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

Epigenetic modifications by polyphenolic compounds alter gene expression in the hippocampus

In this study, we developed an experimental protocol leveraging enhanced reduced representation bisulphite sequencing to investigate methylation and gene expression patterns in the hippocampus in response to polyphenolic compounds. We report that the administration of a standardized bioavailable polyphenolic preparation (BDPP) differentially influences methylated cytosine patterns in introns, UTR and exons in hippocampal genes. We subsequently established that dietary BDPP-mediated changes in methylation influenced the transcriptional pattern of select genes that are involved in synaptic plasticity. In addition, we showed dietary BDPP mediated changes in the transcriptional pattern of genes associated with epigenetic modifications, including members of the DNA methyl transferase family (DNMTs) and the Ten-eleven translocation methylcytosine dioxygenases family (TETs). We then identified the specific brain bioavailable polyphenols effective in regulating the transcription of DNMTs, TETs and a subset of differentially methylated synaptic plasticity-associated genes. The study implicates the regulation of gene expression in the hippocampus by epigenetic mechanisms as a novel therapeutic target for dietary polyphenols

Dietary polyphenols promote resilience against sleep deprivation-induced cognitive impairment by activating protein translation

Previous evidence has suggested that dietary supplementation with a bioactive dietary polyphenol preparation(BDPP)rescues impairment ofhippocampus-dependentmemory in amousemodelof sleepdeprivation (SD). In the current study, we extend our previous evidence and demonstrate that a mechanism by which dietary BDPP protects against SD-mediated cognitive impairment is via mechanisms that involve phosphorylation of the mammalian target of rapamycin complex 1 and its direct downstream targets, including the eukaryotic translation initiation factor 4E (eIF4E)-binding protein 1 (4E-BP1) and the ribosomal protein S6 kinase β-1 (p70S6K). In additional mechanistic studies in vitro, we identified the brain bioavailable phenolic metabolites derived from the metabolismof dietary BDPP that are responsible for the attenuation of SD-mediated memory impairments. On the basis of high-throughput bioavailability studies of brain bioavailablemetabolites after dietaryBDPP treatment,we found that select polyphenol metabolites [e.g., cyanidin-39-O-glucoside and 3-(39-hydroxyphenyl) propionic acid] were able to rescuemTORand p70S6K phosphorylation in primary cortico-hippocampal neuronal cultures, aswell as rescue 4E-BP1 phosphorylation in response to treatment with 4EGI-1, a specific inhibitor of eIF4E-eIF4G interaction. Our findings reveal a previously unknown role for dietary polyphenols in the rescue of SD-mediated memory impairments via mechanisms involving the promotion of protein translation

Forced Alternation (Y-Maze).

<p><b>A)</b> Scopolamine-HBr (10 mg/kg) or saline was administered by i.p. injection 30 min before the sample trial. A retrieval trial was conducted 30 min after the sample trial. The time spent in the novel arm during the first minute of the retrieval trial was analyzed. For both treatment groups (saline control: Ctrl; scopolamine: SCP), time spent in the novel arm was about 33%, with no statistically significant difference between groups (n = 9/group). <b>B)</b> In the second arm entry of the retrieval trial, saline-injected control mice entered the novel arm less often than scopolamine-treated mice (SCP). <b>C)</b> 129S6/Tg2576 mice (11–13 months) spent significantly less time in the novel arm during the retrieval trial and <b>D)</b> entered significantly less often into the novel arm (n = 19-20/group). The control has no error bar because all mice (100%) first went into the novel arm. Mice with less than 3 arm entries in the first minute were excluded from the analysis. Data are mean ± SEM; **<i>p <</i> 0.01; ***<i>p <</i> 0.001 (Student’s <i>t</i>-test for Fig 2A and 2C; Chi-squared test for Fig 2B and 2D).</p

A Comprehensive Behavioral Test Battery to Assess Learning and Memory in 129S6/Tg2576 Mice

<div><p>Transgenic Tg2576 mice overexpressing human amyloid precursor protein (hAPP) are a widely used Alzheimer’s disease (AD) mouse model to evaluate treatment effects on amyloid beta (Aβ) pathology and cognition. Tg2576 mice on a B6;SJL background strain carry a recessive <i>rd1</i> mutation that leads to early retinal degeneration and visual impairment in homozygous carriers. This can impair performance in behavioral tests that rely on visual cues, and thus, affect study results. Therefore, B6;SJL/Tg2576 mice were systematically backcrossed with 129S6/SvEvTac mice resulting in 129S6/Tg2576 mice that lack the <i>rd1</i> mutation. 129S6/Tg2576 mice do not develop retinal degeneration but still show Aβ accumulation in the brain that is comparable to the original B6;SJL/Tg2576 mouse. However, comprehensive studies on cognitive decline in 129S6/Tg2576 mice are limited. In this study, we used two dementia mouse models on a 129S6 background—scopolamine-treated 129S6/SvEvTac mice (3–5 month-old) and transgenic 129S6/Tg2576 mice (11–13 month-old)–to establish a behavioral test battery for assessing learning and memory. The test battery consisted of five tests to evaluate different aspects of cognitive impairment: a Y-Maze forced alternation task, a novel object recognition test, the Morris water maze, the radial arm water maze, and a Y-maze spontaneous alternation task. We first established this behavioral test battery with the scopolamine-induced dementia model using 129S6/SvEvTac mice and then evaluated 129S6/Tg2576 mice using the same testing protocol. Both models showed distinctive patterns of cognitive impairment. Together, the non-invasive behavioral test battery presented here allows detecting cognitive impairment in scopolamine-treated 129S6/SvEvTac mice and in transgenic 129S6/Tg2576 mice. Due to the modular nature of this test battery, more behavioral tests, e.g. invasive assays to gain additional cognitive information, can easily be added.</p></div

Behavioral Test Battery.

<p>Mice were tested in a behavioral test battery consisting of (top panel, from left to right): forced alternation (Y-maze), novel object recognition, Morris water maze, radial arm water maze, and spontaneous alternation (Y-maze) over a period of 8 weeks. Top panel: Behavioral tests conducted in the study. Middle panel: Timeline of tests. Bottom panel: Trials conducted per behavioral test. Tests were conducted in the order of increasing invasiveness. The spontaneous alternation test was performed last to maximize the interval to the forced alternation test, which utilized the same maze.</p

Spontaneous Alternation (Y-Maze).

<p><b>A)</b> Scopolamine-injected mice (SCP) and control (Ctrl) mice showed the same level of spontaneous alternation (n = 9-10/group). <b>B)</b> Scopolamine-treated mice had significantly more arm entries than control mice. <b>C)</b> 129S6/Tg2576 mice and WT mice performed showed the same level of spontaneous alternation (n = 16-19/group). <b>D)</b> The number of total arm entries between WT and 129S6/Tg2576 mice was not statistically different. Mice with less than 8 arm entries were excluded from the alternation analysis. Data are mean ± SEM; **<i>p</i> < 0.01 (Student’s <i>t</i>-test).</p

Novel Object Recognition.

<p><b>A)</b> Both control (Ctrl) and scopolamine-treated (SCP) mice showed no preference for the novel object over the familiar object (n = 8-9/group), but control mice showed a trend to interact more with the novel object (<i>p</i> = 0.466). <b>B)</b> Tg2576 mice showed a trend to interact less with the novel object (<i>p</i> = 0.257; n = 18-19/group). Mice with less than 7 sec of object interaction in either trial were excluded from the analysis. Data are mean ± SEM; Student’s <i>t</i>-test was used to calculate <i>p</i> values.</p

Radial Arm Water Maze.

<p><b>A)</b> Scopolamine-treated mice (SCP) made more errors at learning (trials 1–4) and remembering (trial 5) the hidden platform location than control mice (Ctrl) did (n = 9-10/group). <b>B)</b> Long-term (24 h) memory of mice was evaluated by averaging the entries into the previous target arm during the first trial of days 2–12. SCP mice entered into the last target arm significantly less than control mice. <b>C)</b> 129S6/Tg2576 mice made significantly more errors during the learning phase (trials 1–4) of the radial arm water maze test compared to WT control mice. <b>D)</b> The number of entries into the last target arm were not different between 129S6/Tg2576 mice and WT control mice (n = 19-20/group). Data are mean ± SEM; 6A and 6C: *<i>p</i> < 0.05; **<i>p</i> < 0.01; ***<i>p</i> < 0.001 (repeated measures ANOVA); 6B and 6D: ***<i>p</i> < 0.001 (repeated measures ANOVA was used for Fig 6A and 6C; Student’s t-test was used to calculated p values for Fig 6B and 6D).</p