Broad Kinase Inhibition Mitigates Early Neuronal Dysfunction in Tauopathy.

Tauopathies are a group of more than twenty known disorders that involve progressive neurodegeneration, cognitive decline and pathological tau accumulation. Current therapeutic strategies provide only limited, late-stage symptomatic treatment. This is partly due to lack of understanding of the molecular mechanisms linking tau and cellular dysfunction, especially during the early stages of disease progression. In this study, we treated early stage tau transgenic mice with a multi-target kinase inhibitor to identify novel substrates that contribute to cognitive impairment and exhibit therapeutic potential. Drug treatment significantly ameliorated brain atrophy and cognitive function as determined by behavioral testing and a sensitive imaging technique called manganese-enhanced magnetic resonance imaging (MEMRI) with quantitative R1 mapping. Surprisingly, these benefits occurred despite unchanged hyperphosphorylated tau levels. To elucidate the mechanism behind these improved cognitive outcomes, we performed quantitative proteomics to determine the altered protein network during this early stage in tauopathy and compare this model with the human Alzheimer's disease (AD) proteome. We identified a cluster of preserved pathways shared with human tauopathy with striking potential for broad multi-target kinase intervention. We further report high confidence candidate proteins as novel therapeutically relevant targets for the treatment of tauopathy. Proteomics data are available via ProteomeXchange with identifier PXD023562.

The effects of mild closed head injuries on tauopathy and cognitive deficits in rodents: Primary results in wild type and rTg4510 mice, and a systematic review.

In humans, the majority of sustained traumatic brain injuries (TBIs) are classified as 'mild' and most often a result of a closed head injury (CHI). The effects of a non-penetrating CHI are not benign and may lead to chronic pathology and behavioral dysfunction, which could be worsened by repeated head injury. Clinical-neuropathological correlation studies provide evidence that conversion of tau into abnormally phosphorylated proteotoxic intermediates (p-tau) could be part of the pathophysiology triggered by a single TBI and enhanced by repeated TBIs. However, the link between p-tau and CHI in rodents remains controversial. To address this question experimentally, we induced a single CHI or two CHIs to WT or rTg4510 mice. We found that 2× CHI increased tau phosphorylation in WT mice and rTg4510 mice. Behavioral characterization in WT mice found chronic deficits in the radial arm water maze in 2× CHI mice that had partially resolved in the 1× CHI mice. Moreover, using Manganese-Enhanced Magnetic Resonance Imaging with R1 mapping - a novel functional neuroimaging technique - we found greater deficits in the rTg4510 mice following 2× CHI compared to 1× CHI. To integrate our findings with prior work in the field, we conducted a systematic review of rodent mild repetitive CHI studies. Following Prisma guidelines, we identified 25 original peer-reviewed papers. Results from our experiments, as well as our systematic review, provide compelling evidence that tau phosphorylation is modified by experimental mild TBI studies; however, changes in p-tau levels are not universally reported. Together, our results provide evidence that repetitive TBIs can result in worse and more persistent neurological deficits compared to a single TBI, but the direct link between the worsened outcome and elevated p-tau could not be established.

Proteomic Techniques to Examine Neuronal Translational Dynamics.

Impairments in translation have been increasingly implicated in the pathogenesis and progression of multiple neurodegenerative diseases. Assessing the spatiotemporal dynamics of translation in the context of disease is a major challenge. Recent developments in proteomic analyses have enabled the resolution of nascent peptides in a short timescale on the order of minutes. In addition, a quantitative analysis of translation has progressed in vivo, showing remarkable potential for coupling these techniques with cognitive and behavioral outcomes. Here, we review these modern approaches to measure changes in translation and ribosomal function with a specific focus on current applications in the mammalian brain and in the study of neurodegenerative diseases.

Tau drives translational selectivity by interacting with ribosomal proteins.

There is a fundamental gap in understanding the consequences of tau-ribosome interactions. Tau oligomers and filaments hinder protein synthesis in vitro, and they associate strongly with ribosomes in vivo. Here, we investigated the consequences of tau interactions with ribosomes in transgenic mice, in cells, and in human brain tissues to identify tau as a direct modulator of ribosomal selectivity. First, we performed microarrays and nascent proteomics to measure changes in protein synthesis. Using regulatable rTg4510 tau transgenic mice, we determined that tau expression differentially shifts both the transcriptome and the nascent proteome, and that the synthesis of ribosomal proteins is reversibly dependent on tau levels. We further extended these results to human brains and found that tau pathologically interacts with ribosomal protein S6 (rpS6 or S6), a crucial regulator of translation. Consequently, protein synthesis under translational control of rpS6 was reduced under tauopathic conditions in Alzheimer's disease brains. Our data establish tau as a driver of RNA translation selectivity. Moreover, since regulation of protein synthesis is critical for learning and memory, aberrant tau-ribosome interactions in disease could explain the linkage between tauopathies and cognitive impairment.

Lrrk promotes tau neurotoxicity through dysregulation of actin and mitochondrial dynamics.

Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common cause of familial Parkinson disease. Genetics and neuropathology link Parkinson disease with the microtubule-binding protein tau, but the mechanism of action of LRRK2 mutations and the molecular connection between tau and Parkinson disease are unclear. Here, we investigate the interaction of LRRK and tau in Drosophila and mouse models of tauopathy. We find that either increasing or decreasing the level of fly Lrrk enhances tau neurotoxicity, which is further exacerbated by expressing Lrrk with dominantly acting Parkinson disease-associated mutations. At the cellular level, altering Lrrk expression promotes tau neurotoxicity via excess stabilization of filamentous actin (F-actin) and subsequent mislocalization of the critical mitochondrial fission protein dynamin-1-like protein (Drp1). Biochemically, monomeric LRRK2 exhibits actin-severing activity, which is reduced as increasing concentrations of wild-type LRRK2, or expression of mutant forms of LRRK2 promote oligomerization of the protein. Overall, our findings provide a potential mechanistic basis for a dominant negative mechanism in LRRK2-mediated Parkinson disease, suggest a common molecular pathway with other familial forms of Parkinson disease linked to abnormalities of mitochondrial dynamics and quality control, and raise the possibility of new therapeutic approaches to Parkinson disease and related disorders.

Designing antibodies against LRRK2-targeted tau epitopes.

Phosphorylation of the microtubule associated protein tau is an important modulator of its normal physiological functioning; however, it may also contribute to tau mis-folding and aggregation in neurodegenerative diseases, which are collectively termed tauopathies. As such, the investigations of tau phosphorylation and kinases that modify tau are important in trying to elucidate tau function and the mechanisms involved in the development of tauopathies. We have recently demonstrated that the putative tau kinase leucine-rich repeat kinase 2 is capable of phosphorylating tau at threonines 169 and 175 in vitro, and it has been previously shown that hyperphosphorylation at threonine 175 occurs in filamentous tau species from Alzheimer's brain tissue. These prior findings suggest that further studies of phosphorylation of tau at these epitopes may shed light on the pathogenesis of tauopathies. There is, however, a lack of tools available to analyze phosphorylation of tau at these sites. This study aimed to bridge that resource gap by generating monoclonal antibodies against tau phosphorylated at either threonine 169 or 175. While we did not succeed in generating a phospho-specific antibody, we did generate an antibody, MHT2, which is specific for human tau encompassing the threonine 169/175 epitope region. Immunostaining of transgenic rTg4510 mouse tissue as well as human tauopathy cases with MHT2 indicates that this antibody selectively detects cytoplasmic tau in the form of neurofibrillary tangles, and that it may have a further specificity pertaining to severity of disease progression, either because of phosphorylation or conformational bias.

Case Report of Extracorporeal Membrane Oxygenation and Aeromedical Evacuation at a Deployed Military Hospital.

The U.S. Military no longer maintains overseas extracorporeal membrane oxygenation (ECMO) capability for patients with severe lung injury including acute respiratory distress syndrome (ARDS). The authors present a case of severe ARDS at a military hospital in Afghanistan with limited capability for rescue therapies to include presentation, treatment, transport, and use of ECMO in the deployed military environment at one Role 3 medical facility. Lack of ECMO in the overseas environment is a significant gap in U.S. Military medical capability. The authors propose a novel solution, "ECMO packs," for prepositioning at strategic Role 3 facilities for early intervention in patients with severe lung injury to close this lethal and unnecessary capability gap.

Effects of two pre-workout supplements on concentric and eccentric force production during lower body resistance exercise in males and females: a counterbalanced, double-blind, placebo-controlled trial.

BACKGROUND: Pre-workout supplements purportedly enhance feelings of energy, reduce fatigue and improve exercise performance. The purpose of this study was to examine the performance effects of caffeinated and non-caffeinated multi-ingredient pre-workout supplements. METHODS: In a counterbalanced, double-blind, placebo-controlled design, eccentric and concentric force production during lower body resistance exercise on a mechanized squat device were assessed after supplement ingestion. Repetitions-in-reserve/RPE and subjective feelings of energy, focus and fatigue were also examined. Twenty-one resistance-trained adults (12 F, 9 M) completed three conditions in random order: caffeinated supplement, non-caffeinated supplement and placebo. Subjects were not informed of the presence of a placebo condition. Thirty minutes after supplement ingestion, a 3-repetition maximum test and 5 sets of 6 repetitions were completed using the squat device. Each repetition involved 4-s eccentric and concentric phases, and the force signal throughout each repetition was sampled from a load cell contained within the squat device. The scaled and filtered force signals were analyzed using customized software. Repeated measures analysis of variance and appropriate follow-up analyses were utilized to compare dependent variables, and relevant effect sizes (d) were calculated. RESULTS: Supplement or placebo ingestion led to similar subjective responses (p > 0.05). Energy (+8 to 44%; d = 0.3 to 0.8) and focus (+8 to 25%; d = 0.3 to 0.5) were acutely increased by supplement or placebo ingestion and decreased as the exercise session progressed. Fatigue was acutely decreased by supplement or placebo ingestion (-7 to 38%; d = -0.1 to -0.6) and increased as the exercise session progressed. Eccentric and concentric forces were unimproved by supplementation during the exercise sets for both sexes. In the non-caffeinated supplement condition only, maximal eccentric force production was lower during sets 3 to 5, as compared to set 1 (p < 0.05). Effect size data indicated that both the caffeinated and non-caffeinated supplements may contribute to small increases in concentric force production in males (+5 to 20%, d = 0.2 to 0.4 relative to placebo), but not females. CONCLUSIONS: As compared to placebo, caffeinated and non-caffeinated multi-ingredient pre-workout supplements failed to improve concentric and eccentric force production. In males, effect size data indicate a possible small benefit of supplementation on concentric force production, although this was not statistically significant. When resistance-trained subjects were unaware of the presence of a placebo, resistance exercise performance was similar regardless of whether a placebo or multi-ingredient supplement was ingested.

Adaptations Associated With an After-School Strength and Conditioning Program in Middle-School-Aged Boys: A Quasi-Experimental Design.

High-intensity strength and conditioning programs aimed at improving youth performance are becoming increasingly prevalent. The purpose of this study was to investigate the effects of a 16-week after-school strength and conditioning program on performance and body composition in middle-school-aged boys. Subjects in the training group (n = 16, mean age = 11.8 years) performed 90 minutes of supervised plyometric and resistance training twice weekly for 16 weeks. A group of control subjects (n = 9, age = 12.1 years) maintained their current activity levels. Sprint speed, 5-10-5 proagility, jump height, isometric peak torque of the leg extensors and flexors, and dual energy x-ray absorptiometry-derived body composition were examined during pretesting and posttesting. Data were analyzed by performing independent samples t-tests on the absolute change scores between groups. The primary findings were that the training intervention elicited significant improvements in 20-m sprint times (p = 0.03; mean change for training group = -0.17 seconds) and body-fat percentage (p = 0.03; 2.5% absolute improvement), the latter of which was a function of reduced fat mass (p = 0.06; -0.84 kg). Between-group differences were not noted for agility, jump height, lean mass, or strength measures; however, effect sizes generally showed greater improvements for the training group. In contrast to findings in longitudinal studies performed in collegiate athletes, sprint speed may be particularly adaptable during adolescence. In addition to potentially improving sport performance, high-intensity plyometric and resistance training programs offer the added benefit of improved body composition. These programs appear less effective for agility and jump performance and do not elicit substantial improvements in muscle mass above maturation.

Physiologically relevant factors influence tau phosphorylation by leucine-rich repeat kinase 2.

Hyperphosphorylation and aggregation of tau are observed in multiple neurodegenerative diseases termed tauopathies. Tau has also been implicated in the pathogenesis of Parkinson's disease (PD) and parkinsonisms. Some PD patients with mutations in the leucine-rich repeat kinase 2 (LRRK2) gene exhibit tau pathology. Mutations in LRRK2 are a major risk factor for PD, but LRRK2 protein function remains unclear. The most common mutation, G2019S, is located in the kinase domain of LRRK2 and enhances kinase activity in vitro. This suggests that the kinase activity of LRRK2 may underlie its cellular toxicity. Recently, in vitro studies have suggested a direct interaction between tubulin-bound tau and LRRK2 that results in tau phosphorylation at one identified site. Here we present data suggesting that microtubules (MTs) enhance LRRK2-mediated tau phosphorylation at three different epitopes. We also explore the effect of divalent cations as catalytic cofactors for G2019S LRRK2-mediated tau phosphorylation and show that manganese does not support kinase activity but inhibits the efficient ability of magnesium to catalyze LRRK2-mediated phosphorylation of tau. These results suggest that cofactors such as MTs and cations in the cellular milieu have an important impact on LRRK2-tau interactions and resultant tau phosphorylation.