MicroRNA silencing: A promising therapy for Alzheimer's disease.

Alzheimer's disease (AD) is a global health crisis currently afflicting ~6 million Americans (and ~40 million people worldwide). By the middle of the century, these numbers will stagger by ~16 million Americans (and ~152 million people worldwide) suffering from AD, if breakthrough disease-modifying treatments are not discovered. Currently, there are no treatments to prevent, halt or cure the disease. Multiple independent studies on brain gene expression patterns have indicated that in AD about 1/3rd of the genes are upregulated while the rest 2/3rd of the genes are downregulated. In that regard, AD therapeutics focused on antagomiR-mediated silencing of"upregulated"microRNAs (miRs) may be more feasible since upregulated miRs in AD continue to increase with the disease progression, as opposed to agomiR-mediated overexpression of down-regulated miRs with unpredictable reduced presence and relative short-life of 1-3h under pathological conditions in AD brain. Studies reported thus far indicate that most of the upregulated pathogenic genes in AD are regulated by pro-inflammatory microRNAs (miRs). Given the precedence of chronic neuroinflammation in triggering AD-like neurodegeneration and multifactorial nature of AD, silencing inflammation-specific micro-RNAs using antisense-microRNAs may be an effective adjuvant therapeutic strategy to prevent, halt or cure AD.

Maternal and Fetal Outcome After Cardiac Operations During Pregnancy: A Meta-Analysis.

BACKGROUND: In the past, cardiac surgical procedures during pregnancy have been associated with a high risk of adverse maternal and fetal outcomes. Therefore, this meta-analysis of more recent studies was done to assess the fetomaternal risk after cardiac operations during pregnancy using cardiopulmonary bypass. METHODS: The MEDLINE, Embase, and Cochrane library were searched to find studies from January 1, 1990, to July 31, 2016, without language restriction. We selected studies that included at least 4 women to report fetomaternal outcomes after a cardiac operation using cardiopulmonary bypass during pregnancy. Two authors independently extracted data from the selected studies. The studies were assessed for methodological qualities using the Newcastle-Ottawa Scale. The primary outcomes included maternal death and any pregnancy loss. The secondary outcomes were maternal complications and neonatal complications. Primary analysis calculated absolute risks and 95% confidence intervals (CIs) for pregnancy outcomes using the DerSimonian-Laird random effects model. Heterogeneity was assessed by I2 statistic and visual plot. RESULTS: Ten studies, including 154 women, were eligible for inclusion in this study. The patients underwent cardiac operations during pregnancy involving cardiopulmonary bypass. As calculated per 100 pregnancies, the pooled unadjusted estimate of maternal mortality was 11.2 (95% CI, 6.8 to 17.8), pregnancy loss was 33.1 (95% CI, 25.1 to 41.2), maternal complications were 8.8 (95% CI, 2.8 to 24.2), and neonatal complications were 10.8 (95% CI, 4.2 to 25.2). The risks of preterm labor and cesarean delivery were 28 per 100 pregnancies (95% CI, 15.6 to 45) and 33.8 per 100 pregnancies (95% CI, 19.1 to 52.4), respectively. CONCLUSIONS: The fetomaternal mortality and morbidity after a cardiac operation during pregnancy are higher than that reported in the earlier literature (PROSPERO No. CRD42016047093).

Anti-edema effects of rhEpo in experimental traumatic brain injury.

PURPOSE: Traumatic brain injury (TBI) is one of the leading causes of disability and death which begins with the formation of edema as the persistent primary causative factor in TBI. Although medical management of cerebral edema by hypothermia, ventriculostomy, mannitol or hypertonic saline have been effective in treating edema, many of these therapies end up with some neurologic deficits, necessitating novel treatment options for treating post-TBI edema. This study investigated edema reducing effects of recombinant human Erythropoietin (rhEPO) in reducing acute brain edema in the CCI mouse model of TBI. METHODS: Anti-edema effects of rhEpo in reducing acute brain edema after injury in the CCI mouse model of TBI were assessed by T2 weighted magnetic resonance imaging (T2wMRI) as the accurate detector of brain edema in correlation with Western blot analysis of cerebral aquaporin 4 (AQP4) index as the critical marker of edema. RESULTS: Results show that rhEpo treatment significantly reduced brain edema with concomitant reduction in AQP4 immunoexpression in the CCI mouse model of TBI. CONCLUSION: Current results emphasize clinical utility of rhEpo in treating post-TBI edema.

Brain Uptake of Neurotherapeutics after Intranasal versus Intraperitoneal Delivery in Mice.

There is a growing global prevalence of neurodegenerative diseases such as Alzheimer's disease and dementia. Current treatment for neurodegenerative diseases is limited due to the blood brain barrier's ability to restrict the entry of therapeutics to the brain. In that context, direct delivery of drugs from nose to brain has gained emerging interest as an important alternative to oral and parenteral routes of administration. Although there are considerable reports showing promising results after intranasal drug delivery in various disease-models and investigatory human clinical trials, there are very few studies showing a detailed pharmacokinetics with regard to the uptake and retention of intranasally delivered material(s) within specific brain regions, which are critical determining factors for dosing conditions and optimal treatment regimen. This investigation compared a time-dependent brain uptake and resident time of various radiolabeled candidate neurotherapeutics after a single bolus intranasal or intraperitoneal administration in mice. Results indicate that the brain uptake of intranasally delivered therapeutic(s) is > 5 times greater than that after intraperitoneal delivery. The peak uptake and resident time of all intranasally delivered test therapeutics for all brain regions is observed to be between 30min-12h, depending upon the distance of brain region from the site of administration, followed by gradual fading of radioactive counts by 24h post intranasal administration. Current study confirms the usefulness of intranasal administration as a non- invasive and efficient means of delivering therapeutics to the brain to treat neurodegenerative diseases including Alzheimer's disease.

Chronic neurodegenerative consequences of traumatic brain injury.

Traumatic brain injury (TBI) is a serious public health concern and a major cause of death and disability worldwide. Each year, an estimated 1.7 million Americans sustain TBI of which ~52,000 people die, ~275,000 people are hospitalized and 1,365,000 people are treated as emergency outpatients. Currently there are ~5.3 million Americans living with TBI. TBI is more of a disease process than of an event that is associated with immediate and long-term sensomotor, psychological and cognitive impairments. TBI is the best known established epigenetic risk factor for later development of neurodegenerative diseases and dementia. People sustaining TBI are ~4 times more likely to develop dementia at a later stage than people without TBI. Single brain injury is linked to later development of symptoms resembling Alzheimer's disease while repetitive brain injuries are linked to later development of chronic traumatic encephalopathy (CTE) and/or Dementia Pugilistica (DP). Furthermore, genetic background of ß-amyloid precursor protein (APP), Apolipoprotein E (ApoE), presenilin (PS) and neprilysin (NEP) genes is associated with exacerbation of neurodegenerative process after TBI. This review encompasses acute effects and chronic neurodegenerative consequences after TBI.

Experimental induction of type 2 diabetes in aging-accelerated mice triggered Alzheimer-like pathology and memory deficits.

Alzheimer's disease (AD) is an age-dependent neurodegenerative disease constituting ~95% of late-onset non-familial/sporadic AD, and only ~5% accounting for early-onset familial AD. Availability of a pertinent model representing sporadic AD is essential for testing candidate therapies. Emerging evidence indicates a causal link between diabetes and AD. People with diabetes are >1.5-fold more likely to develop AD. Senescence-accelerated mouse model (SAMP8) of accelerated aging displays many features occurring early in AD. Given the role played by diabetes in the pre-disposition of AD, and the utility of SAMP8 non-transgenic mouse model of accelerated aging, we examined if high fat diet-induced experimental type 2 diabetes in SAMP8 mice will trigger pathological aging of the brain. Results showed that compared to non-diabetic SAMP8 mice, diabetic SAMP8 mice exhibited increased cerebral amyloid-ß, dysregulated tau-phosphorylating glycogen synthase kinase 3ß, reduced synaptophysin immunoreactivity, and displayed memory deficits, indicating Alzheimer-like changes. High fat diet-induced type 2 diabetic SAMP8 mice may represent the metabolic model of AD.

Brain transit and ameliorative effects of intranasally delivered anti-amyloid-ß oligomer antibody in 5XFAD mice.

Alzheimer's disease (AD) is a global health crisis with limited treatment options. Despite major advances in neurotherapeutics, poor brain penetration due to the blood-brain barrier continues to pose a big challenge in overcoming the access of therapeutics to the central nervous system. In that regard, the non-invasive intranasal route of brain targeting is gaining considerable attention. The nasal mucosa offers a large surface area, rapid absorption, and avoidance of first-pass metabolism increasing drug bioavailability with less systemic side effects. Intranasal delivery is known to utilize olfactory, rostral migratory stream, and trigeminal routes to reach the brain. This investigation confirmed that intranasal delivery of oligomeric amyloid-ß antibody (NU4) utilized all three routes to enter the brain with a resident time of 96 hours post single bolus intranasal administration, and showed evidence of perikaryal and parenchymal uptake of NU4 in 5XFAD mouse brain, confirming the intranasal route as a non-invasive and efficient way of delivering therapeutics to the brain. In addition, this study demonstrated that intranasal delivery of NU4 antibody lowered cerebral amyloid-ß and improved spatial learning in 5XFAD mice.

Wheat germ agglutinin enhanced cerebral uptake of anti-Aß antibody after intranasal administration in 5XFAD mice.

Alzheimer's disease (AD) is the 6th leading cause of death in United States afflicting >5 million Americans. This number is estimated to triple by the middle of the century if effective treatments are not discovered. Current therapy for AD is mainly symptomatic. Effective disease-modifying treatments are needed that would eliminate the cause rather than the symptoms of the disease. Polymerization of monomeric beta-amyloid peptide (Aß) into dimers, soluble oligomers and insoluble fibrils is considered the prime causative factor in triggering AD pathogenesis. Based on these facts, removal/reduction of Aß has gained importance as a primary therapeutic target in treating the cause of the disease. In that regard, passive immunotherapy with direct delivery of anti-Aß antibodies to the brain has shown great promise, but awaits the challenge of overcoming greater influx of anti-Aß antibody into the brain. This investigation was undertaken to maximize direct delivery of immunotherapeutics to the brain by using wheat germ agglutinin (WGA) as a novel axonal transporter-carrier to be conjugated with anti-Aß antibody (6E10) raised against EFRHDS 3-8 amino acid (aa) epitopes of Aß known to react with 1-16 aa residues of mono-/di-/oligomeric Aß. This is the first report showing the use of WGA as an efficient axonal transporter carrier that not only enhanced the influx of anti-Aß antibody directly into the brain but also resulted in greater reduction of cerebral Aß compared to the unconjugated anti-Aß antibody delivered intranasally in Alzheimer's 5XFAD model.

Restoration of cognitive deficits after statin feeding in TBI.

PURPOSE: Traumatic brain injury (TBI) is a global health concern and growing socio-economic burden with limited treatment options. Behavioral assessment in experimental TBI with candidate therapeutic interventions is critical in order to expedite clinical translation. Statins constitute one of the potential treatment options based on their proven beneficial effects in various models of neurotrauma. We compared functional outcome after dietary intervention with representative hydrophilic Pravastatin or lipophilic Simvastatin and Lovastatin to test if different statins will differentially affect cognitive outcomes after injury in a controlled cortical impact injury (CCI) mouse model of TBI. METHODS: Mice were subjected to TBI with a controlled cortical impact produced on the left somatosensory-parietal cortex between bregma -1.82 and -2.06, fed with Simvastatin/Lovastatin/Pravastatin (2 mg/kg) for 8 weeks, evaluated for learning, memory and spontaneous exploration behavior followed by immunocytochemistry of an axonal marker. RESULTS: Results indicate that feeding of TBI mice with Simvastatin and Lovastatin significantly improved spatial learning and memory, restored spontaneous exploration and restored axonal integrity (Simvastatin > Lovastatin). On the other hand, Pravastatin failed to improve spatial learning or memory or exploration or axonal damage. CONCLUSIONS: Current findings confirm maximum benefits rendered by Simvastatin and reinforce Simvastatin as the candidate therapy for treating TBI.

Oxidative insults to neurons and synapse are prevented by aged garlic extract and S-allyl-L-cysteine treatment in the neuronal culture and APP-Tg mouse model.

Alzheimer's disease (AD) is one of the most common forms of dementia in the elderly. In AD patients, ß-amyloid peptide (Aß) plaques and neurofibrillary tangles are common features observed in the CNS. Aß deposition results in the production of reactive oxygen species (ROS) leading to the hyperphosphorylation of tau that are associated with neuronal damage. Cholinesterase inhibitors and a partial NMDA receptor antagonist (memantine) have been identified as potential treatment options for AD. However, clinical studies have found that these drugs fail to prevent the disease progression. From ancient times, garlic (Allium sativum) has been used to treat several diseases. By 'aging' of garlic, some adverse reactions of garlic can be eliminated. Recent findings suggest that 'aged garlic extract' (AGE) may be a therapeutic agent for AD because of its antioxidant and Aß lowering properties. To date, the molecular properties of AGE have been sparsely studied in vitro or in vivo. The present study tested specific biochemical and molecular effects of AGE in neuronal and AD rodent models. Furthermore, we identified S-allyl-L-cysteine (SAC) as one of the most active chemicals responsible for the AGE-mediated effect(s). We observed significant neuroprotective and neurorescue properties of AGE and one of its ingredients, SAC, from ROS (H(2)O(2))-mediated insults to neuronal cells. Treatment of AGE and SAC were found to protect neuronal cells when they were independently co-treated with ROS. Furthermore, a novel neuropreservation effect of AGE was detected in that pre-treatment with AGE alone protected ∼ 80% neuronal cells from ROS-mediated damage. AGE was also found to preserve pre-synaptic protein synaptosomal associated protein of 25 kDa (SNAP25) from ROS-mediated insult. For example, treatment with 2% AGE containing diet and SAC (20 mg/kg of diet) independently increased (∼70%) levels of SNAP25 and synaptophysin in Alzheimer's amyloid precursor protein-transgenic mice, of which the latter was significantly decreased in AD. Taken together, the neuroprotective, including preservation of pre-synaptic proteins by AGE and SAC can be utilized in future drug development in AD.

Synergistic benefits of erythropoietin and simvastatin after traumatic brain injury.

Simvastatin and recombinant human erythropoietin (rhEpo) are implicated as potential therapeutic candidates for traumatic brain injury (TBI). Prominent effects of simvastatin include its anti-inflammatory, neurotrophic and neuroregenerative actions studied in various models of neuronal injury. On the other hand, rhEpo has been shown to promote cell survival mechanisms by producing anti-apoptotic and cell proliferative actions. Beneficial effects of rhEpo and statin monotherapies have been well studied. However, there are no reports showing combined use of rhEpo and statins after TBI. This investigation examined if combined efficacy of cell proliferative ability of rhEpo along with the neuroregenerative ability of simvastatin will render maximum recovery in a controlled cortical impact (CCI) mouse model of TBI. Results showed that compared to baseline TBI, rhEpo was more effective than simvastatin in promoting cell proliferation while simvastatin was more effective than rhEpo in restoring axonal damage following TBI. Combined treatment with simvastatin and rhEpo maximally restored axonal integrity while simultaneously inducing greater proliferation of newly formed cells resulting in better functional recovery after TBI than either alone. This is the first study showing the efficacy of erythropoietin-simvastatin combinational therapeutic approach in achieving greater structural and cognitive recovery after TBI.

Neuroanatomical correlation of behavioral deficits in the CCI model of TBI.

Traumatic brain injury (TBI) is the leading cause of death and disability both in combat and civilian situations with limited treatment options including surgical removal of hematoma, ventricular drainage and use of hyperosmotic agents that restrict secondary injury following TBI. Availability of appropriate model system with full-range characterization of anatomical and behavioral components correlative with brain injury provides a pre-clinical platform to test candidate therapies for clinical translation. Modeling of TBI using controlled cortical impact injury (CCI) is largely considered to be close to clinical TBI and hence CCI models have been widely used in pre-clinical TBI research. Most studies reported so far using CCI models were presented with a limited behavioral characterization and lacked its correlation with the signature histopathology of TBI. Current investigation validated a detailed sensomotor and cognitive behavioral characterization correlative with diffuse axonal injury-the signature histopathology of TBI, in the CCI mouse model of TBI. Present study offers a comprehensively characterized model of TBI that can be used to investigate cellular and molecular mechanisms underlying TBI and to test candidate therapies in developing novel and effective treatments for TBI.

Amelioration of early cognitive deficits by aged garlic extract in Alzheimer's transgenic mice.

Subtle accumulation of beta-amyloid peptide (Abeta) oligomers of Abeta42 species in particular, is known to correlate with cognitive deficits independent of Abeta plaque deposition in the brain. Majority of the research showing behavioral improvement after cerebral Abeta reduction has been reported when the animals carried fewer/abundant amyloid plaques in the brain. Very few studies have addressed whether or not behavioral deficits exist even at the pre-plaque stage or in the absence of plaques that would parallel the mild cognitive impairment (MCI) stage of Alzheimer's disease (AD). Current study was undertaken to determine whether there exists any cognitive impairment during the pre-plaque stage which may parallel the MCI stage of AD, and to confirm whether the observed behavioral deficits correlate with Abeta42 predominance. In addition, the study determined whether anti-amyloidogenic effects of dietary aged garlic extract would prevent progressive behavioral impairment. For this purpose we used Tg2576 model showing slow plaque development with a predominance of Abeta40, and the TgCRND8 model showing accelerated plaque development with a predominance of Abeta42. The results show that at 2 months of age Tg2576 mice did not exhibit behavioral impairment in any of the tasks studied. While 2-month-old TgCRND8 mice displayed only a subtle behavioral deficit that matched the behavioral deficits observed in 7-month-old Tg2576 mice which may correlate with the MCI stage of AD. TgCRND8 mice at 7 months of age exhibited advanced deterioration in all behavioral tasks studied, suggesting that accelerated Abeta accumulation and the predominance of Abeta42 species may account for the pronounced cognitive deficits observed in TgCRND8. Feeding of aged garlic extract prevented deterioration of hippocampal based memory tasks in these mice, suggesting that aged garlic extract has a potential for preventing AD progression.

Antisense inhibition at the beta-secretase-site of beta-amyloid precursor protein reduces cerebral amyloid and acetyl cholinesterase activity in Tg2576.

Misprocessing of beta-amyloid precursor protein (APP) leading to the formation of elevated quantities of beta-amyloid peptide (Abeta), derived by a cleavage at the beta-secretase site (N-671/673aa) and by a cleavage at the gamma-secretase site (C-711/713aa) of APP, is considered a key event in the pathogenesis of Alzheimer disease (AD). Point mutations near the beta-secretase site in the human gene for APP, such as in the Swedish mutation-KM670/671NL, lead to a form of dominantly inherited AD. These mutations are known to promote beta-site cleavage and to increase levels of Abeta. Abeta has been shown previously to increase acetyl cholinesterase (AChE) activity in vitro. We wished to test whether translational blocking of APP-mRNA at the mutated beta-site by antisense (AS) oligodeoxynucleotides (ODNs) directed to the mutated site will reduce cerebral amyloid in the Swedish transgenic mouse model (Tg2576). Mice were injected i.c.v. with AS-ODNs directed at the mutated beta-site (AS-beta site) or with AS-ODNs directed at the normal gamma-site (AS-gamma site) of human APP-mRNA, and compared with procedural controls that received i.c.v. injections of sense ODNs at the beta-site (S-beta site), sense ODNs at the gamma-site (S-gamma site) or mismatched ODNs, and with untreated littermates (Lt) and untreated transgenic mice (Tgs). ODNs were injected into the 3rd ventricle once a week for 4 weeks. Brains were processed for enzyme-linked immunosorbent assay analysis of beta- and gamma-cleaved soluble Abeta40 (sAbeta40), beta- and gamma-cleaved soluble Abeta42 (sAbeta42) and alpha-cleaved soluble beta-amyloid precursor protein (sAPPalpha). The physiological relevance of AS ODNs was tested by evaluating the cerebral distribution of AChE before and after the treatment. AChE was found increased about fivefold in Tg cortex as compared with control brain. Results show that compared with untreated and procedural controls, AS-beta increased cerebral levels of sAPPalpha by 43% and reduced sAbeta40/42 by approximately 39%; while simultaneously reducing the cortical density of AChE by approximately fourfold in the treated Tg animals, almost to the level found in the control brain (all values P<0.0001, analysis of variance, unpaired two-tailed Student's t-test), while AS-gamma did not have any effect. These results indicate that AS directed to the mutated beta-site may be an effective approach to treat familial AD.

Intracerebroventricular passive immunization with anti-oligoAbeta antibody in TgCRND8.

Based on the central dogma of beta-amyloid (Abeta) as a key seeding event in the pathogenesis of Alzheimer disease (AD), immunoneutralization strategies have been actively pursued both in AD and in models of AD as a potential means for treating AD. Both active and passive immunizations targeted at fibrillar Abeta successfully remove cerebral plaque load and attenuate Abeta-induced toxicity. Consistently with this, intracerebroventricular (ICV) passive immunization established in our laboratory using antibody against fibrillar Abeta (anti-fAbeta) reduced cerebral plaque load and reversed early synaptic deficits at pre/early plaque stage when there is an abundance of soluble dimeric/oligomeric Abeta but sparse fibrillar Abeta, indicating that anti-fAbeta-mediated partial neutralization of toxic oligomeric Abeta species might have reduced early synaptotoxicity. In the previous investigation, we found that immunoneutralization with anti-fAbeta transiently reduced cerebral Abeta and associated toxicity. The current investigation tested whether ICV im munization using antibody to conformationally changed oligomeric Abeta (anti-oligoAbeta) will overcome the transient restorative nature of anti-fAbeta and produce persistent, long-lasting preventive effects. Because oligomeric Abeta is strongly correlated with synaptotoxicity, we investigated whether immunoneutralization of oligomeric Abeta will reverse synaptic deficits by analyzing presynaptic molecular marker (SNAP-25) profile within hippocampal dendritic fields, where SNAP-25 is abundantly expressed. Results show that, in contrast to ICV anti-fAbeta antibody, ICV anti-oligoAbeta antibody significantly prevented cerebral Abeta build and almost completely restored SNAP-25 immunoreaction up to 8 weeks postinjection in TgCRND8 brain. Results show that ICV passive immunization with anti-oligoAbeta antibody might be an improved ICV immunization strategy for preventing permanent structural damage in AD.

Effect of aged garlic extract on APP processing and tau phosphorylation in Alzheimer's transgenic model Tg2576.

Multiple components present in garlic and various garlic preparations are known to exert pleiotropic protective effects as demonstrated in various in vitro and in vivo model systems. However, garlic pleiotropy in relation to Alzheimer's pathophysiology has not been explored extensively. Current study investigated anti-amyloidogenic, anti-inflammatory and anti-tangle effects of dietary aged garlic extract (AGE) (2%) and compared with its prominent constituents, i.e. S-allyl-cysteine (SAC) (20 mg/kg) and di-allyl-disulfide (DADS) (20 mg/kg) in Alzheimer's Swedish double mutant mouse model (Tg2576). Possible cholesterol-dependent and cholesterol-independent mechanisms of actions of AGE, SAC and DADS in exerting anti-amyloidogenic, anti-inflammatory and anti-tangle effects are discussed. Finally, ameliorative effects of dietary interventions were found to be in the order of AGE>SAC>DADS. If validated pre-clinically, dietary intervention with herbal alternative such as AGE having pleiotropic useful properties and least adverse effects may provide greater therapeutic benefit over a single-ingredient synthetic pharmaceutical drug having serious side effects in treating Alzheimer's disease.

Beta-amyloid-dependent expression of NOS2 in neurons: prevention by an alpha2-adrenergic antagonist.

The neurotransmitter noradrenaline (NA) exerts important antiinflammatory effects on glial cells including suppression of the inducible form of nitric oxide synthase (NOS2). The authors examined the consequences of manipulating NA in vivo by treating adult rats with the neurotoxin DSP4, which selectively lesions noradrenergic neurons of the locus ceruleus (LC), and reduces cortical NA levels. Following LC lesion, intracortical injection of aggregated amyloid beta 1-42 (Abeta1-42) caused appearance of NOS2 within neurons, and increased neuronal damage assessed by staining for nonphosphorylated neurofilament proteins with antibody SMI-32. Co-treatment with a selective alpha2-adrenergic antagonist reduced neuronal NOS2 staining as well as SMI-32 staining. Neuronal damage was dependent on NOS2 expression since injection of Abeta1-42 into DSP4-treated NOS2-deficient mice did not result in neuronal damage. These results demonstrate that decrease of NA levels in vivo can exacerbate inflammatory responses and neuronal damage due to inflammatory stimuli such as Abeta. These findings suggest that alpha2-adrenergic antagonists could provide therapeutic benefit in neurological diseases such as AD or PD where LC loss is known to occur.

Efficacy of anti-Abeta antibody isotypes used for intracerebroventricular immunization in TgCRND8.

We have previously demonstrated that intracerebroventricular (ICV) injection of anti-Abeta (IgG1, kappa against the 1-28 region of Abeta) reduced cerebral amyloid plaques by 50% after 1 month without producing hemorrhage or activating IL-1beta responses in Tg2576 brain [N.B. Chauhan, G.J. Siegel, Reversal of amyloid beta toxicity in Alzheimer's disease model Tg2576 by intraventricular antiamyloid beta antibody, J. Neurosci. Res. 69 (1) (2002) 10-23]. The current report compares the efficacy of IgG1, IgG2a and IgG2b isotypes of anti-Abeta against several different epitopes of Abeta in clearing cerebral Abeta after a single bolus ICV injection in TgCRND8. Consistent with earlier in vitro findings from other laboratories, these in vivo data demonstrate that all IgG1 isotype antibodies tested cleared cerebral Abeta more efficiently than did IgG2a and IgG2b antibodies without producing histotoxicity in brain, liver or kidney, while an antibody against the C-terminus of Abeta did not reduce plaques or diminish their accumulation with aging of the animals. Intriguingly, there was no significant difference between the Abeta-reducing efficiency of IgG1 anti-Abeta antibodies directed against residues 3-6, against residues 1-10 or against residues 1-28 of N-terminus Abeta.

Propentofylline attenuates tau hyperphosphorylation in Alzheimer's Swedish mutant model Tg2576.

Key pathological hallmarks of Alzheimer's disease (AD) are the deposition of amyloid plaques containing Abeta-peptides and the formation of neurofibrillary tangles containing hyperphosphorylated tau. Propentofylline (PPF) is a synthetic xanthine derivative that inhibits phosphodiesterase and adenosine uptake. These effects of PPF influence many cellular functions including stimulating synthesis/release of nerve growth factor. We tested the effects of PPF on disease progression in transgenic mice overexpressing the Swedish mutant human APP (Tg2576). The untreated Tg mice show, together with increased amyloidogenesis, increased levels of tau hyperphosphorylation and increased ratios of the activated to inactivated GSK-3beta, one of the key kinases that can phosphorylate tau. One month of PPF feeding (40 mg/kg per day) reduced the burden of amyloid plaques and the levels of hyperphosphorylated tau and immunoreactive IL-1beta. In parallel with these changes, PPF reduced the activated form of GSK-3beta and increased the inactivated form of GSK-3beta, restoring their ratio almost to normal values. These results demonstrate that PPF can exert multiple protective effects on both amyloidogenesis and tau hyperphosphorylation in an animal model of AD. Our earlier report [Neurochem. Int. 43(3) (2003) 225] demonstrated that Tg2576 animals show decreased levels of mRNA for NGF with increased amyloid burden while feeding of PPF results in a major shift from beta-amyloidogenic to alpha-secretory processing of APP together with increased expression of NGF mRNA. The current new data enlarge our understanding of PPF effects in brain and of tau hyperphosphorylation in Tg animals and are consistent with the hypothesis that GSK-3beta is a nodal point linking amyloid and tau pathology. Therapeutic interventions directed toward multiple pathological processes may be more protective than treatments directed toward a single process. The new results reported here indicate that further testing of PPF as a potential therapy in AD is warranted.