- Email: [email protected]
TARGETING THE SHARED PATHOLOGICAL CONFORMERS OF BOTH Aβ AND HYPERPHOSPHYLATED TAU WITH A CONFORMATIONALLY SELECTIVE MONOCLONAL ANTIBODY
Oral Sessions: O3-14: Preclinical Therapeutics III: Tau Plus 4 Ludwig-Maximilians-Universit€at M€unchen, Munich, Germany; 5DZNE, Bonn, Germany. Contact e-mail: [email protected]
Background: Pathological tau aggregation leading to filamentous tau inclusions characterizes neurodegenerative tauopathies such as Alzheimer’s Disease and frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17). Tau aggregation is linked with clinical symptoms and probably represents the mediator of neurodegeneration. At the moment there is no cure for these diseases and existing treatments are only of palliative nature. Methods: Transgenic mice that overexpress human P301S tau mutation resemble many neuropathological features of human tauopathies in combination with behavioral deficits and increased mortality. We tested the effect of the small molecule anle138b as an anti-aggregative drug on the neuropathology and behavioral deficits in P301S mice. The compound was administered from weaning until death. Results: Here we show that reducing the amount of tau aggregates using the compound anle138b ameliorates disease symptoms in a mouse model for tauopathies. Treatment with anle138b significantly prolonged the survival of transgenic mice and rescued behavioral deficits as well as synapse and neuron loss in the CA3 region and stratum lucidum of the hippocampus. Our results indicate that reducing tau pathology with the anti-aggregative compound anle138b represents a clinically effective and promising approach for the treatment of human tauopathies. Conclusions: In conclusion, anle138b mitigate tau pathology and may represent a new lead compound for developing a therapy for human tauopathies. O3-14-02
TARGETING THE SHARED PATHOLOGICAL CONFORMERS OF BOTH Ab AND HYPERPHOSPHYLATED TAU WITH A CONFORMATIONALLY SELECTIVE MONOCLONAL ANTIBODY
Thomas Wisniewski1, Eleanor Drummond2, Krystal Herline2, Yanjie Sun2, Fernando Goni2, 1New York University School of Medicine, New York, New York, United States; 2NYU School of Medicine, New York, New York, United States. Contact e-mail: [email protected] Background: Currently there is no effective therapy for AD. Immunomodulation stills hold promise but current attempts only address one aspect of the pathology: either amyloid b (Ab) or the hyperphosphorylated tau (ptau) protein. Furthermore current approaches are not specific for the pathological conformers of either protein. We have developed a novel immunomodulatory approach using a pBri peptide. ABri is a rare form of familial human amyloidosis associated with a missense mutation in a stop codon resulting in the transcription of an intronic sequence, leading to production of a highly amyloidogenic protein with a carboxyl terminus that has no sequence homology to any native human protein. We hypothesized that through conformational mimicry the polymerized Bri peptide (pBri) could induce a conformation selective immune response that will recognize pathological proteins such as oligomeric tau and Ab. We have tested this approach in APP/PS1, 3xTg and TgSwDI AD mouse models and have documented that use of pBri as an immunogen reduces: amyloid plaques, vascular amyloid deposits and neurofibrillary tangles. Using pBri we have developed monoclonal antibodies (mAbs). Currently we are characterizing one such mAb specific for pathological conformers of Ab and tau. Methods: Monoclonal antibodies were obtained from pBri inoculations of BALBc mice. Positive hybridomas were selected by their shared reactivity against Ab, PHF and PrP Res. The best conformational candidate (TAB1) was selected after characterization by blots, ELISA and histology against pathological conformers in AD tissue. TAB1 is being used of treatment by intraperitoneal injection in 3xTg mice with both tau and Ab related pathology. Results: TAB1 on tissue sections specifically immunolabels AD tissue with less immunoreactivity on age matched tissue. On Western blots TAB1 detects purified paired helical filament preparations, aggregated/oligomeric Ab and PrP Res, extracted from CJD brain tissue. Use of TAB1 therapeutically in 3xTg mice is on-going. Conclusions: We have developed a novel immunization procedure which we have used to produce monoclonal antibodies (mAbs) that recognize multiple pathological proteins, including PrP Res, oligomer A b and ptau. We are characterizing one of these mAbs, TAB1,
P237
which gives selective immunolabeling in AD tissue and on Western blots to pathological conformers. We believe that immunotherapy that specifically targets the most toxic, oligomeric forms of Ab and ptau, has a greater chance of success with much less risk of toxicity. O3-14-03
CHOLESTEROL AND TAU PATHOLOGY: CONSEQUENCES OF AAV-MEDIATED CHOLESTEROL-24-HYDROXYLASE OVEREXPRESSION IN THE THY-TAU22 MOUSE
Jerome Braudeau1, Marie-Anne Burlot1, Sophie Ayciriex2, Jennifer Varin3, Benoit Gautier4, Fathia Djelti1, Luce Dauphinot5, Zommer Nadege6, Rapha€elle Caillierez6, Laurent Pradier7, Olivier Laprevote8, Ivan Bieche3, Nicolas Auzeil2, Marie-Claude Potier9, Patrick Aubourg1, Luc Buee10, David Blum6, Nathalie Cartier1, 1INSERM UMR 986, Fontenay-aux-Roses, France; 2Faculte de Pharmacie EA4463, Paris, France, Metropolitan; 3INSERM UMR 745, Paris, France; 4INSERM UMR 986, Paris, France; 5ICM, Paris, France; 6INSERM UMR 837, Lille, France; 7Sanofi-Aventis, Vitry sur Seine Cedex, France; 8Faculte de Pharmacie EA4463, Paris, France; 9ICM, Paris, France; 10Universite Lille 2-Inserm UMR 837, Lille, France. Contact e-mail: marieanne.burlot@ gmail.com Background: Emerging evidences suggest a link between brain cholesterol homeostasis and Alzheimer disease (AD), particularly in the production of amyloid Aß peptides from amyloid protein precursor (APP). However, a link between cholesterol homeostasis and AD-like Tau pathology has not yet been established. In the brain, cholesterol is synthesized in situ but cannot be degraded nor cross the blood brain barrier. The major exportable form of brain cholesterol is 24-hydroxycholesterol (24-OHC) generated by the neuronal cholesterol 24-hydroxylase enzyme (CYP46A1). Methods: The THY-Tau22 mouse exhibits progressive neuron-specific AD-like Tau pathology, notably in hippocampus, associated with behavioral impairments. This model was used to study the link between tau pathology and cholesterol metabolism. CYP46A1 overexpression was induced by stereotactic injection of an adeno-associated virus (AAV) vector carrying the human CYP46A1 gene in hippocampus of WT and THY-Tau22 mice. AAV vector coding a mutated inactive CYP46A1 enzyme was used as control vector. Animals were injected at 3.5 months. Behavioral studies (Y maze and MWM) were performed at 6 and 9 months respectively before sacrifice. Samples were analyzed in lipidomics, molecular biology, biochemistry and immunohistochemistry. Results: In this model, we showed a gene expression decrease of enzymes involved in the cholesterol biosynthesis and particularly in the mevalonate pathway. Moreover, CYP46A1 enzyme expression was markedly reduced in the hippocampus. This decrease was associated with a reduction of 24-OHC content. In order to highlight the close relationship between CYP46A1 alteration and THYTau22 phenotype, we overexpressed CYP46A1. In THY-Tau22 mice, CYP46A1 overexpression restored 24-OHC concentration, mevalonate pathway alterations and rescued memory deficits observed. In THY-Tau22 mice, we also characterized an alteration in immediate early gene expressions which are required during memory processes. This decrease is also restored with CYP46A1 overexpression. Analysis of microarray and cellular mechanisms underlying the protective effect of CYP46A1 overexpression on memory deficits observed in THY-Tau22 mouse will be presented. Conclusions: These results are of major importance to get further mechanistic comprehension of existing interaction between cholesterol and Tau pathology in AD and other tauopathies and could be used to define new therapeutic strategies. O3-14-04
UNRAVELLING THE ROLE OF CASEIN KINASE 1 DELTA/EPSILON INHIBITORS IN MODULATING TAU PHOSPHORYLATION AND CELL SIGNALLING PATHWAYS USING GLOBAL PHOSPHOPROTEOMICS IN A TRANSGENIC TAUOPATHY MODEL OF ALZHEIMER’S DISEASE
Ian Pike1, Emma Lahert1, Claire Russell1, Vikram Mitra2, Malcolm Andrew Ward3, 1Proteome Sciences plc, London, England, United
Background: Pathological tau aggregation leading to filamentous tau inclusions characterizes neurodegenerative tauopathies such as Alzheimer’s Disease and frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17). Tau aggregation is linked with clinical symptoms and probably represents the mediator of neurodegeneration. At the moment there is no cure for these diseases and existing treatments are only of palliative nature. Methods: Transgenic mice that overexpress human P301S tau mutation resemble many neuropathological features of human tauopathies in combination with behavioral deficits and increased mortality. We tested the effect of the small molecule anle138b as an anti-aggregative drug on the neuropathology and behavioral deficits in P301S mice. The compound was administered from weaning until death. Results: Here we show that reducing the amount of tau aggregates using the compound anle138b ameliorates disease symptoms in a mouse model for tauopathies. Treatment with anle138b significantly prolonged the survival of transgenic mice and rescued behavioral deficits as well as synapse and neuron loss in the CA3 region and stratum lucidum of the hippocampus. Our results indicate that reducing tau pathology with the anti-aggregative compound anle138b represents a clinically effective and promising approach for the treatment of human tauopathies. Conclusions: In conclusion, anle138b mitigate tau pathology and may represent a new lead compound for developing a therapy for human tauopathies. O3-14-02
TARGETING THE SHARED PATHOLOGICAL CONFORMERS OF BOTH Ab AND HYPERPHOSPHYLATED TAU WITH A CONFORMATIONALLY SELECTIVE MONOCLONAL ANTIBODY
Thomas Wisniewski1, Eleanor Drummond2, Krystal Herline2, Yanjie Sun2, Fernando Goni2, 1New York University School of Medicine, New York, New York, United States; 2NYU School of Medicine, New York, New York, United States. Contact e-mail: [email protected] Background: Currently there is no effective therapy for AD. Immunomodulation stills hold promise but current attempts only address one aspect of the pathology: either amyloid b (Ab) or the hyperphosphorylated tau (ptau) protein. Furthermore current approaches are not specific for the pathological conformers of either protein. We have developed a novel immunomodulatory approach using a pBri peptide. ABri is a rare form of familial human amyloidosis associated with a missense mutation in a stop codon resulting in the transcription of an intronic sequence, leading to production of a highly amyloidogenic protein with a carboxyl terminus that has no sequence homology to any native human protein. We hypothesized that through conformational mimicry the polymerized Bri peptide (pBri) could induce a conformation selective immune response that will recognize pathological proteins such as oligomeric tau and Ab. We have tested this approach in APP/PS1, 3xTg and TgSwDI AD mouse models and have documented that use of pBri as an immunogen reduces: amyloid plaques, vascular amyloid deposits and neurofibrillary tangles. Using pBri we have developed monoclonal antibodies (mAbs). Currently we are characterizing one such mAb specific for pathological conformers of Ab and tau. Methods: Monoclonal antibodies were obtained from pBri inoculations of BALBc mice. Positive hybridomas were selected by their shared reactivity against Ab, PHF and PrP Res. The best conformational candidate (TAB1) was selected after characterization by blots, ELISA and histology against pathological conformers in AD tissue. TAB1 is being used of treatment by intraperitoneal injection in 3xTg mice with both tau and Ab related pathology. Results: TAB1 on tissue sections specifically immunolabels AD tissue with less immunoreactivity on age matched tissue. On Western blots TAB1 detects purified paired helical filament preparations, aggregated/oligomeric Ab and PrP Res, extracted from CJD brain tissue. Use of TAB1 therapeutically in 3xTg mice is on-going. Conclusions: We have developed a novel immunization procedure which we have used to produce monoclonal antibodies (mAbs) that recognize multiple pathological proteins, including PrP Res, oligomer A b and ptau. We are characterizing one of these mAbs, TAB1,
P237
which gives selective immunolabeling in AD tissue and on Western blots to pathological conformers. We believe that immunotherapy that specifically targets the most toxic, oligomeric forms of Ab and ptau, has a greater chance of success with much less risk of toxicity. O3-14-03
CHOLESTEROL AND TAU PATHOLOGY: CONSEQUENCES OF AAV-MEDIATED CHOLESTEROL-24-HYDROXYLASE OVEREXPRESSION IN THE THY-TAU22 MOUSE
Jerome Braudeau1, Marie-Anne Burlot1, Sophie Ayciriex2, Jennifer Varin3, Benoit Gautier4, Fathia Djelti1, Luce Dauphinot5, Zommer Nadege6, Rapha€elle Caillierez6, Laurent Pradier7, Olivier Laprevote8, Ivan Bieche3, Nicolas Auzeil2, Marie-Claude Potier9, Patrick Aubourg1, Luc Buee10, David Blum6, Nathalie Cartier1, 1INSERM UMR 986, Fontenay-aux-Roses, France; 2Faculte de Pharmacie EA4463, Paris, France, Metropolitan; 3INSERM UMR 745, Paris, France; 4INSERM UMR 986, Paris, France; 5ICM, Paris, France; 6INSERM UMR 837, Lille, France; 7Sanofi-Aventis, Vitry sur Seine Cedex, France; 8Faculte de Pharmacie EA4463, Paris, France; 9ICM, Paris, France; 10Universite Lille 2-Inserm UMR 837, Lille, France. Contact e-mail: marieanne.burlot@ gmail.com Background: Emerging evidences suggest a link between brain cholesterol homeostasis and Alzheimer disease (AD), particularly in the production of amyloid Aß peptides from amyloid protein precursor (APP). However, a link between cholesterol homeostasis and AD-like Tau pathology has not yet been established. In the brain, cholesterol is synthesized in situ but cannot be degraded nor cross the blood brain barrier. The major exportable form of brain cholesterol is 24-hydroxycholesterol (24-OHC) generated by the neuronal cholesterol 24-hydroxylase enzyme (CYP46A1). Methods: The THY-Tau22 mouse exhibits progressive neuron-specific AD-like Tau pathology, notably in hippocampus, associated with behavioral impairments. This model was used to study the link between tau pathology and cholesterol metabolism. CYP46A1 overexpression was induced by stereotactic injection of an adeno-associated virus (AAV) vector carrying the human CYP46A1 gene in hippocampus of WT and THY-Tau22 mice. AAV vector coding a mutated inactive CYP46A1 enzyme was used as control vector. Animals were injected at 3.5 months. Behavioral studies (Y maze and MWM) were performed at 6 and 9 months respectively before sacrifice. Samples were analyzed in lipidomics, molecular biology, biochemistry and immunohistochemistry. Results: In this model, we showed a gene expression decrease of enzymes involved in the cholesterol biosynthesis and particularly in the mevalonate pathway. Moreover, CYP46A1 enzyme expression was markedly reduced in the hippocampus. This decrease was associated with a reduction of 24-OHC content. In order to highlight the close relationship between CYP46A1 alteration and THYTau22 phenotype, we overexpressed CYP46A1. In THY-Tau22 mice, CYP46A1 overexpression restored 24-OHC concentration, mevalonate pathway alterations and rescued memory deficits observed. In THY-Tau22 mice, we also characterized an alteration in immediate early gene expressions which are required during memory processes. This decrease is also restored with CYP46A1 overexpression. Analysis of microarray and cellular mechanisms underlying the protective effect of CYP46A1 overexpression on memory deficits observed in THY-Tau22 mouse will be presented. Conclusions: These results are of major importance to get further mechanistic comprehension of existing interaction between cholesterol and Tau pathology in AD and other tauopathies and could be used to define new therapeutic strategies. O3-14-04
UNRAVELLING THE ROLE OF CASEIN KINASE 1 DELTA/EPSILON INHIBITORS IN MODULATING TAU PHOSPHORYLATION AND CELL SIGNALLING PATHWAYS USING GLOBAL PHOSPHOPROTEOMICS IN A TRANSGENIC TAUOPATHY MODEL OF ALZHEIMER’S DISEASE
Ian Pike1, Emma Lahert1, Claire Russell1, Vikram Mitra2, Malcolm Andrew Ward3, 1Proteome Sciences plc, London, England, United