Functional BRI2-TREM2 interactions in microglia: implications for Alzheimer's and related dementias.

ITM2B/BRI2 mutations cause Alzheimer's Disease (AD)-related dementias. We observe heightened ITM2B/BRI2 expression in microglia, a pivotal cell type in AD due to risk-increasing variants in the microglial gene TREM2. Single-cell RNA-sequencing demonstrates a Trem2/Bri2-dependent microglia cluster, underscoring their functional interaction. α-secretase cleaves TREM2 into TREM2-CTF and sTREM2. As BRI2 hinders α-secretase cleavage of the AD-related Aß-Precursor-Protein, we probed whether BRI2 influences TREM2 processing. Our findings indicate a BRI2-TREM2 interaction that inhibits TREM2 processing in heterologous cells. Recombinant BRI2 and TREM2 proteins demonstrate a direct, cell-free BRI2-TREM2 ectodomain interaction. Constitutive and microglial-specific Itm2b-Knock-out mice, and Itm2b-Knock-out primary microglia provide evidence that Bri2 reduces Trem2 processing, boosts Trem2 mRNA expression, and influences Trem2 protein levels through α-secretase-independent pathways, revealing a multifaceted BRI2-TREM2 functional interaction. Moreover, a mutant Itm2b dementia mouse model exhibits elevated Trem2-CTF and sTrem2, mirroring sTREM2 increases in AD patients. Lastly, Bri2 deletion reduces phagocytosis similarly to a pathogenic TREM2 variant that enhances processing. Given BRI2's role in regulating Aß-Precursor-Protein and TREM2 functions, it holds promise as a therapeutic target for AD and related dementias.

Aß43 levels determine the onset of pathological amyloid deposition.

About 2% of Alzheimer's disease (AD) cases have early onset (FAD) and are caused by mutations in either Presenilins (PSEN1/2) or amyloid-ß precursor protein (APP). PSEN1/2 catalyze production of Aß peptides of different length from APP. Aß peptides are the major components of amyloid plaques, a pathological lesion that characterizes AD. Analysis of mechanisms by which PSEN1/2 and APP mutations affect Aß peptide compositions lead to the implication of the absolute or relative increase in Aß42 in amyloid-ß plaques formation. Here, to elucidate the formation of pathogenic Aß cocktails leading to amyloid pathology, we utilized FAD rat knock-in models carrying the Swedish APP (Apps allele) and the PSEN1 L435F (Psen1LF allele) mutations. To accommodate the differences in the pathogenicity of rodent and human Aß, these rat models are genetically engineered to express human Aß species as both the Swedish mutant allele and the WT rat allele (called Apph) have been humanized in the Aß-coding region. Analysis of the eight possible FAD mutant permutations indicates that the CNS levels of Aß43, rather than absolute or relative increases in Aß42, determine the onset of pathological amyloid deposition in FAD knock-in rats. Notably, Aß43 was found in amyloid plaques in late onset AD and mild cognitive impairment cases, suggesting that the mechanisms initiating amyloid pathology in FAD knock-in rat reflect disease mechanisms driving amyloid pathology in late onset AD. This study helps clarifying the molecular determinants initiating amyloid pathology and supports therapeutic interventions targeting Aß43 in AD.

GeneSelectML: a comprehensive way of gene selection for RNA-Seq data via machine learning algorithms.

Selection of differentially expressed genes (DEGs) is a vital process to discover the causes of diseases. It has been shown that modelling of genomics data by considering relation among genes increases the predictive performance of methods compared to univariate analysis. However, there exist serious differences among most studies analyzing the same dataset for the reasons arising from the methods. Therefore, there is a strong need for easily accessible, user-friendly, and interactive tool to perform gene selection for RNA-seq data via machine learning algorithms simultaneously not to miss DEGs. We develop an open-source and freely available web-based tool for gene selection via machine learning algorithms that can deal with high performance computation. This tool includes six machine learning algorithms having different aspects. Moreover, the tool involves classical pre-processing steps; filtering, normalization, transformation, and univariate analysis. It also offers well-arranged graphical approaches; network plot, heatmap, venn diagram, and box-and-whisker plot. Gene ontology analysis is provided for both mRNA and miRNA DEGs. The implementation is carried out on Alzheimer RNA-seq data to demonstrate the use of this web-based tool. Eleven genes are suggested by at least two out of six methods. One of these genes, hsa-miR-148a-3p, might be considered as a new biomarker for Alzheimer's disease diagnosis. Kidney Chromophobe dataset is also analyzed to demonstrate the validity of GeneSelectML web tool on a different dataset. GeneSelectML is distinguished in that it simultaneously uses different machine learning algorithms for gene selection and can perform pre-processing, graphical representation, and gene ontology analyses on the same tool. This tool is freely available at www.softmed.hacettepe.edu.tr/GeneSelectML .

Late-long-term potentiation magnitude, but not Aß levels and amyloid pathology, is associated with behavioral performance in a rat knock-in model of Alzheimer disease.

Cleavage of Amyloid precursor protein by ß- and γ-secretases lead to Aß formation. The widely accepted pathogenic model states that these mutations cause AD via an increase in Aß formation and accumulation of Aß in Amyloid plaques. APP mutations cause early onset familial forms of Alzheimer's disease (FAD) in humans. We generated App-Swedish (Apps ) knock-in rats, which carry a pathogenic APP mutation in the endogenous rat App gene. This mutation increases ß-secretase processing of APP leading to both augmented Aß production and facilitation of glutamate release in Apps/s rats, via a ß-secretase and APP-dependent glutamate release mechanism. Here, we studied 11 to 14-month-old male and female Apps/s rats. To determine whether the Swedish App mutation leads to behavioral deficits, Apps/s knock-in rats were subjected to behavioral analysis using the IntelliCage platform, an automated behavioral testing system. This system allows behavioral assessment in socially housed animals reflecting a more natural, less stress-inducing environment and eliminates experimenter error and bias while increasing precision of measurements. Surprisingly, a spatial discrimination and flexibility task that can reveal deficits in higher order brain function showed that Apps/s females, but not Apps/s male rats, performed significantly worse than same sex controls. Moreover, female control rats performed significantly better than control and Apps/s male rats. The Swedish mutation causes a significant increase in Aß production in 14-month-old animals of both sexes. Yet, male and female Apps/s rats showed no evidence of AD-related amyloid pathology. Finally, Apps/s rats did not show signs of significant neuroinflammation. Given that the APP Swedish mutation causes alterations in glutamate release, we analyzed Long-term potentiation (LTP), a long-lasting form of synaptic plasticity that is a cellular basis for learning and memory. Strikingly, LTP was significantly increased in Apps/s control females compared to both Apps/s sexes and control males. In conclusion, this study shows that behavioral performances are sex and App-genotype dependent. In addition, they are associated with LTP values and not Aß or AD-related pathology. These data, and the failures of anti-Aß therapies in humans, suggest that alternative pathways, such as those leading to LTP dysfunction, should be targeted for disease-modifying AD therapy.

Effects of chronic vagal nerve stimulation in the treatment of ß-amyloid-induced neuropsychiatric symptoms.

Alzheimer's Disease (AD) is the leading cause of dementia and, at the time of diagnosis, half of AD patients display at least one neuropsychiatric symptom (NPS). However, there is no effective therapy for NPSs; furthermore, current treatments of NPSs accelerate cognitive decline. Due to the ineffectiveness and negative consequences of current treatments for NPSs, new approaches are strongly needed. Currently, indications for vagal nerve stimulation (VNS) include epilepsy, stroke rehabilitation and major depression but not NPSs or AD. Therefore, we investigated whether chronic VNS can treat NPSs in a rat model of AD. Here, we report the intracerebroventricular injection of amyloid-ß (Aß) results in depression-like behaviors and memory impairment in rats. Chronic VNS (0.8 mA, 500 µs, 30 Hz, 5 min/day) showed strong antidepressant and anxiolytic effects, and improved memory performance. Additionally, the anxiolytic effect of VNS was retained in the non-Aß-treated rats. VNS also decreased aggressiveness and increased locomotor activity in both Aß-treated and non-Aß-treated rats. Recent studies showed VNS alters glutamatergic receptor levels, thus levels of GluA1, GluN2A, and GluN2B were determined. A significant reduction in GluN2B levels was seen in the hippocampus of VNS-treated groups which may relate to the anxiolytic effects and increased locomotor activity of VNS. In conclusion, VNS could be an effective treatment of NPSs, especially depression and anxiety, in AD patients without impairing cognition.

Fabrication of a 3D Printed PCL Nerve Guide: In Vitro and In Vivo Testing.

Nerve guides are medical devices designed to guide proximal and distal ends of injured peripheral nerves in order to assist regeneration of the damaged nerves. A 3D-printed polycaprolactone (PCL) nerve guide using an aligned gelatin-poly(3-hydroxybutyrate-co-3-hydroxyvalerate) electrospun mat, seeded with PC12 and Schwann cells (SCs) is produced. During characterization with microCT and SEM porosity (55%), pore sizes (675 ± 40 µm), and fiber diameters (382 ± 25 µm) are determined. Electrospun fibers have degree of alignment of 7°, indicating high potential for guidance. On Day 14, PC12 cells migrated from proximal to distal end of nerve guide when SCs are seeded on the guide. After 28 days, over 95% of PC12 are alive and aligned. PC12 cells express early differentiation marker beta-tubulin 10 times more than late marker NeuN. In a 10 mm rat sciatic nerve injury, functional recovery evaluated by using static sciatic index (SSI) is observed in mat-free guides and guides containing mat and SCs. Nerve conduction velocities are also improved in these groups. Histological stainings showed tissue growth around nerve guides with highest new tissue organization being observed with mat and cell-free guides. These suggest 3D-printed PCL nerve guides have significant potential for treatment of peripheral nerve injuries.

Association of entirely claims-based frailty indices with long-term outcomes in patients with acute myocardial infarction, heart failure, or pneumonia: a nationwide cohort study in Turkey.

BACKGROUND: Several countries have increasingly focused on improving care for acute myocardial infarction (AMI), heart failure (HF), and pneumonia to reduce their readmissions and mortality rates. Frailty is becoming increasingly important to accurately predict healthcare utilization for the aging population. The preferred method for the measurement of frailty remains unclear, and current risk-adjustment models do not account for frailty. We sought to compare commonly used frailty indices in terms of the ability to predict clinical adverse outcomes in AMI, HF, and pneumonia patients. METHODS: A nationwide cohort study included AMI, HF, and pneumonia with 65 years and older patients in the Turkey between January 1 and December 31, 2018. The primary predictor of interest was frailty. We used two claims-based frailty indices (Johns Hopkins Claims-Based Frailty Index and Hospital Frailty Risk Score) to assess frailty. The main outcome was all-cause long-term mortality up to 3 years. Time to death was calculated as the time period between the date of first admission and the date of death. Patients were censored as of September 30, 2020, which marked the end of the follow-up period. FINDINGS: Of the 200,948 patients, 35,096 (17.5%) had AMI, 62,403 (31.1%) had HF, and 103,449 (51.5%) had pneumonia. Johns Hopkins Claims-Based Frailty Index (c-statistics for long-term mortality: 0.68 in AMI, 0.61 in HF, 0.64 in pneumonia) was better compared to Hospital Frailty Risk Score (c-statistics for long-term mortality: AMI=0.62, HF=0.58, pneumonia=0.62) (DeLong p<0.001 in all). INTERPRETATION: Readmission and mortality rates after AMI, HF, and pneumonia gradually increases with increasing frailty score. While the Hospital Frailty Risk Score had a better discrimination for predicting readmissions, Johns Hopkins Claims-Based Frailty Index had a better discrimination for predicting mortality. These findings should be taken into account for a better evaluation of hospital performance. FUNDING: This study was supported by funding from The Scientific and Technological Research Council of Turkey (grant 120S422, HK).

High-fructose corn syrup consumption in adolescent rats causes bipolar-like behavioural phenotype with hyperexcitability in hippocampal CA3-CA1 synapses.

BACKGROUND AND PURPOSE: Children and adolescents are the top consumers of high-fructose corn syrup (HFCS) sweetened beverages. Even though the cardiometabolic consequences of HFCS consumption in adolescents are well known, the neuropsychiatric consequences have yet to be determined. EXPERIMENTAL APPROACH: Adolescent rats were fed for a month with 11% weight/volume carbohydrate containing HFCS solution, which is similar to the sugar-sweetened beverages of human consumption. The metabolic, behavioural and electrophysiological characteristics of HFCS-fed rats were determined. Furthermore, the effects of TDZD-8, a highly specific GSK-3B inhibitor, on the HFCS-induced alterations were further explored. KEY RESULTS: HFCS-fed adolescent rats displayed bipolar-like behavioural phenotype with hyperexcitability in hippocampal CA3-CA1 synapses. This hyperexcitability was associated with increased presynaptic release probability and increased readily available pool of AMPA receptors to be incorporated into the postsynaptic membrane, due to decreased expression of the neuron-specific α3-subunit of Na+ /K+ -ATPase and an increased ser845 -phosphorylation of GluA1 subunits (AMPA receptor subunit) respectively. TDZD-8 treatment was found to restore behavioural and electrophysiological disturbances associated with HFCS consumption by inhibition of GSK-3B, the most probable mechanism of action of lithium for its mood-stabilizing effects. CONCLUSION AND IMPLICATIONS: This study shows that HFCS consumption in adolescent rats led to a bipolar-like behavioural phenotype with neuronal hyperexcitability, which is known to be one of the earliest endophenotypic manifestations of bipolar disorder. Inhibition of GSK-3B with TDZD-8 attenuated hyperexcitability and restored HFCS-induced behavioural alterations.