Brain Aggregates Research Articles

Systemic inflammation activates coagulation and immune cell infiltration pathways in brains with propagating α-synuclein fibril aggregates

Synucleinopathies are a group of diseases characterized by brain aggregates of α-synuclein (α-syn). The gradual accumulation of α-syn and the role of inflammation in early-stage pathogenesis remain poorly understood. We explored this interaction by inducing chronic inflammation in a common pre-clinical synucleinopathy mouse model. Three weeks post unilateral intra-striatal injections of human α-syn pre-formed fibrils (PFF), mice underwent repeated intraperitoneal injections of 1 mg/ml lipopolysaccharide (LPS) for 3 weeks. Histological examinations of the ipsilateral site showed phospho-α-syn regional spread and LPS-induced neutrophil recruitment to the brain vasculature. Biochemical assessment of the contralateral site confirmed spreading of α-syn aggregation to frontal cortex and a rise in intracerebral TNF-α, IL-1β, IL-10 and KC/GRO cytokines levels due to LPS. No LPS-induced exacerbation of α-syn pathology load was observed at this stage. Proteomic analysis was performed contralateral to the PFF injection site using LC-MS/MS. Subsequent downstream Reactome Gene-Set Analysis indicated that α-syn pathology alters mitochondrial metabolism and synaptic signaling. Chronic LPS-induced inflammation further lead to an overrepresentation of pathways related to fibrin clotting as well as integrin and B cell receptor signaling. Western blotting confirmed a PFF-induced increase in fibrinogen brain levels and a PFF + LPS increase in Iba1 levels, indicating activated microglia. Splenocyte profiling revealed changes in T and B cells, monocytes, and neutrophils populations due to LPS treatment in PFF injected animals. In summary, early α-syn pathology impacts energy homeostasis pathways, synaptic signaling and brain fibrinogen levels. Concurrent mild systemic inflammation may prime brain immune pathways in interaction with peripheral immunity.

CSF proteome profiling identifies novel biomarkers for Frontotemporal Dementia and its pathological subtypes

AbstractBackgroundFrontotemporal dementia (FTD) is caused by frontotemporal lobar degeneration (FTLD) and the most common forms are characterized by either tau (FTLD‐Tau) or TDP43 (FTLD‐TDP) brain aggregates. However, FTD‐specific fluid biomarkers are lacking. Furthermore, the pathological subtypes are not distinct in their presentation, hampering accurate subtyping at clinical diagnosis. Therefore, there is a strong need to identify fluid biomarkers that could aid in FTD diagnosis and to discriminate the pathological subtypes.MethodWe employed an antibody‐based proteomic technology to analyze >600 proteins in a large multicenter cohort including cerebrospinal fluid (CSF) samples from FTD (n = 189), AD (n = 235) and cognitively unimpaired individuals (n = 196). For a subset of cases the underlying neuropathology was known or could be predicted (FTLD‐Tau = 85 and FTLD‐TDP = 57). Differences in protein expression profiles were analyzed by nested linear models. Penalized generalized linear modeling was used to identify classification protein panels. Protein panels were then validated in independent clinical cohorts (cohort 1: n = 157; cohort 2: n = 165) and a neuropathology cohort (n = 100) using customized assays.ResultWe observed 65 differentially regulated proteins in FTD versus controls and AD patients, associated with axonogenesis, synapse assembly, or locomotory behavior pathways. We identified panels of 14 and 13 proteins that could discriminate FTD from controls (AUC = 0.96, 95%CI:0.91‐0.99) and AD patients (AUC = 0.91, 95%CI:0.85‐0.96), respectively. Most of these proteins (21 out of 27) were translated into customized panels, which discriminated between groups with high accuracy for all three cohorts (FTDvsCon: AUCs > 0.96, FTDvsAD: AUCs > 0.88). When comparing the FTLD‐Tau and FTLD‐TDP subtypes, we observed that 86 proteins were increased in FTLD‐Tau, and associated with developmental and cellular processes and locomotion pathways. A panel of 8 proteins could discriminate between the pathological subtypes (AUC = 0.82, 95%CI:0.66‐0.95), which was however not stable during cross‐validation.ConclusionWe identified and validated CSF panels to discriminate FTD from controls and AD with high accuracy. The identification of a biomarker panel to discriminate between the FTLD pathological subtypes likely requires larger and more homogeneous groups. The panels developed within this study might be useful for diagnosis and trial inclusion of FTD patients.

Characteristics of AD‐brain tau aggregate internalisation by human astrocytes and the responses induced

AbstractBackgroundHighly phosphorylated tau aggregates are deposited in tauopathy brain as disease progresses, driven by a prion‐like spread of tau seeds. Evidence suggests that astrocytes may influence tau spread. However, the efficiency of uptake of disease‐associated tau species is not well defined, nor are the effects of tau uptake on astrocyte reactivity and function. We investigate human astrocyte ability to internalise and clear tau aggregates derived from multiple Alzheimer’s disease (AD) cases, and their impact on astrocyte function and reactivity. We further explore alterations in endogenous tau expression, APP processing and cytoskeletal proteins following tau uptake.MethodControl human iPSC‐derived astrocytes were cultured for 70 days. Aggregated (sarkosyl‐insoluble) tau was isolated from postmortem AD brain. Post‐translational modifications (PTM) of isolated AD‐brain tau aggregates were analysed by mass spectrometry. Tau (0.1 ng/µL) and extracts from control human brain were spiked into astrocyte cultures. Internalisation of aggregated tau and the localised response of S100B and GFAP were quantified by ICC. Changes to tau, APP, functional and reactivity related genes were measured by qPCR and/or proteins by SDS‐PAGE.ResultAggregated tau from AD brain is readily internalised by human astrocytes, and sequesters GFAP and S100B. Differences in the rate of uptake, clearance and seeding of tau correlates with cytoskeletal protein accumulation and specific changes in astrocyte mRNA and protein levels. Work is in progress to relate these changes to characteristics of tau aggregates.ConclusionOur data shows that the capacity of AD brain aggregates to be internalised, cleared and induce seeding of endogenous tau varies by case. Investigations are underway to pinpoint the tau characteristics that underlie these differences. We also show that seeding ability is associated with changes in cytoskeletal elements and reactivity markers at mRNA and protein level. This work agrees with growing evidence that tau variability in AD is linked with specific cell type vulnerabilities that contribute to tau spread and disease progression.

USP19 deubiquitinase inactivation regulates α-synuclein ubiquitination and inhibits accumulation of Lewy body-like aggregates in mice

The USP19 deubiquitinase is found in a locus associated with Parkinson’s Disease (PD), interacts with chaperonins, and promotes secretion of α-synuclein (α-syn) through the misfolding-associated protein secretion (MAPS) pathway. Since these processes might modulate the processing of α-syn aggregates in PD, we inactivated USP19 (KO) in mice expressing the A53T mutation of α-syn and in whom α-syn preformed fibrils (PFF) had been injected in the striatum. Compared to WT, KO brains showed decreased accumulation of phospho-synuclein (pSyn) positive aggregates. This improvement was associated with less activation of microglia and improved performance in a tail-suspension test. Exposure of primary neurons from WT and KO mice to PFF in vitro also led to decreased accumulation of pSyn aggregates. KO did not affect uptake of PFF nor propagation of aggregates in the cultured neurons. We conclude that USP19 instead modulates intracellular dynamics of aggregates. At an early time following PFF injection when the number of pSyn-positive neurons were similar in WT and KO brains, the KO neurons contained less aggregates. KO brain aggregates stained more intensely with anti-ubiquitin antibodies. Immunoprecipitation of soluble proteins from WT and KO brains with antibodies to pSyn showed higher levels of ubiquitinated oligomeric species in the KO samples. We propose that the improved pathology in USP19 KO brains may arise from decreased formation or enhanced clearance of the more ubiquitinated aggregates and/or enhanced disassembly towards more soluble oligomeric species. USP19 inhibition may represent a novel therapeutic approach that targets the intracellular dynamics of α-syn complexes.

Neural representation of gestalt grouping and attention effect in human visual cortex

BackgroundThe brain aggregates meaningless local sensory elements to form meaningful global patterns in a process called perceptual grouping. Current brain imaging studies have found that neural activities in V1 are modulated during visual grouping. However, how grouping is represented in each of the early visual areas, and how attention alters these representations, is still unknown. New methodWe adopted MVPA to decode the specific content of perceptual grouping by comparing neural activity patterns between gratings and dot lattice stimuli which can be grouped with proximity law. Furthermore, we quantified the grouping effect by defining the strength of grouping, and assessed the effect of attention on grouping. ResultsWe found that activity patterns to proximity grouped stimuli in early visual areas resemble these to grating stimuli with the same orientations. This similarity exists even when there is no attention focused on the stimuli. The results also showed a progressive increase of representational strength of grouping from V1 to V3, and attention modulation to grouping is only significant in V3 among all the visual areas. Comparison with existing methodsMost of the previous work on perceptual grouping has focused on how activity amplitudes are modulated by grouping. Using MVPA, the present work successfully decoded the contents of neural activity patterns corresponding to proximity grouping stimuli, thus shed light on the availability of content-decoding approach in the research on perceptual grouping. ConclusionsOur work found that the content of the neural activity patterns during perceptual grouping can be decoded in the early visual areas under both attended and unattended task, and provide novel evidence that there is a cascade processing for proximity grouping through V1 to V3. The strength of grouping was larger in V3 than in any other visual areas, and the attention modulation to the strength of grouping was only significant in V3 among all the visual areas, implying that V3 plays an important role in proximity grouping.

Specific post-translational modifications of soluble tau protein distinguishes Alzheimer’s disease and primary tauopathies

Tau protein aggregates in several neurodegenerative disorders, referred to as tauopathies. The tau isoforms observed in post mortem human brain aggregates is used to classify tauopathies. However, distinguishing tauopathies ante mortem remains challenging, potentially due to differences between insoluble tau in aggregates and soluble tau in body fluids. Here, we demonstrated that tau isoforms differ between tauopathies in insoluble aggregates, but not in soluble brain extracts. We therefore characterized post-translational modifications of both the aggregated and the soluble tau protein obtained from post mortem human brain tissue of patients with Alzheimer’s disease, cortico-basal degeneration, Pick’s disease, and frontotemporal lobe degeneration. We found specific soluble signatures for each tauopathy and its specific aggregated tau isoforms: including ubiquitination on Lysine 369 for cortico-basal degeneration and acetylation on Lysine 311 for Pick’s disease. These findings provide potential targets for future development of fluid-based biomarker assays able to distinguish tauopathies in vivo.

Tau mutation S356T in the three repeat isoform leads to microtubule dysfunction and promotes prion-like seeded aggregation.

Tauopathies are a group of neurodegenerative diseases, which include frontotemporal dementia (FTD) and Alzheimer's disease (AD), broadly defined by the development of tau brain aggregates. Both missense and splicing tau mutations can directly cause early onset FTD. Tau protein is a microtubule-associated protein that stabilizes and regulates microtubules, but this function can be disrupted in disease states. One contributing factor is the balance of different tau isoforms, which can be categorized into either three repeat (3R) or four repeat (4R) isoforms based on the number of microtubule-binding repeats that are expressed. Imbalance of 3R and 4R isoforms in either direction can cause FTD and neurodegeneration. There is also increasing evidence that 3R tauopathies such as Pick's disease form tau aggregates predominantly comprised of 3R isoforms and these can present differently from 4R and mixed 3R/4R tauopathies. In this study, multiple mutations in 3R tau were assessed for MT binding properties and prion-like aggregation propensity. Different missense tau mutations showed varying effects on MT binding depending on molecular location and properties. Of the mutations that were surveyed, S356T tau is uniquely capable of prion-like seeded aggregation and forms extensive Thioflavin positive aggregates. This unique prion-like tau strain will be useful to model 3R tau aggregation and will contribute to the understanding of diverse presentations of different tauopathies.

TAT‐LBD‐Ngn2‐improved cognitive functions after global cerebral ischemia by enhancing neurogenesis

Stroke is the major cause of adult neurocognitive disorders (NCDs), and presents a significant burden on both of the families and society. To improve the cerebral injury, we generated a blood-brain barrier penetrating peptide TAT-LBD-Ngn2, in which Ngn2 (Neurogenin2) is a classical preneural gene that enhances neurogenesis, and neural precursor cells survival and differentiation. We previously demonstrated that it has a short-term protective effect against cerebral ischemia-reperfusion injury.However, it is uncertain if TAT-LBD-Ngn2 could promote neurogenesis to exhibit long-term therapeutic impact. In present study, TAT-LBD-Ngn2 was administered for 14 or 28 days following bilateral common carotid arteries occlusion (BCCAO). After confirming that TAT-LBD-Ngn2 could cross the brain blood barrier and aggregate in the hippocampus, we conducted open field test, Morris water maze and contextual fear conditioning to examine the long-term effect of TAT-LBD-Ngn2 on cognition. We discovered that TAT-LBD-Ngn2 significantly improved the spatial and contextual learning and memory on both days 14 and 28 after BCCAO, while TAT-LBD-Ngn2 exhibited anxiolytic effect only on day 14, but had no effect on locomotion. Using western blot and immunofluorescence, TAT-LBD-Ngn2 was also shown to promote neurogenesis, as evidenced by increased BrdU+ and DCX+ neurons in dentate gyrus. Meanwhile, TAT-LBD-Ngn2 elevated the expression of brain derived neurotrophic factor rather than nerve growth factor compared to the control group. Our findings revealed that TAT-LBD-Ngn2 could dramatically promote learning and memory in long term by facilitating neurogenesis in the hippocampus after global cerebral ischemia, indicating that TAT-LBD-Ngn2 may be an appealing candidate for treating poststroke NCD.

Physiological Consequences of Targeting 14-3-3 and Its Interacting Partners in Neurodegenerative Diseases.

The mammalian 14-3-3 family comprises seven intrinsically unstructured, evolutionarily conserved proteins that bind >200 protein targets, thereby modulating cell-signaling pathways. The presence of 14-3-3 proteins in cerebrospinal fluid provides a sensitive and specific biomarker of neuronal damage associated with Alzheimer’s disease (AD), Creutzfeldt−Jakob disease (CJD), spongiform encephalitis, brain cancers, and stroke. We observed significant enrichment of 14-3-3 paralogs G, S, and Z in human brain aggregates diagnostic of AD. We used intra-aggregate crosslinking to identify 14-3-3 interaction partners, all of which were significantly enriched in AD brain aggregates relative to controls. We screened FDA-approved drugs in silico for structures that could target the 14-3-3G/hexokinase interface, an interaction specific to aggregates and AD. C. elegans possesses only two 14-3-3 orthologs, which bind diverse proteins including DAF-16 (a FOXO transcription factor) and SIR-2.1 (a sensor of nutrients and stress), influencing lifespan. Top drug candidates were tested in C. elegans models of neurodegeneration-associated aggregation and in a human neuroblastoma cell-culture model of AD-like amyloidosis. Several drugs opposed aggregation in all models assessed and rescued behavioral deficits in C. elegans AD-like neuropathy models, suggesting that 14-3-3 proteins are instrumental in aggregate accrual and supporting the advancement of drugs targeting 14-3-3 protein complexes with their partners.

Quantitative super-resolution imaging of pathological aggregates reveals distinct toxicity profiles in different synucleinopathies

Protein aggregation is a hallmark of major neurodegenerative disorders. Increasing data suggest that smaller aggregates cause higher toxic response than filamentous aggregates (fibrils). However, the size of small aggregates has challenged their detection within biologically relevant environments. Here, we report approaches to quantitatively super-resolve aggregates in live cells and ex vivo brain tissues. We show that Amytracker 630 (AT630), a commercial aggregate-activated fluorophore, has outstanding photophysical properties that enable super-resolution imaging of α-synuclein, tau, and amyloid-β aggregates, achieving ∼4 nm precision. Applying AT630 to AppNL-G-F mouse brain tissues or aggregates extracted from a Parkinson's disease donor, we demonstrate excellent agreement with antibodies specific for amyloid-β or α-synuclein, respectively, confirming the specificity of AT630. Subsequently, we use AT630 to reveal a linear relationship between α-synuclein aggregate size and cellular toxicity and discovered that aggregates smaller than 450 ± 60 nm (aggregate450nm) readily penetrated the plasma membrane. We determine aggregate450nm concentrations in six Parkinson's disease and dementia with Lewy bodies donor samples and show that aggregates in different synucleinopathies demonstrate distinct potency in toxicity. We further show that cell-penetrating aggregates are surrounded by proteasomes, which assemble into foci to gradually process aggregates. Our results suggest that the plasma membrane effectively filters out fibrils but is vulnerable to penetration by aggregates of 450 ± 60 nm. Together, our findings present an exciting strategy to determine specificity of aggregate toxicity within heterogeneous samples. Our approach to quantitatively measure these toxic aggregates in biological environments opens possibilities to molecular examinations of disease mechanisms under physiological conditions.

Learning to Feel Textures: Predicting Perceptual Similarities From Unconstrained Finger-Surface Interactions.

Whenever we touch a surface with our fingers, we perceive distinct tactile properties that are based on the underlying dynamics of the interaction. However, little is known about how the brain aggregates the sensory information from these dynamics to form abstract representations of textures. Earlier studies in surface perception all used general surface descriptors measured in controlled conditions instead of considering the unique dynamics of specific interactions, reducing the comprehensiveness and interpretability of the results. Here, we present an interpretable modeling method that predicts the perceptual similarity of surfaces by comparing probability distributions of features calculated from short time windows of specific physical signals (finger motion, contact force, fingernail acceleration) elicited during unconstrained finger-surface interactions. The results show that our method can predict the similarity judgments of individual participants with a maximum Spearman's correlation of 0.7. Furthermore, we found evidence that different participants weight interaction features differently when judging surface similarity. Our findings provide new perspectives on human texture perception during active touch, and our approach could benefit haptic surface assessment, robotic tactile perception, and haptic rendering.

Phosphorylated tau as a toxic agent in synaptic mitochondria: implications in aging and Alzheimer's disease.

During normal aging, there is a decline in all physiological functions in the organism. One of the most affected organs is the brain, where neurons lose their proper synaptic function leading to cognitive impairment. Aging is one of the main risk factors for the development of neurodegenerative diseases, such as Alzheimer’s disease. One of the main responsible factors for synaptic dysfunction in aging and neurodegenerative diseases is the accumulation of abnormal proteins forming aggregates. The most studied brain aggregates are the senile plaques, formed by Aβ peptide; however, the aggregates formed by phosphorylated tau protein have gained relevance in the last years by their toxicity. It is reported that neurons undergo severe mitochondrial dysfunction with age, with a decrease in adenosine 5′-triphosphate production, loss of the mitochondrial membrane potential, redox imbalance, impaired mitophagy, and loss of calcium buffer capacity. Interestingly, abnormal tau protein interacts with several mitochondrial proteins, suggesting that it could induce mitochondrial dysfunction. Nevertheless, whether tau-mediated mitochondrial dysfunction occurs indirectly or directly is still unknown. A recent study of our laboratory shows that phosphorylated tau at Ser396/404 (known as PHF-1), an epitope commonly related to pathology, accumulates inside mitochondria during normal aging. This accumulation occurs preferentially in synaptic mitochondria, which suggests that it may contribute to the synaptic failure and cognitive impairment seen in aged individuals. Here, we review the main tau modifications promoting mitochondrial dysfunction, and the possible mechanism involved. Also, we discuss the evidence that supports the possibility that phosphorylated tau accumulation in synaptic mitochondria promotes synaptic and cognitive impairment in aging. Finally, we show evidence and argue about the presence of phosphorylated tau PHF-1 inside mitochondria in Alzheimer’s disease, which could be considered as an early event in the neurodegenerative process. Thus, phosphorylated tau PHF-1 inside the mitochondria could be considered such a potential therapeutic target to prevent or attenuate age-related cognitive impairment.

Role of PLCγ1 in the modulation of cell migration and cell invasion in glioblastoma.

Phosphoinositide-specific phospholipases C (PLCs) are a class of enzymes involved in several cell activities, such as cell cycle regulation, proliferation, differentiation and cytoskeletal dynamics. Among these enzymes, PLCγ1 is one of the most expressed PLCs in the brain, contributing to a complex network in the developing nervous system. Several studies have shown that PLCγ1 signaling imbalance is linked to several brain disorders, including glioblastoma, the most aggressive brain tumor in adults. Indeed, it has been demonstrated a link between PLCγ1 inhibition and the arrest of glioma cell motility of fetal rat brain aggregates and the impairment of cell invasion abilities following its down-regulation. This study aims to determine the pathological influence of PLCγ1 in glioblastoma, through a translational study which combines in silico data, data from glioblastoma patients' samples and data on engineered cell lines. We found out that PLCγ1 gene expression correlates with the pathological grade of gliomas, and it is higher in fifty patients' glioblastoma tissue samples compared to twenty healthy controls. Moreover, it was demonstrated that PLCγ1 silencing in U87-MG leads to a reduction in cell migration and invasion abilities. The opposite trend was observed following PLCγ1 overexpression, suggesting an interesting possible involvement of PLCγ1 in gliomas' aggressiveness.

Disclosing tau tangles using PET imaging: a pharmacological review of the radiotracers available in 2021.

Neurological symptoms depend on the topography of the lesions in the nervous system, hence the importance of brain imaging for neurologists. Neurological treatment, however, depends on the biological nature of the lesions. The development of radiotracers specific for the proteinopathies observed in neurodegenerative disorders is, therefore, crucially important for better understanding the relationships between the pathology and the clinical symptoms, as well as the efficacy of therapeutical interventions. The tau protein is involved in several neurodegenerative disorders, that can be distinguished both biologically and clinically as the type of tau isoforms and filaments observed in brain aggregates, and the brain regions affected differ between tauopathies. Over the past few years, several tracers have been developed for imaging tauopathies with positron emission tomography. The present review aims to compare the binding properties of these tracers, with a specific focus on how these properties might be relevant for neurologists using these biomarkers to characterize the pathology of patients presenting with clinical symptoms suspect of a neurodegenerative disorder.

Analysis of circulating protein aggregates as a route of investigation into neurodegenerative disorders.

Plasma proteome composition reflects the inflammatory and metabolic state of the organism and can be predictive of system-level and organ-specific pathologies. Circulating protein aggregates are enriched with neurofilament heavy chain—axonal proteins involved in brain aggregate formation and recently identified as biomarkers of the fatal neuromuscular disorder amyotrophic lateral sclerosis. Using unbiased proteomic methods, we have fully characterized the content in neuronal proteins of circulating protein aggregates from amyotrophic lateral sclerosis patients and healthy controls, with reference to brain protein aggregate composition. We also investigated circulating protein aggregate protein aggregation propensity, stability to proteolytic digestion and toxicity for neuronal and endothelial cell lines. Circulating protein aggregates separated by ultracentrifugation are visible as electron-dense macromolecular particles appearing as either large globular or as small filamentous formations. Analysis by mass spectrometry revealed that circulating protein aggregates obtained from patients are enriched with proteins involved in the proteasome system, possibly reflecting the underlying basis of dysregulated proteostasis seen in the disease, while those from healthy controls show enrichment of proteins involved in metabolism. Compared to the whole human proteome, proteins within circulating protein aggregates and brain aggregates show distinct chemical features of aggregation propensity, which appear dependent on the tissue or fluid of origin and not on the health status. Neurofilaments’ two high-mass isoforms (460 and 268 kDa) showed a strong differential expression in amyotrophic lateral sclerosis compared to healthy control circulating protein aggregates, while aggregated neurofilament heavy chain was also partially resistant to enterokinase proteolysis in patients, demonstrated by immunoreactive bands at 171 and 31 kDa fragments not seen in digested healthy controls samples. Unbiased proteomics revealed that a total of 4973 proteins were commonly detected in circulating protein aggregates and brain, including 24 expressed from genes associated with amyotrophic lateral sclerosis. Interestingly, 285 circulating protein aggregate proteins (5.7%) were regulated (P < 0.05) and are present in biochemical pathways linked to disease pathogenesis and protein aggregation. Biologically, circulating protein aggregates from both patients and healthy controls had a more pronounced effect on the viability of hCMEC/D3 endothelial and PC12 neuronal cells compared to immunoglobulins extracted from the same plasma samples. Furthermore, circulating protein aggregates from patients exerted a more toxic effect than healthy control circulating protein aggregates on both cell lines at lower concentrations (P: 0.03, in both cases). This study demonstrates that circulating protein aggregates are significantly enriched with brain proteins which are representative of amyotrophic lateral sclerosis pathology and a potential source of biomarkers and therapeutic targets for this incurable disorder.

Targeted Quantification of Detergent-Insoluble RNA-Binding Proteins in Human Brain Reveals Stage and Disease Specific Co-aggregation in Alzheimer's Disease.

Core spliceosome and related RNA-binding proteins aggregate in Alzheimer’s disease (AD) brain even in early asymptomatic stages (AsymAD) of disease. To assess the specificity of RNA-binding protein aggregation in AD, we developed a targeted mass spectrometry approach to quantify broad classes of RNA-binding proteins with other pathological proteins including tau and amyloid beta (Aβ) in detergent insoluble fractions from control, AsymAD, AD and Parkinson’s disease (PD) brain. Relative levels of specific insoluble RNA-binding proteins across different disease groups correlated with accumulation of Aβ and tau aggregates. RNA-binding proteins, including splicing factors with homology to the basic-acidic dipeptide repeats of U1-70K, preferentially aggregated in AsymAD and AD. In contrast, PD brain aggregates were relatively depleted of many RNA-binding proteins compared to AsymAD and AD groups. Correlation network analyses resolved 29 distinct modules of co-aggregating proteins including modules linked to spliceosome assembly, nuclear speckles and RNA splicing. Modules related to spliceosome assembly and nuclear speckles showed stage-specific enrichment of insoluble RBPs from AsymAD and AD brains, whereas the RNA splicing module was reduced specifically in PD. Collectively, this work identifies classes of RNA-binding proteins that distinctly co-aggregate in detergent-insoluble fractions across the specific neurodegenerative diseases we examined.

Validity of tau as both a biomarker and target for AD and the dynamics of tau over the course of the disease

AbstractBackgroundBrain tau aggregates are, together with amyloid plaques, hallmarks of Alzheimer Disease (AD). Structural characterization of tau aggregates in AD, Pick’s disease and Chronic Traumatic Encephalopathy demonstrate distinct folds leading to tau aggregation. However, initiation and propagation mechanisms involved are still unknown. One hypothesis includes modification in tau metabolism affecting tau isoforms abundance and clearance. Understanding these mechanisms can both identify potential targets for treatment, biomarkers for monitoring disease stages and response to tau‐directed therapies.MethodRecent developments in characterizing tau species by Mass Spectrometry in brain and biofluids have provided new insights on the diversity of tau isoforms and their changes associated to Alzheimer’s disease over the course of the disease. We will review past and new results from these techniques and discuss which tau isoforms are the most promising biomarkers to identify and stage AD. We will then discuss how these changes can be related to abnormal tau metabolism and, potentially, aggregation.ResultIn brain, soluble tau is phosphorylated at more than 30 sites and tau phosphorylation occupancy is modified in AD in a site‐dependent manner and appears to occur decades before substantial tau aggregation. Contrarily, AD brain aggregates are hyperphosphorylated compared to AD soluble tau at specific sites. In cerebrospinal fluid (CSF), the longitudinal monitoring of 11 phosphorylated sites in autosomal dominant AD demonstrates different behavior of these sites over disease trajectory. The CSF tau truncation profile, particularly in the protein domain enriched in brain aggregates, changes with disease. This suggests brain tau aggregates may impact the nature of tau isoforms released extracellularly. Yet, initial stages of tau hyperphosphorylation appears to occur many years before substantial tau aggregation. Critically, certain tau hyperphosphorylation sites observed in CSF are recapitulated in plasma tau in the preclinical stages of disease.ConclusionBoth CSF tau phosphorylation and tau truncation profiles can serve as biomarkers to complement p‐tau and tau levels for AD diagnosis and staging. For target validation, the multiplicity of tau isoforms complicates the identification of the most relevant species to target. Notably, the uncertain role of abnormal tau phosphorylation as the initiator or a consequence of tau aggregation remains to be addressed.

Paradoxical cognitive reserve: Cognitive trajectories from earlier to later adulthood

AbstractBackgroundLongitudinal studies of aging are central to identifying early predictors of and mechanisms underlying late‐life cognitive decline and dementia. However, these studies typically lack cognitive data from earlier ages. It is therefore unclear how peak young adult general cognitive ability (GCA) and maintenance of GCA from young adulthood impact subsequent cognitive trajectories. To further understand GCA change from young adulthood, we evaluated whether poor GCA maintenance coincided with accelerated brain aging and was associated with key contributors to cognitive decline in later life, namely, aggregate vascular risk and Alzheimer’s disease polygenic risk (AD‐PRS).MethodIn the Vietnam Era Twin Study of Aging (VETSA), 1173 individuals were administered the same measure of GCA at average age 20 and at three VETSA waves beginning at average age 56. GCA maintenance scores were created by residualizing VETSA baseline GCA on age 20 GCA. Thus, good GCA maintenance was based on observed relative to expected scores. VETSA baseline GCA and GCA maintenance scores were entered into mixed effects models to predict specific cognitive abilities across three VETSA waves (∼6 years apart). Associations between GCA maintenance scores and indices of accelerated brain aging, aggregate vascular risk, and AD‐PRS were evaluated.ResultBetter GCA maintenance was associated with better VETSA baseline memory, semantic fluency, general fluency, processing speed, executive function, and working memory. However, given equivalent age 56 GCA, those who declined in GCA from age 20 outperformed those with stable GCA in memory, verbal fluency, executive function, and working memory at age 56. Thus, despite poor cognitive maintenance, some element of high peak cognitive reserve in early adulthood still confers benefit. All specific cognitive abilities declined over time, but neither VETSA baseline GCA nor GCA maintenance predicted extent of decline. Poor GCA maintenance from young adulthood was associated with accelerated brain aging and aggregate vascular risk, but not AD‐PRS.ConclusionBoth peak young adult GCA and GCA maintenance are important contributors to late midlife cognitive ability. Poor maintenance of GCA from young adulthood being associated with worse vascular health and coinciding with accelerated brain aging argues against a simple regression to the mean explanation.

Allium roseum L. extract inhibits amyloid beta aggregation and toxicity involved in Alzheimer's disease.

Allium roseum is an important medicinal and aromatic plant, specific to the North African flora and a rich source of important nutrients and bioactive molecules including flavonoids and organosulfur compounds whose biological activities and pharmacological properties are well known. In the present study, the inhibition of amyloid beta protein toxicity by the ethanolic extract of this plant is investigated for the first time. Preliminary biochemical analyses identified kæmpferol and luteolin-7-o-glucoside as the more abundant phenolic compounds. The effects of A. roseum extract (ARE) on aggregation and aggregate cytotoxicity of amyloid beta-42 (Aβ42), whose brain aggregates are a hallmark of Alzheimer’s disease, were investigated by biophysical (ThT assay, Dynamic light scattering and transmission electron microscopy) and cellular assays (cytotoxicity, aggregate immunolocalization, ROS measurement and intracellular Ca2+ imaging). The biophysical data suggest that ARE affects the structure of the Aβ42 peptide, inhibits its polymerization, and interferes with the path of fibrillogenesis. The data with cultured cells shows that ARE reduces Aß42 aggregate toxicity by inhibiting aggregate binding to the cell membrane and by decreasing both oxidative stress and intracellular Ca2+. Accordingly, ARE could act as a neuroprotective factor against Aβ aggregate toxicity in Alzheimer’s disease.

Identification of amyloid plaques in the brain using an x-ray photon-counting strip detector.

Brain aggregates of β amyloid (βA) protein plaques have been widely recognized as associated with many neurodegenerative diseases, and their identification can assist in the early diagnosis of Alzheimer’s disease. We investigate the feasibility of using a spectral x-ray coherent scatter system with a silicon strip photon-counting detector for identifying brain βA protein plaques. This approach is based on differences in the structure of amyloid, white and grey matter in the brain. We simulated an energy- and angular-dispersive X-ray diffraction system with an x-ray pencil beam and Silicon strip sensor, energy-resolving detectors. The polychromatic beam is geometrically focused toward a region of interest in the brain. First, the open-source MC-GPU code for Monte Carlo transport was modified to accommodate the detector model. Second, brain phantoms with and without βA were simulated to assess the method and determine the radiation dose required to obtain acceptable statistical power. For βA targets of 3, 4 and 5 mm sizes in a 15-cm brain model, the required incident exposure was about 0.44 mR from a 60 kVp tungsten spectrum and 3.5 mm of added aluminum filtration. The results suggest that the proposed x-ray coherent scatter technique enables the use of high energy x-ray spectra and therefore has the potential to be used for accurate in vivo detection and quantification of βA in the brain within acceptable radiation dose levels.