The multi-target effects of CNI-1493: convergence of anti-amylodogenic and anti-inflammatory properties in animal models of Alzheimer's disease.

After several decades of Alzheimer's disease (AD) research and failed clinical trials, one can speculate that targeting a single pathway is not sufficient. However, a cocktail of novel therapeutics will constitute a challenging clinical trial. A more plausible approach will capitalize on a drug that has relevant and synergistic multiple-target effects in AD. We have previously demonstrated the efficacy of CNI-1493 in the CRND8 transgenic AD mouse model. Similar to many anti-inflammatory drugs that were tested in preclinical model of AD, it was speculated that the significant effect of CNI-1493 is due to its established anti-inflammatory properties in rodents and humans. In the present study, we set out to elucidate the protective mechanism of CNI-1493 as a drug simultaneously targeting several aspects of AD pathology. Using C1213, a highly similar analogue of CNI-1493 that lacks anti-inflammatory properties, we show that both compounds directly interact with soluble and insoluble Amyloid ß (Aß) aggregates and attenuate Aß cytotoxicity in vitro. Additionally, CNI-1493 and C1213 ameliorated Aß-induced behavioral deficits in nematodes. Finally, C1213 reduced Aß plaque burden and cognitive deficits in transgenic CRND8 mice to a similar extent as previously shown with CNI-1493. Taken together, our findings suggest anti-amyloidogenic activity as a relevant component for the in-vivo efficacy of CNI-1493 and its analogue C1213. Thus, CNI-1493, a drug with proven safety in humans, is a viable candidate for novel multi-target therapeutic approaches to AD.

Mechanisms of action of naturally occurring antibodies against ß-amyloid on microglia.

BACKGROUND: Naturally occurring autoantibodies against amyloid-ß (nAbs-Aß) have been shown to exert beneficial effects on transgenic Alzheimer's disease (AD) animals in vivo and on primary neurons in vitro. Not much is known about their effect on microglial cells. Our aim was to investigate the effect of nAbs-Aß on amyloid-ß (Aß)-treated microglial cells in vitro with respect to cell viability, stress pathways, cytokine production and phagocytotic abilities and whether these effects can be conveyed to neurons. METHODS: Primary microglial cells isolated from Swiss Webster mouse mesencephalons on embryonic day 13.5 were pretreated with nAbs-Aß and then treated with Aß oligomers. After 3 hours, phagocytosis as well as western blot analysis were evaluated to measure the amount of phagocytized Aß. Cell viability was analyzed using an MTT assay 24 hours after treatment. Pro-inflammatory cytokines in the supernatants were analyzed with ELISAs and then we treated primary neuronal cells with these conditioned microglia supernatants. Twenty-four hours later we did a MTT assay of the treated neurons. We further investigated the effect of a single nAbs-Aß administration on Tg2576 mice in vivo. RESULTS: Upon co-administration of Aß and nAbs-Aß no change in microglia viability was observed. However, there was an increase in phosphorylated p38 protein level, an increase in the pro-inflammatory cytokines TNF-α and IL-6 and an increase in Aß uptake by microglial cells. Treatment of primary neurons with conditioned microglia medium led to a 10% improvement in cell viability when nAbs-Aß were co-administered compared to Aß-treated cells alone. We were unable to detect changes in cytokine production in brain lysates of Tg2576 mice. CONCLUSIONS: We provide evidence on the mechanism of action of nAbs-Aß on microglia in vitro. Interestingly, our in vivo data indicate that nAbs-Aß administration should be considered as a therapeutic strategy in AD, since there is no inflammatory reaction.

Naturally occurring autoantibodies against beta-amyloid: investigating their role in transgenic animal and in vitro models of Alzheimer's disease.

Alzheimer's disease (AD) is a neurodegenerative disorder primarily affecting regions of the brain responsible for higher cognitive functions. Immunization against ß-amyloid (Aß) in animal models of AD has been shown to be effective on the molecular level but also on the behavioral level. Recently, we reported naturally occurring autoantibodies against Aß (NAbs-Aß) being reduced in Alzheimer's disease patients. Here, we further investigated their physiological role: in epitope mapping studies, NAbs-Aß recognized the mid-/C-terminal end of Aß and preferentially bound to oligomers but failed to bind to monomers/fibrils. NAbs-Aß were able to interfere with Aß peptide toxicity, but NAbs-Aß did not readily clear senile plaques although early fleecy-like plaques were reduced. Administration of NAbs-Aß in transgenic mice improved the object location memory significantly, almost reaching performance levels of wild-type control mice. These findings suggest a novel physiological mechanism involving NAbs-Aß to dispose of proteins or peptides that are prone to forming toxic aggregates.

APP transgenic mice: the effect of active and passive immunotherapy in cognitive tasks.

Various immunotherapy strategies for APP transgenic mice have emerged in recent years. Specifically, active immunization with beta-amyloid (A beta) or passive immunization with anti-A beta-antibodies in APP transgenic mice has appeared most promising. Recent studies have shown that treatment of APP transgenic mice either with A beta(40/42) or A beta-specific antibodies can have beneficial effects in cognitive tasks. Active as well as passive immunization have been shown to affect spatial, non-spatial, emotional and object-related learning and memory. Such effects can be observed when treatments are applied prophylactically (before apparent A beta pathology) or therapeutically (after the development of A beta pathology) in APP transgenic mice. This review focuses on such cognitive outcomes of different active and passive immunization strategies in APP transgenic mice.

Effects of neonatal and peripubertal ethanol treatment on various aspects of adult rat behavior and brain anatomy.

Exposure to ethanol during critical stages of brain development and maturation has adverse effects on behavioral and cognitive functions. So far, most animal models focused on the effects of either pre- or early postnatal ethanol treatment on behavior. We here used a multiple crossover design to investigate the effects of neonatal (postnatal day 7) ethanol treatment (2.5 g/kg b.i.d., dissolved in saline), subchronic peripubertal (postnatal days 40-65) ethanol treatment (1.0 g/kg, dissolved in saline) and the combination of both on the performance of adult Wistar rats in a variety of behavioral tasks. We also assessed anatomical changes in limbic and cortical brain areas. No effects of either single or combined neonatal and pubertal ethanol treatment was found on prepulse inhibition of startle (PPI, a measure of sensorimotor gating), or on the acoustic startle response in the absence of prepulses. Peripubertal ethanol treatment reduced the explorative behavior in the open field. The breakpoint in a progressive ratio operant response task was increased in those rats that had received both neonatal and pubertal ethanol treatment, while the preference for palatable food used as reinforcer in this task was not affected. No treatment effects were found on object recognition memory. No treatment effects on anxiety-related behavior in the elevated plus maze were found, however, the anxiolytic effect of the prototypical benzodiazepine diazepam was enhanced in rats that had received peripubertal ethanol treatment. Additive effects of neonatal and pubertal ethanol treatments were found on behaviors related to spontaneous locomotor activity. Combined neonatal and pubertal ethanol treatment lead to a reduction of myelin sheaths in the prefrontal cortex, and the neonatal ethanol treatment lead to a reduced number of parvalbumine-immunoreactive cells in the dorsal hippocampus. These findings suggest that neonatal ethanol exposure increases the risk of some but not all adverse behavioral and brain anatomical effects of pubertal ethanol consumption.

Enhanced prepulse inhibition of startle using salient prepulses in rats.

Prepulse inhibition (PPI) of the startle reflex occurs when a non-startling stimulus is presented shortly prior to the startling stimulus. PPI is an operational measure for sensorimotor gating. PPI in humans is enhanced by attention, but there is no evidence yet for attentional modulation of PPI in animals. We here combined PPI and conditioned inhibition paradigms in order to investigate attentional modulation of PPI in rats. PPI was assessed before and after training for conditioned inhibition of fear with the conditioned stimulus (auditory CS) and conditioned inhibitor (visual CI) as prepulses. The CI significantly enhanced PPI after training, whereas presentation of the CS had no effect on PPI. These data suggest attentional modulation of PPI in rats by biologically salient prestimuli. This new paradigm may be useful for examining attentional modulation of PPI in animals and to compare attentional modulation in humans and animals.