- Email: [email protected]
MULTITEP PLATFORM-BASED DNA VACCINES FOR ALPHA-SYNUCLEINOPATHIES: PRECLINICAL EVALUATION OF IMMUNOGENICITY AND THERAPEUTIC POTENCY
Poster Presentations: Sunday, July 16, 2017
P1-101
MULTITEP PLATFORM-BASED DNA VACCINES FOR ALPHASYNUCLEINOPATHIES: PRECLINICAL EVALUATION OF IMMUNOGENICITY AND THERAPEUTIC POTENCY
David H. Cribbs, University of California Irvine, Irvine, CA, USA. Contact e-mail: [email protected] Background: We have previously demonstrated that anti-Ab DNAvaccine (AV-1959D) based on our proprietary MultiTEP platform technology is extremely immunogenic in mice, rabbits, and monkeys. Importantly, MultiTEP platform enables development of vaccines targeting pathological molecules involved in various neurodegenerative disorders. Taking advantage of the universality of MultiTEP platform, we developed DNA vaccines targeting three B cell epitopes (a85-99, aa109-126, aa 126-140) of human alpha-Synuclein (ha-Syn) separately, or all three epitopes simultaneously. Methods: Taking advantage of the universality of MultiTEP platform, we developed DNAvaccines targeting three B cell epitopes (a85-99, aa109-126, aa 126-140) of human alpha-Synuclein (ha-Syn) separately, or all three epitopes simultaneously. Results: All four DNA vaccines (i) generate high titers of anti-ha-Syn antibodies; (ii) induce robust MultiTEP-specific Th cell responses without activation of potentially detrimental autoreactive anti-ha-Syn Th cells. Generated antibodies recognize misfolded haSyn produced by neuroblastoma cells, ha-Syn in the brain tissues of transgenic mouse strains and in the brain tissues of Dementia with Lewy Bodies cases. Conclusions:Based on these results, the most promising vaccine targeting three B cell epitopes of ha-Syn simultaneously (PV-1950D) has been chosen for ongoing pre-clinical assessment in mouse models of ha-Syn with the aim to translate it to the human clinical trials.
P1-102
HEMODYNAMIC CONNECTIVITY MEASUREMENTS FOLLOW NEURONAL CONNECTIVITY CHANGES IN A MOUSE MODEL OF AMYLOIDOSIS: CONSIDERATIONS FOR FUNCTIONAL CONNECTIVITY MRI
Alberto L. Vazquez1, Mohammed U. Nisar1, Chester Mathis1, Bill E. Klunk1,2, 1University of Pittsburgh, Pittsburgh, PA, USA; 2 Alzheimer’s Disease Research Center, Pittsburgh, PA, USA. Contact e-mail: [email protected] Background: Functional connectivity MRI (fcMRI) has emerged as a technique with early AD detection potential via reductions in the size and strength of connectivity between specific brain areas. However, the hemodynamic nature of fcMRI may reflect changes in vascular dysfunction instead of neuronal dysfunction. A recent study used a mouse model of amyloidosis to show changes in connectivity similar to those measured in AD subjects. This work used the same mouse model along with a genetically-encoded calcium indicator (GCaMP) to directly track neuronal activity in several brain areas and determine the relationship between neuronal connectivity, hemodynamic connectivity (analogous to fcMRI) and amyloid plaque load. Methods: APP-PS1 mice (n¼3) obtained from the MMRRC were injected with AAV-Syn-GCaMP6f virus in retrosplenial, visual and somatosensory cortex. A head plate and cover glass were installed on the skull for longitudinal head-fixed awake imaging (Fig. 1). Images sensitive to neuronal activity (GCaMP) and hemodynamic changes (analogous to fcMRI) were simultaneously acquired for at least 3 months using a
P279
custom imaging system (Fig. 2). Amyloid load was assessed every 6-8 weeks using Methoxy-XO4. The same procedure was performed in control mice (n¼2) as well as APP-PS1 mice injected with vehicle solution (PBS; n¼2). Results: Measurements of neuronal and hemodynamic connectivity between the same brain regions were significantly correlated, in agreement with previous findings from our group. Overall decreases in neuronal connectivity were observed as a function of age when using the retrosplenial cortex as seed region (Fig. 3). Increases in amyloid load were also observed in APP-PS1 mice, as expected (Fig. 4). This study is currently ongoing. Conclusions: Our findings thus far suggest that hemodynamic changes, though less sensitive, reflect changes in neuronal connectivity.
P1-101
MULTITEP PLATFORM-BASED DNA VACCINES FOR ALPHASYNUCLEINOPATHIES: PRECLINICAL EVALUATION OF IMMUNOGENICITY AND THERAPEUTIC POTENCY
David H. Cribbs, University of California Irvine, Irvine, CA, USA. Contact e-mail: [email protected] Background: We have previously demonstrated that anti-Ab DNAvaccine (AV-1959D) based on our proprietary MultiTEP platform technology is extremely immunogenic in mice, rabbits, and monkeys. Importantly, MultiTEP platform enables development of vaccines targeting pathological molecules involved in various neurodegenerative disorders. Taking advantage of the universality of MultiTEP platform, we developed DNA vaccines targeting three B cell epitopes (a85-99, aa109-126, aa 126-140) of human alpha-Synuclein (ha-Syn) separately, or all three epitopes simultaneously. Methods: Taking advantage of the universality of MultiTEP platform, we developed DNAvaccines targeting three B cell epitopes (a85-99, aa109-126, aa 126-140) of human alpha-Synuclein (ha-Syn) separately, or all three epitopes simultaneously. Results: All four DNA vaccines (i) generate high titers of anti-ha-Syn antibodies; (ii) induce robust MultiTEP-specific Th cell responses without activation of potentially detrimental autoreactive anti-ha-Syn Th cells. Generated antibodies recognize misfolded haSyn produced by neuroblastoma cells, ha-Syn in the brain tissues of transgenic mouse strains and in the brain tissues of Dementia with Lewy Bodies cases. Conclusions:Based on these results, the most promising vaccine targeting three B cell epitopes of ha-Syn simultaneously (PV-1950D) has been chosen for ongoing pre-clinical assessment in mouse models of ha-Syn with the aim to translate it to the human clinical trials.
P1-102
HEMODYNAMIC CONNECTIVITY MEASUREMENTS FOLLOW NEURONAL CONNECTIVITY CHANGES IN A MOUSE MODEL OF AMYLOIDOSIS: CONSIDERATIONS FOR FUNCTIONAL CONNECTIVITY MRI
Alberto L. Vazquez1, Mohammed U. Nisar1, Chester Mathis1, Bill E. Klunk1,2, 1University of Pittsburgh, Pittsburgh, PA, USA; 2 Alzheimer’s Disease Research Center, Pittsburgh, PA, USA. Contact e-mail: [email protected] Background: Functional connectivity MRI (fcMRI) has emerged as a technique with early AD detection potential via reductions in the size and strength of connectivity between specific brain areas. However, the hemodynamic nature of fcMRI may reflect changes in vascular dysfunction instead of neuronal dysfunction. A recent study used a mouse model of amyloidosis to show changes in connectivity similar to those measured in AD subjects. This work used the same mouse model along with a genetically-encoded calcium indicator (GCaMP) to directly track neuronal activity in several brain areas and determine the relationship between neuronal connectivity, hemodynamic connectivity (analogous to fcMRI) and amyloid plaque load. Methods: APP-PS1 mice (n¼3) obtained from the MMRRC were injected with AAV-Syn-GCaMP6f virus in retrosplenial, visual and somatosensory cortex. A head plate and cover glass were installed on the skull for longitudinal head-fixed awake imaging (Fig. 1). Images sensitive to neuronal activity (GCaMP) and hemodynamic changes (analogous to fcMRI) were simultaneously acquired for at least 3 months using a
P279
custom imaging system (Fig. 2). Amyloid load was assessed every 6-8 weeks using Methoxy-XO4. The same procedure was performed in control mice (n¼2) as well as APP-PS1 mice injected with vehicle solution (PBS; n¼2). Results: Measurements of neuronal and hemodynamic connectivity between the same brain regions were significantly correlated, in agreement with previous findings from our group. Overall decreases in neuronal connectivity were observed as a function of age when using the retrosplenial cortex as seed region (Fig. 3). Increases in amyloid load were also observed in APP-PS1 mice, as expected (Fig. 4). This study is currently ongoing. Conclusions: Our findings thus far suggest that hemodynamic changes, though less sensitive, reflect changes in neuronal connectivity.