed. Further, APP-Tg2576 mice were fed with special diet containing AGE, SAC or isocaloric control diet for 4 months. Animals were sacrificed and brain was dissected, homogenized and brain homogenates were subjected to Western immunoblotting and ELISA. Results: We observed significant neuroprotective and neurorescue properties of AGE and SAC independently against ROS- mediated insults to neuronal cells. Both AGE and SAC preserved pre-synaptic proteins (SYPH, SNAP-25) in Alzheimer’s APP-Tg mice and ROS-insulted neuronal cells. Notably, AGE increased levels of ChAT activity in ROS-challenge. Conclusions: AGE or SAC treatment protects neurons from ROS-mediated oxidative stress and preserves pre-synaptic proteins. AGE treatment reverses ROS-mediated decline in cholinergic function of neuronal cells by increasing neuronal ChAT activity. Synaptic damage from excessive Ab deposition in APP-Tg mice brain was also prevented by oral AGE or SAC treatment. The pleiotropic effects of AGE and the most active ingredient SAC, can be used as a potential therapeutic agent in treating AD. These findings warrant evaluation of SAC’s clinical potency in larger clinical settings.
P4-100
SMALL MOLECULE INHIBITOR OF AMYLOID BETA-PROTEIN OLIGOMERIZATION REDUCES BRAIN ABETA LOAD IN AN ALZHEIMER’S DISEASE MOUSE MODEL
Aida Attar1, Sharmistha Sinha1, Panchanan Maiti1, Fusheng Yang1, Dana Gant1, Mychica Jones1, Peter Talbiersky2, Thomas Schrader2, Frank-Gerrit Kla¨rner2, Sally Frautschy1, Gal Bitan1, 1UCLA, Los Angeles, CA, USA; 2Universita¨t Duisburg-Essen, Essen, Germany. Contact e-mail: [email protected]. Background: Cognitive dysfunction and brain atrophy seen in Alzheimer’s disease (AD) are believed to be results of a cascade of events beginning with self-assembly of amyloid beta-protein (Abeta) leading to synaptic injury. Formation of Abeta oligomers is mediated by specific electrostatic and hydrophobic interactions that can be targeted to prevent neurotoxicity. Methods: We have identified a small molecule, termed CLR01, that complexes with Abeta and competes for the molecular interactions controlling initial Abeta folding and assembly. We used a variety of in vitro and in vivo assays to test the efficacy of CLR01. Results: Dot blot experiments with the Abeta oligomer specific antibody, A11, show inhibition of oligomer formation and cell culture experiments show significant protection against Abeta-induced toxicity upon administration of CLR01. Initial in vivo experiments have been performed in triple-transgenic and doubletransgenic mouse models of AD. These experiments show significant brain-Abeta load reduction and improvement of spatial working memory following 28 days of subcutaneous CLR01 administration. Conclusions: These data support CLR01 as a promising lead for disease-modifying AD therapy. P4-101
DIAGNOSTIC DISCRIMINATION OF MILD COGNITIVE IMPAIRMENT, ALZHEIMER’S DISEASE, AND OTHER DEMENTIA GROUPS USING THE COGSTATE CLINIC BATTERY
Dustin Hammers, Elizabeth Spurgeon, Kelly Ryan, Carol Persad, Kenya Talton, Tarin Coulas, Arijit Bhaumik, Aviva Nathan, Judith Heidebrink, Nancy Barbas, Bruno Giordani, University of Michigan, Ann Arbor, MI, USA. Contact e-mail: [email protected].