- Email: [email protected]
COMBINATION THERAPY WITH A PLAQUE-SPECIFIC ABETA ANTIBODY AND A BACE INHIBITOR RESULTS IN DRAMATIC PLAQUE REDUCTION IN A DOSE-DEPENDENT MANNER IN AGED PDAPP TRANSGENIC MICE
Poster Presentations: Wednesday, July 27, 2016
from PBMC following the third vaccination, and will be further followed. Results: High antibody responses were found in both species. In the rabbit, 277.1 6 118.4 mg/ml plasma were measured after 5 immunizations with no significant differences between the dose groups. Antibody levels declined slightly to 246.6 6 109.5 mg/ml after a resting period of two months. Low levels of IFNg and IL-17 secreting splenocytes were found in the ELISPOT assays from rabbit splenocytes. Rhesus monkeys reached mean antibody levels of 114.2 6 41.67 mg/ml plasma after five immunizations. The isotype profile was high on IgG4 antibodies, indicative of a Th2 immune response. After three immunizations, no IL-17 or IFNg producing cells were found in ELISPOT assays from PBMCs. Conclusions: DNA Ab42 immunization leads to high antibody titers in large mammals, and is likely to produce high antibody levels and a safe (Th2 biased) immune response in humans as well. supported by NIA/NIH P30AG12300-21, Zale Foundation, Rudman Foundation, AWARE, Presbyterian Village North, Freiberger, and Denker Family Funds
P1191
Front in Cell Neurosci(2015). 5 Jaunmuktane, Z. et al. Nature(2015). 6 Ye, L. et al. Brain Pathol(2015). 7 Patel, K. R. Manag Care (2015). P4-401
COMBINATION THERAPY WITH A PLAQUESPECIFIC ABETA ANTIBODY AND A BACE INHIBITOR RESULTS IN DRAMATIC PLAQUE REDUCTION IN A DOSE-DEPENDENT MANNER IN AGED PDAPP TRANSGENIC MICE
Justin T. Hole1, Margaret M. Racke1, John A. Tzaferis1, Feng Liu1, Herold Oluoch1, Zeshan Ahmed2, Alice Fisher2, Wesley Anderson1, Philip Iversent1, Celedon Gonzales1, Zhixiang Yang1, Leonard N. Boggs1, Scott A. Monk1, Dustin J. Mergott1, Ying YT. Yang3, Jirong Lu1, Michael C. Irizarry1, John R. Sims1, Patrick C. May1, Michael Hutton2, Christer Nordstedt1, Ronald B. DeMattos1, 1Eli Lilly and Company, Indianapolis, IN, USA; 2Eli Lilly and Co., Ltd, Windlesham, United Kingdom; 3Eli Lilly and Company, San Diego, CA, USA. Contact e-mail: [email protected] Background: The use of combination therapies is an emerging and
P4-400
NOVEL AMYLOID-b OLIGOMER-SPECIFIC EPITOPES: A HYPOTHESIS DRIVEN APPROACH TO ALZHEIMER’S IMMUNOTHERAPEUTICS
Judith M. Silverman, Ebrima Gibbs, Jing Wang, Xubiao Peng, Steve Plotkin, Neil R. Cashman, University of British Columbia, Vancouver, BC, Canada. Contact e-mail: [email protected] Background: Alzheimer’s disease (AD) Ab immunotherapeutics in humans have been hampered by negative side-effects including ARIA-E and cerebral microhemorrhage1, mediated by antibody binding to non-toxic fibrils and/or monomers2. Recent advances have demonstrated the fundamental role of amyloid-b oligomers (AbO) in disease onset3, synaptotoxicity4 and progression5,6. Together with promising results from clinical trials targeting aggregated Ab7, these data suggest that by targeting AbOs, a blockade of cognitive decline in humans is possible. We used computational epitope discovery methods to identify immunologic targets on toxic and propagating AbO, that are necessarily absent on Ab monomers and plaques. Methods: Proprietary algorithms have computationally predicted five AbO-specific epitopes. The epitopes were synthesized and conjugated to immunogens, validated for sequence and structure, and used to produce monoclonal antibodies. Surface Plasmon Resonance (SPR) will screen clones against structured and unstructured epitopes and synthetic AbOs, monomers and fibrils. Further validation will include screening antibodies (SPR, immunoprecipitation, immunoblotting) against brain extract and CSF from AD patients and age matched human controls. Antibodies with cross reactivity to monomers and/or fibrils and plaques, determined by immunohistochemistry in brain tissue, will be rejected. Oligomer specific antibodies with no off-target binding will be evaluated in vitrofor the ability to neutralize neurotoxicity and propagation. Results: The target profile of the AbO-specific antibodies is murine monoclonal antibodies that react with synthetic oligomers over monomers with an affinity of least 1/100, or react with native oligomers in brain and CSF with low affinity binding to monomers, and do not react with plaques. Conclusions: The computational epitope discovery approach may produce multiple AbO-specific antibodies. If successful, the approach may be applicable to other proteins and protein misfolding diseases. Acknowledgements: ProMIS Neurosciences, the exclusive licenser of the AbO epitope technology, has supported this work through a sponsored research agreement with UBC.
1 Panza, F. et al. Curr Opin Psychi(2014). 2 Winblad, B. et al. Lancet Neurol(2016). 3 Lesne, S. E. et al. Brain(2013). 4 Ferreira, S. T. et al.
exciting potential path forward for treatment of Alzheimer’s disease (AD). Our understanding of the AD cascade continues to evolve and show the proximal deposition of Ab precedes cognitive impairment by 10 to 15 years and that deposition of Ab has reached a plaque ceiling by the time mild to moderate symptoms manifest. Thus, an increasing belief in the field is that aggressive anti-amyloid interventions, such as combination therapy, may be necessary for treatment of symptomatic Alzheimer’s patients. In order to provide clinically translatable data that moves beyond simple proof-ofconcept, we conducted a large dose response combination therapy study. Methods: Aged PDAPP mice (18-20 months, N ¼ 285) were randomized into an eleven-arm study that consisted of a BACE inhibitor (LY2811376) monotherapy dose response, plaque-specific Ab antibody mE8-IgG2a (anti-Abp3-x) dose response plus highdose BACE inhibitor, high-dose, plaque-specific Ab antibody mE8-IgG2a (anti-Abp3-x) plus BACE inhibitor dose response, and control/time zero cohorts. Animals were sacrificed after 4 months on study and samples were analyzed by various biochemical and immunohistological techniques. Results: We observed a significant dose-response relationship on deposited Ab1-42 (ELISA) for BACE inhibitor monotherapy compared to the control treated animals (-7%, -23%, and -60%). Additionally, histological analyses demonstrated BACE inhibitor treatment significantly lowered the diffuse deposits of Ab. For combination drug treatment arms, we observed a significant broad dose response on deposited Ab1-42 (ELISA) in the presence of high-dose antibody and varying dose levels of BACE inhibitor (from -33% through -84%). The combination drug treatments of a high-dose BACE inhibitor in the presence of varying dose levels of Ab antibody revealed a significant, yet steeper dose response on deposited Ab1-42 (from -60% through -84%). The histological analyses for deposited Ab paralleled observed biochemical changes for combination therapy groups. Several of the of drug combination treatments resulted in significant synergistic lowering of deposited Ab. Conclusions: The combination therapy showed a robust, highly significant and synergistic dose response relationship to plaque lowering in the transgenic mouse model. These results support rationale for using combination therapies in clinical trials.
P4-402
THE IMPACT OF MUSIC-INDUCED AFFECT AND AROUSAL ON OLDER ADULTS’ ATTENTION
Kristen Silveira, Colette M. Smart, Catherine Mateer, University of Victoria, Victoria, BC, Canada. Contact e-mail: [email protected]
from PBMC following the third vaccination, and will be further followed. Results: High antibody responses were found in both species. In the rabbit, 277.1 6 118.4 mg/ml plasma were measured after 5 immunizations with no significant differences between the dose groups. Antibody levels declined slightly to 246.6 6 109.5 mg/ml after a resting period of two months. Low levels of IFNg and IL-17 secreting splenocytes were found in the ELISPOT assays from rabbit splenocytes. Rhesus monkeys reached mean antibody levels of 114.2 6 41.67 mg/ml plasma after five immunizations. The isotype profile was high on IgG4 antibodies, indicative of a Th2 immune response. After three immunizations, no IL-17 or IFNg producing cells were found in ELISPOT assays from PBMCs. Conclusions: DNA Ab42 immunization leads to high antibody titers in large mammals, and is likely to produce high antibody levels and a safe (Th2 biased) immune response in humans as well. supported by NIA/NIH P30AG12300-21, Zale Foundation, Rudman Foundation, AWARE, Presbyterian Village North, Freiberger, and Denker Family Funds
P1191
Front in Cell Neurosci(2015). 5 Jaunmuktane, Z. et al. Nature(2015). 6 Ye, L. et al. Brain Pathol(2015). 7 Patel, K. R. Manag Care (2015). P4-401
COMBINATION THERAPY WITH A PLAQUESPECIFIC ABETA ANTIBODY AND A BACE INHIBITOR RESULTS IN DRAMATIC PLAQUE REDUCTION IN A DOSE-DEPENDENT MANNER IN AGED PDAPP TRANSGENIC MICE
Justin T. Hole1, Margaret M. Racke1, John A. Tzaferis1, Feng Liu1, Herold Oluoch1, Zeshan Ahmed2, Alice Fisher2, Wesley Anderson1, Philip Iversent1, Celedon Gonzales1, Zhixiang Yang1, Leonard N. Boggs1, Scott A. Monk1, Dustin J. Mergott1, Ying YT. Yang3, Jirong Lu1, Michael C. Irizarry1, John R. Sims1, Patrick C. May1, Michael Hutton2, Christer Nordstedt1, Ronald B. DeMattos1, 1Eli Lilly and Company, Indianapolis, IN, USA; 2Eli Lilly and Co., Ltd, Windlesham, United Kingdom; 3Eli Lilly and Company, San Diego, CA, USA. Contact e-mail: [email protected] Background: The use of combination therapies is an emerging and
P4-400
NOVEL AMYLOID-b OLIGOMER-SPECIFIC EPITOPES: A HYPOTHESIS DRIVEN APPROACH TO ALZHEIMER’S IMMUNOTHERAPEUTICS
Judith M. Silverman, Ebrima Gibbs, Jing Wang, Xubiao Peng, Steve Plotkin, Neil R. Cashman, University of British Columbia, Vancouver, BC, Canada. Contact e-mail: [email protected] Background: Alzheimer’s disease (AD) Ab immunotherapeutics in humans have been hampered by negative side-effects including ARIA-E and cerebral microhemorrhage1, mediated by antibody binding to non-toxic fibrils and/or monomers2. Recent advances have demonstrated the fundamental role of amyloid-b oligomers (AbO) in disease onset3, synaptotoxicity4 and progression5,6. Together with promising results from clinical trials targeting aggregated Ab7, these data suggest that by targeting AbOs, a blockade of cognitive decline in humans is possible. We used computational epitope discovery methods to identify immunologic targets on toxic and propagating AbO, that are necessarily absent on Ab monomers and plaques. Methods: Proprietary algorithms have computationally predicted five AbO-specific epitopes. The epitopes were synthesized and conjugated to immunogens, validated for sequence and structure, and used to produce monoclonal antibodies. Surface Plasmon Resonance (SPR) will screen clones against structured and unstructured epitopes and synthetic AbOs, monomers and fibrils. Further validation will include screening antibodies (SPR, immunoprecipitation, immunoblotting) against brain extract and CSF from AD patients and age matched human controls. Antibodies with cross reactivity to monomers and/or fibrils and plaques, determined by immunohistochemistry in brain tissue, will be rejected. Oligomer specific antibodies with no off-target binding will be evaluated in vitrofor the ability to neutralize neurotoxicity and propagation. Results: The target profile of the AbO-specific antibodies is murine monoclonal antibodies that react with synthetic oligomers over monomers with an affinity of least 1/100, or react with native oligomers in brain and CSF with low affinity binding to monomers, and do not react with plaques. Conclusions: The computational epitope discovery approach may produce multiple AbO-specific antibodies. If successful, the approach may be applicable to other proteins and protein misfolding diseases. Acknowledgements: ProMIS Neurosciences, the exclusive licenser of the AbO epitope technology, has supported this work through a sponsored research agreement with UBC.
1 Panza, F. et al. Curr Opin Psychi(2014). 2 Winblad, B. et al. Lancet Neurol(2016). 3 Lesne, S. E. et al. Brain(2013). 4 Ferreira, S. T. et al.
exciting potential path forward for treatment of Alzheimer’s disease (AD). Our understanding of the AD cascade continues to evolve and show the proximal deposition of Ab precedes cognitive impairment by 10 to 15 years and that deposition of Ab has reached a plaque ceiling by the time mild to moderate symptoms manifest. Thus, an increasing belief in the field is that aggressive anti-amyloid interventions, such as combination therapy, may be necessary for treatment of symptomatic Alzheimer’s patients. In order to provide clinically translatable data that moves beyond simple proof-ofconcept, we conducted a large dose response combination therapy study. Methods: Aged PDAPP mice (18-20 months, N ¼ 285) were randomized into an eleven-arm study that consisted of a BACE inhibitor (LY2811376) monotherapy dose response, plaque-specific Ab antibody mE8-IgG2a (anti-Abp3-x) dose response plus highdose BACE inhibitor, high-dose, plaque-specific Ab antibody mE8-IgG2a (anti-Abp3-x) plus BACE inhibitor dose response, and control/time zero cohorts. Animals were sacrificed after 4 months on study and samples were analyzed by various biochemical and immunohistological techniques. Results: We observed a significant dose-response relationship on deposited Ab1-42 (ELISA) for BACE inhibitor monotherapy compared to the control treated animals (-7%, -23%, and -60%). Additionally, histological analyses demonstrated BACE inhibitor treatment significantly lowered the diffuse deposits of Ab. For combination drug treatment arms, we observed a significant broad dose response on deposited Ab1-42 (ELISA) in the presence of high-dose antibody and varying dose levels of BACE inhibitor (from -33% through -84%). The combination drug treatments of a high-dose BACE inhibitor in the presence of varying dose levels of Ab antibody revealed a significant, yet steeper dose response on deposited Ab1-42 (from -60% through -84%). The histological analyses for deposited Ab paralleled observed biochemical changes for combination therapy groups. Several of the of drug combination treatments resulted in significant synergistic lowering of deposited Ab. Conclusions: The combination therapy showed a robust, highly significant and synergistic dose response relationship to plaque lowering in the transgenic mouse model. These results support rationale for using combination therapies in clinical trials.
P4-402
THE IMPACT OF MUSIC-INDUCED AFFECT AND AROUSAL ON OLDER ADULTS’ ATTENTION
Kristen Silveira, Colette M. Smart, Catherine Mateer, University of Victoria, Victoria, BC, Canada. Contact e-mail: [email protected]