bims-brabim Biomed News
on Brain bioenergetics and metabolism
Issue of 2022–02–06
thirty-one papers selected by
João Victor Cabral-Costa, University of São Paulo



  1. Alzheimers Dement. 2021 Dec;17 Suppl 3 e056456
       BACKGROUND: We reported previously that the neurosteroid allopregnanolone (Allo) promotes neural stem cell regeneration and differentiation, reverses neurogenic, metabolic and cognitive deficits and reduces Alzheimer's disease (AD) pathology in a mouse model of AD. To further investigate the cell-type specific mechanisms of Allo in regulating brain energy metabolism, we assessed the effect of Allo on mitochondrial bioenergetic profile and biogenesis in rat hippocampal astrocytes.
    METHOD: E18 rat hippocampal astrocyte were cultured for 10 days in DMEM:F12(1:1) with 10% FBS and then starved in 10% Charcoal stripped-FBS / DMEM:F12 for 24 hours before treatment with 100nM Allo or 0.001% Vehicle overnight. Upon completion of treatment, cells were subject to morphological, biochemical, metabolic and transcriptomic characterization of their mitochondrial phenotypes.
    RESULT: In primary hippocampal astrocytes, Allo significantly attenuates serum deprivation-induced bioenergetic deficits and oxidative stress by enhancing mitochondrial biogenesis and rebalancing mitochondrial dynamics. Allo treatment significantly enhances astrocytic mitochondrial biogenesis via Nrf1/Tfam signaling and reverses mitochondrial hyperfusion by elevating the ratio of mitochondrial fission protein Drp1 to the fusion protein Opa1. Functionally, Allo-induced improvement in bioenergetic function is coupled with reduced inflammasome activation in astrocytes.
    CONCLUSION: Outcomes of our findings further support the promising therapeutic effects of Allo against bioenergetic deficits that emerge in early phases of AD, with mitochondria being a major effector.
    DOI:  https://doi.org/10.1002/alz.056456
  2. Alzheimers Dement. 2021 Dec;17 Suppl 3 e057480
       BACKGROUND: Mitochondria regulate energy and metabolic homeostasis, with increasing evidence implicating mitochondrial dysfunction in Alzheimer's disease (AD) pathogenesis. Each mitochondrion contains multiple copies of the mitochondrial genome (mtDNA), with mtDNA copy number (mtDNAcn) been used as a surrogate measure of mitochondrial function. Here we evaluate the association of mtDNAcn with neuropathological diagnosis of AD and evaluate shared genetic etiology between AD and mtDNAcn.
    METHODS: We evaluated the association of mtDNAcn with a neuropathological diagnosis of AD in 1194 non-Hispanic white subjects (cases = 706, controls = 468) from three cohorts (ROSMAP, MSBB, Mayo) from the Accelerating Medicines Partnership Alzheimer's disease (AMP-AD). Relative mtDNAcn was estimated as the ratio of mtDNA to nuclear DNA using whole genome sequencing data from DNA isolated from brain tissue. Neuropathological AD was determined based on neuropathological burden of amyloid plaques and tangles. Logistic regression adjusting for mitochondrial haplogroup, age of death, sex, APOE, post-mortem interval and source tissue was used to evaluate the association of mtDNAcn with AD in each cohort separately and jointly in an inverse weighted fixed-effects meta-analysis (IVW FE). Additionally, we estimated the genetic correlation between mtDNAcn and AD, evaluated the association of a mtDNAcn polygenic risk score (PRS) with clinical AD (cases = 13312, controls = 13119), and conducted bidirectional two-sample Mendelian randomization to estimate the causal relationship between mtDNAcn and AD.
    RESULTS: Higher mtDNAcn was associated with a reduced risk of neuropathological AD (IVW FE: OR[95%CI] = 0.70 [0.58, 0.84]. mtDNAcn was not genetically correlated with AD (rg = 0.13 (0.16), p = 0.4) and a mtDNAcn PRS was not associated with clinical AD (OR[95%CI] = 1 [0.97, 1.03], p = 0.884). Mendelian randomization did not support a causal relationship between mtDNAcn and AD (OR[95%CI] = 0.93 [0.74, 1.14], p = 0.46).
    CONCLUSION: Elevated mtDNAcn estimated from brain tissue was associated with a reduced risk of neuropathological AD, suggesting that mitochondrial dysfunction is associated with AD pathogenesis. However, genetically predicted mtDNAcn estimated from peripheral blood was not associated with AD using genetically informed approaches. As such, further research is needed to determine if mitochondrial dysfunction causes, mediates, or is a by-product of AD pathogenesis.
    DOI:  https://doi.org/10.1002/alz.057480
  3. Cell Mol Life Sci. 2022 Feb 04. 79(2): 120
      Loss of neuronal polarity and missorting of the axonal microtubule-associated-protein TAU are hallmarks of Alzheimer's disease (AD) and related tauopathies. Impairment of mitochondrial function is causative for various mitochondriopathies, but the role of mitochondria in tauopathies and in axonal TAU-sorting is unclear. The axon-initial-segment (AIS) is vital for maintaining neuronal polarity, action potential generation, and-here important-TAU-sorting. Here, we investigate the role of mitochondria in the AIS for maintenance of TAU cellular polarity. Using not only global and local mitochondria impairment via inhibitors of the respiratory chain and a locally activatable protonophore/uncoupler, but also live-cell-imaging and photoconversion methods, we specifically tracked and selectively impaired mitochondria in the AIS in primary mouse and human iPSC-derived forebrain/cortical neurons, and assessed somatic presence of TAU. Global application of mitochondrial toxins efficiently induced tauopathy-like TAU-missorting, indicating involvement of mitochondria in TAU-polarity. Mitochondria show a biased distribution within the AIS, with a proximal cluster and relative absence in the central AIS. The mitochondria of this cluster are largely immobile and only sparsely participate in axonal mitochondria-trafficking. Locally constricted impairment of the AIS-mitochondria-cluster leads to detectable increases of somatic TAU, reminiscent of AD-like TAU-missorting. Mechanistically, mitochondrial impairment sufficient to induce TAU-missorting results in decreases of calcium oscillation but increases in baseline calcium, yet chelating intracellular calcium did not prevent mitochondrial impairment-induced TAU-missorting. Stabilizing microtubules via taxol prevented TAU-missorting, hinting towards a role for impaired microtubule dynamics in mitochondrial-dysfunction-induced TAU-missorting. We provide evidence that the mitochondrial distribution within the proximal axon is biased towards the proximal AIS and that proper function of this newly described mitochondrial cluster may be essential for the maintenance of TAU polarity. Mitochondrial impairment may be an upstream event in and therapeutic target for AD/tauopathy.
    Keywords:  Alzheimer's disease; Axon initial segment/AIS; Live-cell-imaging; Microtubule; Mitochondria; Mitochondriopathy; Neuron; Neuronal cell polarity; TAU; Tauopathy
    DOI:  https://doi.org/10.1007/s00018-022-04150-3
  4. Alzheimers Dement. 2021 Dec;17 Suppl 3 e054355
       BACKGROUND: Mitochondria are at the center of neural biogenergetics, and ApoE4 is the single most impactful risk factor for AD. We investigated the impact of ApoE on insulin sensitivity, on mitochondrial substrate utilization and bioenergetics. Persons with ApoE4 have reduced brain carbohydrate metabolism. To test for ApoE4 conferred neural mitochondrial metabolic differences, we constructed a novel stable-ApoE 2,3 and 4 N2a cell model, and tested ApoE's effects on Insulin sensitivity, and mitochondrial glucose, lipid and ketone oxidation.
    METHOD: Binding of ApoE isoforms E2, E3 and E4 to Insulin Receptor (IR) was measured by BLI and Co-IP, the impact of ApoE isoforms on mitochondrial glucose and lipid oxidation was measured by Seahorse.
    RESULT: ApoE3 was found to sensitize to insulin about 2-fold more potently than ApoE4. ApoE isoforms directly bind Insulin Receptor; the binding constants was in the range 200-300nM. Consistent with the previous insulin-sensitivity finding, ApoE3 caused a significant increase of the glycolytic rate and glucose oxidation relative to ApoE4. As there was no difference in oxidation of TCA cycle intermediates substrates in permeabilized cells, we infer ApoE3's glucose advantage is the result of increased insulin sensitivity. ApoE4 contributed a significant palmitate oxidation defect relative to ApoE2 and ApoE3. As this palmitate oxidation defect was observed in both mitochondria and cells it is likely to occur at or within mitochondria. We observed that the relative defect in ApoE4-dependent glucose and palmitate oxidation can be overcome by 5mM BHB. Thus, at the neural cell level, the metabolic defects contributed by ApoE 4 appear to be rescued by a ketogenic molecule, BHB, that requires neither insulin nor apolipoprotein particle to reach neural mitochondria and provide alternative metabolic support.
    CONCLUSION: ApoE4 confers 'double trouble' in mitochondrial glucose and lipid oxidation. ApoE4 confers a defect in mitochondrial lipid oxidation relative to all other isoforms. Simultaneously, ApoE4 lacks the benefit in glucose oxidation conferred by ApoE3, which appears to be driven by the reduced insulin sensitization potency of ApoE4. We also find that BHB can be an alternative source of neural bioenergy that enters mitochondria directly and thus is not affected by ApoE4 'double trouble'.
    DOI:  https://doi.org/10.1002/alz.054355
  5. Alzheimers Dement. 2021 Dec;17 Suppl 3 e051539
       BACKGROUND: Familial hypercholesterolemia (FH) is a genetic disorder caused by low-density lipoprotein receptor (LDLr) dysfunction, resulting in elevated plasma cholesterol levels. Previous reports have shown an interplay between LDLr and amyloid-β (Aβ) metabolism, a peptide linked to Alzheimer's disease. Indeed, LDLr-/- mice are more vulnerable to the deleterious memory impact induced by Aβ. Here, we investigated whether the gene expression of proteins involved in Aβ metabolism and Aβ content is altered in adult or middle-aged LDLr-/- mice brains. Also, we investigated neuroinflammation as well as neuronal and synaptic damage.
    METHOD: Young adult (3-month-old) and middle-aged (14-month-old) male C57BL/6 wild-type (WT) and LDLr-/- mice were first submitted to the Morris water maze test. After spatial memory assessment, the Aβ1-42 levels and gene expression of proteins involved in Aβ synthesis were evaluated in the prefrontal cortex (PFC) and hippocampus of 3 and 14-months-old WT and LDLr-/- mice. We also assessed the apoptosis signaling, levels of synaptic proteins, and Iba-1 immunoreactivity (a marker for microglia) in the experimental groups' brain structures.
    RESULT: LDLr-/- mice presented spatial memory impairment, which was more severe in middle-aged animals, which was not associated with altered expression of proteins involved in Aβ processing and Aβ1-42 levels in either hippocampus or PFC. We further found that the expression of the apoptotic protein Bax was increased in both the PFC and hippocampus of 3- and 14-month-old LDLr-/- mice. LDLr-/- mice presented increased immunoreactivity for activated caspase-3 in the neurons of the PFC and hippocampus. We also observed a reduction in immunocontent of PSD 95 in the hippocampus of 3-month-old LDLr-/- mice. Moreover, synaptophysin immunocontent was decreased in the middle-age LDLr-/- mice hippocampi. In addition, we observed that LDLr-/- mice displayed increased immunoreactivity for Iba-1 in the PFC already at 3 months of age and in the hippocampus at middle-age. Finally, we found that LDLr -/- at middle-age exhibited microglial morphological changes related to their activated state in the PFC.
    CONCLUSION: Cognitive impairments in LDLr-/- mice were associated with exacerbation of neuronal apoptosis, synaptic dysfunction, and microglial activation in brain regions related to memory formation, but not with significant changes in Aβ processing or levels.
    DOI:  https://doi.org/10.1002/alz.051539
  6. Alzheimers Dement. 2021 Dec;17 Suppl 3 e054315
       BACKGROUND: Astrocytes contribute to the normal function and maintenance of the health of the central nervous system (CNS). The abnormal behavior of astrocytes can lead to a major alteration in neuronal function contributing to the pathogenesis of several neurological diseases, including Alzheimer's disease (AD). Dysfunctional astrocytes have been linked to the onset and development of overreacting inflammation in CNS tissue which is a major contributing factor to the onset and progression of AD. We hypothesize that by modulating mitochondrial activity and cell metabolism using antioxidants, key characteristics of astrogliosis could be regulated. Mainly, we believe that some antioxidants can help to regulate astrocyte response and promote their return back to a quiescent state.
    METHODS: In this work, we explored the use of two antioxidant molecules (A1 and A2) as a way to regulate the metabolic activity, proliferation, and overreactive behavior of glial cells. Human astrocytes were cultured for five days in growth media supplemented with 0, 25, or 50 μM A1 or A2. Changes in DNA levels were used to calculate the doubling time of the cell population. DNA levels were quantified using PicoGreen dsDNA quantitation assay. Astrocytes were stained with JC-1 and fluorescence readings were taken to evaluate mitochondrial activity. Changes in metabolic activity were evaluated using the Vibrant Metabolic Assay and Adenosine Triphosphate (ATP) production.
    RESULTS: Quantitative assessments demonstrate that increasing levels of A1 significantly reduce human astrocyte proliferation while the addition of A2 at the chosen concentrations does not appear to have a significant effect on cell proliferation. The preliminary data for ATP production and metabolic activity illustrates that A1 appears to decrease cell metabolic activity which may lead to lower cell proliferation. In addition, quantitative analysis of the JC-1 mitochondrial staining suggests that the addition of A1 does not significantly impact mitochondrial membrane potential while increasing levels of A2 enhanced the polarization of the mitochondrial membrane.
    CONCLUSION: The modulation of mitochondrial and metabolic activity using antioxidant molecules may be used as a viable strategy to regulate the overreactive proliferative response of human astrocytes during astrogliosis.
    DOI:  https://doi.org/10.1002/alz.054315
  7. Alzheimers Dement. 2021 Dec;17 Suppl 3 e055430
       BACKGROUND: Previous studies suggested that individuals with a maternal history of Alzheimer's Disease (AD) are at higher risk of developing AD than individuals with a paternal history of AD. One could suggest that intra-uterine interactions might be responsible for elevating this risk. In this context, AD rodent models are highly suited for improving our understanding of this matter since animals reach adulthood in a few months. Here, we aimed at investigating changes in memory-related processes and brain metabolism on the offspring born to transgenic mothers harboring human APP/PS1 mutations. We hypothesized that offspring born to AD-transgenic mothers will present early memory and brain glucose metabolism changes.
    METHOD: Rats born to F344-AD (Tg-AD) mothers and wild-type fathers (WT), the maternal AD group, and rats born to WT mothers and Tg-AD fathers, the paternal AD group, were evaluated in two different time-points: ∼5.5, and ∼9.5 months. To assess spatial working memory, spontaneous alternation behavior was evaluated in the Y-maze test. Furthermore, micro-PET [18 F]FDG was used to assess brain glucose metabolism (SUVr normalization, pons as reference region).
    RESULT: Y-maze test demonstrated that rats born to transgenic mothers presented memory disturbances, indexed by spontaneous alternation, at ∼5.5 months (p = 0.0028, Figure 1A) and ∼9.5 months (p = 0.0062, Figure 1B), while rats born to WT mothers and Tg-AD fathers exhibited a decline in spontaneous alternation only at ∼9.5 months (p = 0.0181, Figure 1B). No changes were found in brain glucose metabolism at ∼5.5 months for both groups. However, rats born to AD-Tg mothers present brain glucose hypermetabolism at ∼9,5 months.
    CONCLUSION: Our findings demonstrate that rats born to AD-Tg mothers harboring APP/PS1 mutations present cognitive decline and brain glucose metabolism abnormalities earlier than those born to WT mothers. Further studies are needed to understand the biological basis behind this phenomenon.
    DOI:  https://doi.org/10.1002/alz.055430
  8. Alzheimers Dement. 2021 Dec;17 Suppl 3 e054793
       BACKGROUND: Impairment of brain glucose metabolism has been frequently described in Alzheimer's disease (AD). Moreover, the strongest predictor of the lifetime incidence of AD is the ε4 allele of APOE, a protein involved in lipid metabolism. These connections between AD and metabolism provide motivation to perform an in-depth metabolic profiling of human brain tissue for different stages of AD pathophysiology.
    METHOD: Brain tissue samples were obtained from the Religious Orders Study and Memory and Aging Project (ROS/MAP) at the Rush Alzheimer's Disease Center. Furthermore, ROS/MAP collects extensive phenotyping of the participants' cognitive trajectories as well as postmortem pathology. Metabolic profiling was performed on Metabolon's untargeted platform, yielding 1,055 quantified metabolites. Generalized linear models with appropriate linkage functions for continuous or categorical AD-related phenotypes were used to discover the association of metabolic profiles with AD-related phenotypes, such as amount of amyloid and tangles in brain, global burden of pathology, NIA-Reagan score, diagnosis (derived from Braak and CERAD scores), clinical diagnosis at the time of death, global cognition assessed during the visit before death, estimated decline of global cognition over lifetime. The models included confounder correction for age, gender, body mass index, years of education, post mortem interval, number of APOEε4 alleles, and medications.
    RESULT: We found 263 metabolites to be significantly associated (adjusted p-value <0.05) with one of the AD phenotypes. 137 of these metabolites were significantly associated with three or more phenotypes. Of these, nine could be replicated using an independent autopsy cohort from Mayo Clinic, five could also be replicated using a published study based on Baltimore Longitudinal Study of Aging cohort. The associated metabolites are involved in various metabolic processes known to be involved in AD pathogenesis, such as amino acid metabolism, urea cycle, and polyamine metabolism. In addition, we have identified several novel associations that could uncover the interdependence of different AD-associated metabolic processes.
    CONCLUSION: We have generated a comprehensive landscape of AD-associated metabolites and associated processes. These will be instrumental to fill the gap in our understanding of the metabolic components of AD pathophysiology.
    DOI:  https://doi.org/10.1002/alz.054793
  9. STAR Protoc. 2022 Mar 18. 3(1): 101120
      Mitochondrial electron transport chain (ETC) dysfunction elevates the NADH/NAD+ ratio to cause metabolic derangements. Here we describe a protocol to measure the NADH/NAD+ ratio and analyze the rewiring of glucose metabolism using [4-2H]-glucose, [3-2H]-glucose, and [U-13C]-glucose in ETC-inhibited human cancer cells. We also describe a protocol to analyze the NADH/NAD+ ratio-sensitive metabolites in mouse plasma and mouse liver following phenformin treatment. These protocols comprehensively analyze the metabolic derangements resulting from increased NADH/NAD+ ratio in in vitro and in vivo models. For complete details on the use and execution of this profile, please refer to Liu et al. (2021).
    Keywords:  Cell Biology; Cell culture; Cell-based Assays; Mass Spectrometry; Metabolism; Model Organisms
    DOI:  https://doi.org/10.1016/j.xpro.2021.101120
  10. Mol Cell Biochem. 2022 Jan 31.
      In the present study we have shown that treatment of SH-SY5Y cells with either thapsigargin or tunicamycin is associated with a significant decrease in ROUTINE and ATP-coupled mitochondrial respiration as well as a decrease in spare and maximal respiratory capacity. We have also shown that treating cells with either thapsigargin or tunicamycin is associated with significant changes in mitochondrial membrane potential (ΔΨm) generation, which is mainly associated with the reversal of the succinyl-CoA ligase reaction and a decreased activity of complex II. Despite the induction of endoplasmic reticulum (ER) specific unfolded protein response (UPR), as documented by increased expression of HRD1, ER stress did not induce mitochondrial UPR since the expression of both mitochondrial protease LONP1 and mitochondrial chaperone HSP60 was not significantly altered. Inhibition of IRE1α ribonuclease with STF-083010 did not protect the SH-SY5Y cells from ER stress-induced mitochondrial dysfunction. STF-083010 itself had significant impact on both mitochondrial respiration and generation of ΔΨm, which has mainly been associated with the uncoupling of respiratory chain from ATP synthesis.
    Keywords:  Endoplasmic reticulum stress; Mitochondrial dysfunction; Parkinson’s disease; Unfolded protein response
    DOI:  https://doi.org/10.1007/s11010-021-04344-6
  11. Alzheimers Dement. 2021 Dec;17 Suppl 3 e054287
       BACKGROUND: AMP-activated protein kinase (AMPK) is a cellular energy censor that has been implicated in Alzheimer's disease (AD). The catalytic α isoforms (α1 and α2) of AMPK are normally expressed in a homeostatic equilibrium but are differentially expressed in AD brains, with α2 expression significantly decreased compared to controls. AMPK dysregulation may be an early feature of AD that promotes accelerated cognitive decline through its roles in protein synthesis regulation, metabolism, autophagy, and mitochondrial biogenesis. The early loss of the α2 isoform may exacerbate or accelerate disease progression in the preclinical stage.
    METHOD: AMPKα2 was conditionally knocked down in excitatory forebrain neurons in the Tg19959 AD mouse model to create a heterozygous AMPKα2/Tg19959 double mutant. At 3-5 months of age, before cognitive impairments become evident, the mice were subjected to the Morris Water Maze (MWM) and Novel Object Recognition (NOR) task that measures hippocampus-dependent spatial learning and memory. In addition, Golgi staining, immunohistochemistry, and transmission electron microscopy of the hippocampus were performed.
    RESULT: The double mutant AMPKα2/Tg19959 mice showed significant memory impairments in both the MWM and NOR task compared to wild type, Tg19959, and α2/Cre littermates. The AMPKα2/Tg19959 double mutant mice also showed significant decreases in post-synaptic density size and in the number of polyribosomes. Differences in spine morphology and amyloid plaque deposition were also observed.
    CONCLUSION: AMPKα2 knockdown in a "pre-symptomatic" AD mouse model leads to accelerated cognitive deficits and disruption of protein synthesis regulatory mechanisms. Taken together, these findings suggest that reduction of AMPKα2 may exacerbate or accelerate cognitive decline and AD pathogenesis. This finding could provide a future therapeutic target for the treatment of AD.
    DOI:  https://doi.org/10.1002/alz.054287
  12. Glia. 2022 Feb 01.
      Maintenance of constant brain pH is critically important to support the activity of individual neurons, effective communication within the neuronal circuits, and, thus, efficient processing of information by the brain. This review article focuses on how glial cells detect and respond to changes in brain tissue pH and concentration of CO2 , and then trigger systemic and local adaptive mechanisms that ensure a stable milieu for the operation of brain circuits. We give a detailed account of the cellular and molecular mechanisms underlying sensitivity of glial cells to H+ and CO2 and discuss the role of glial chemosensitivity and signaling in operation of three key mechanisms that work in concert to keep the brain pH constant. We discuss evidence suggesting that astrocytes and marginal glial cells of the brainstem are critically important for central respiratory CO2 chemoreception-a fundamental physiological mechanism that regulates breathing in accord with changes in blood and brain pH and partial pressure of CO2 in order to maintain systemic pH homeostasis. We review evidence suggesting that astrocytes are also responsible for the maintenance of local brain tissue extracellular pH in conditions of variable acid loads associated with changes in the neuronal activity and metabolism, and discuss potential role of these glial cells in mediating the effects of CO2 on cerebral vasculature.
    Keywords:  acid-base balance; carbon dioxide; pH; respiratory circuits
    DOI:  https://doi.org/10.1002/glia.24152
  13. Front Mol Biosci. 2021 ;8 806650
      Noise exposure causes noise-induced hearing loss (NIHL). NIHL exhibits loss of inner ear sensory hair cells and is often irreparable. Although oxidative stress is involved in hearing loss, the complex mechanisms involved in NIHL are unclear. Hypoxia-inducible factor 1α (HIF-1α) has been suggested to be essential for protecting sensory hair cells. Additionally, it has been shown that ROS is involved in modulating the stability of HIF-1α. To investigate the NIHL pathogenesis, we established a tert-butyl hydroperoxide (t-BHP)-induced oxidative stress damage model in hair-like HEI-OC1 cells and an NIHL model in C57BL/6 mice. Protein and mRNA expression were determined, and biochemical parameters including reactive oxygen species (ROS) accumulation, glucose uptake, adenosine triphosphat (ATP) production, and mitochondrial content were evaluated. In HEI-OC1 cells, t-BHP induced ROS accumulation and reduced mitochondrial content and oxygen consumption, but the ATP level was unaffected. Additionally, there was increased glucose uptake and lactate release along with elevated expression of HIF-1α, glucose transporter 1, and several glycolytic enzymes. Consistently, noise trauma induced oxidative stress and the expression of HIF-1α and glycolytic enzymes in mice. Thus, we concluded that ROS induced HIF-1α expression, which promoted glycolysis, suggesting a metabolic shift maintained the ATP level to attenuate hair cell damage in NIHL.
    Keywords:  HIF-1α; energy metabolism; glycolysis; mitochondria; noise-induced hearing loss; oxidative stress; sensory hair cells
    DOI:  https://doi.org/10.3389/fmolb.2021.806650
  14. Alzheimers Dement. 2021 Dec;17 Suppl 3 e049489
       BACKGROUND: Fatty acid-binding protein 3 (FABP3) levels in the human cerebrospinal fluid elevate at early phases of Alzheimer's disease (AD), which is associated with reduced FABP3 levels in the brain and the onset of cognitive deterioration. However, it is unclear if reduced FABP3 levels in the brain can attribute to AD-like symptoms and pathology.
    METHOD: The cognitive function of FABP3 knockout (KO) and wildtype (WT) mice (15-week-old females) has been assessed using a battery of behavioural assessment tools, and the brain cytokines were subsequently assessed. Astrocytes were isolated from FABP3 KO and WT mice and cell metabolism and genes associated with metabolism and cytokines were examined.
    RESULT: FABP3 KO mice displayed deficits in short-term spatial memory (Fig 1A, 1B) and working memory (Fig 1C), which is associated with elevated IL-1β and TNF-α levels in the brain (Fig 1D). A significant decrease in glucose cellular uptake (Fig 2A) and metabolism (Fig 2B) and increase in fatty acid oxidation (Fig 2C) was demonstrated in FABP3 KO in relative to WT astrocytes. Genes responsible for fatty acid oxidation (carnitine palmitoyltransferase 1A) and transport (CD36) are upregulated in FABP3KO astrocytes (Fig 2D). In addition, IL-1β and TNFα mRNA in FABP3KO astrocytes increases by 3-fold and 4-fold, respectively (Fig 2D).
    CONCLUSION: This study demonstrates for the first time that FABP3 genetic knockout modifies astrocyte activities, which leads to AD-like pathology in mouse including elevated pro-inflammatory cytokine levels in the brain and cognitive dysfunction.
    DOI:  https://doi.org/10.1002/alz.049489
  15. Neurochem Int. 2022 Jan 29. pii: S0197-0186(22)00019-5. [Epub ahead of print] 105294
      It is known that brain energy metabolites such as ATP are quickly depleted during postmortem ischemia; however, a comprehensive assessment on the effects of preceding hypercapnia/ischemia and the dissection process on the larger brain metabolome remains lacking. This study sought to address this unknown by measuring aqueous metabolites impacted by hypercapnia/ischemia and brain dissection using Nuclear Magnetic Resonance. Metabolites were measured in rats subjected to 1) high energy head-focused microwave irradiation (control group); 2) CO2-induced hypercapnia/ischemia followed by immediate microwave irradiation; 3) CO2 followed by decapitation and then microwave irradiation ∼6.4 minutes later, to simulate a postmortem interval equivalent to typical dissection times; and 4) CO2-induced hypercapnia/ischemia followed by dissection within ∼6 minutes (no microwave fixation) to test the effects of brain dissection on the metabolome. Compared to microwave-irradiation, concentrations of high-energy phosphate metabolites and glucose were significantly reduced, while β-hydroxybutyrate and lactate were increased in rats subjected to all other treatments. Several amino acids and neurotransmitters (GABA) increased by hypercapnia/ischemia and dissection. Sugar donors involved in glycosylation decreased and nucleotides decreased or increased following hypercapnia/ischemia and dissection. sn-Glycero-3-phosphocholine decreased and choline increased in all groups relative to controls, indicating postmortem changes in lipid turnover. Antioxidants increased following hypercapnia/ischemia but decreased to control levels following dissection. This study demonstrates substantial post-mortem changes in brain energy and glycosylation pathways, as well as protein, nucleotide, neurotransmitter, lipid, and antioxidant turnover due to hypercapnia/ischemia and dissection. Changes in phosphate donors, glycosylation and amino acids reflect post-translational and protein degradation processes that persist post-mortem. Microwave irradiation is necessary for accurately capturing in vivo metabolite concentrations in brain.
    Keywords:  Dissection; Irradiation; Ischemia; Metabolomics; Microwave fixation
    DOI:  https://doi.org/10.1016/j.neuint.2022.105294
  16. Sci Rep. 2022 Feb 02. 12(1): 1820
      Numerous emotional and cognitive processes mediated by the hippocampus present differences between sexes and can be markedly influenced by hormonal status in males and females of several species. In rodents, the dorsal hippocampus (dHPC) is known to contribute to the rapid antidepressant actions of the NMDA receptor antagonist ketamine. We and others have demonstrated a greater sensitivity to the fast-acting antidepressant ketamine in female versus male rats that is estrogen- and progesterone-dependent. However, the underlying mechanisms remain unclear. Using an acute low dose (2.5 mg/kg) of ketamine that is behaviorally effective in female but not male rats, a label-free phosphoproteomics approach was employed to identify ketamine-induced changes in signaling pathway activation and phosphoprotein abundance within the dHPC of intact adult male rats and female rats in either diestrus or proestrus. At baseline, males and females showed striking dissimilarities in the dHPC proteome and phosphoproteome related to synaptic signaling and mitochondrial function-differences also strongly influenced by cycle stage in female rats. Notably, phosphoproteins enriched in PKA signaling emerged as being both significantly sex-dependent at baseline and also the primary target of ketamine-induced protein phosphorylation selectively in female rats, regardless of cycle stage. Reduced phosphoprotein abundance within this pathway was observed in males, suggesting bi-directional effects of low-dose ketamine between sexes. These findings present biological sex and hormonal milieu as critical modulators of ketamine's rapid actions within this brain region and provide greater insight into potential translational and post-translational processes underlying sex- and hormone-dependent modulation of ketamine's therapeutic effects.
    DOI:  https://doi.org/10.1038/s41598-022-05937-x
  17. Alzheimers Dement. 2021 Dec;17 Suppl 3 e052606
       BACKGROUND: Astrocytes respond dramatically to pathological alterations in the brain including Alzheimer's disease (AD) through a highly heterogeneous process called astrocyte reactivity. Astrocyte reactivity is characterized by changes in morphological, transcriptional and biochemical levels. The reactive astrocytes secrete a plethora of cytokines that regulate neuronal health through disease stages. Recently, we showed that oligomeric Aβ1-42 (Aβ) treatment to primary astrocytes induced an early reactivity and detected tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) as the major candidate in the astrocyte secretome that promoted neuronal viability against Aβ toxicity. Moreover, in Aβ-injected rats, TIMP-1 recovered cognitive functions (Brain Behav Immun. 2020 87:804-819). However, the mechanisms underlying TIMP-1's neuroprotective role remained to be revealed which we aim to achieve through our present study.
    METHOD: Primary cortical neurons were co-treated with Aβ and recombinant TIMP-1, and Akt phosphorylation at different sites and downstream protein levels were checked by western blotting and reconfirmed by immunofluorescence studies. Co-localization and immunoprecipitation studies were performed to determine TIMP-1's binding partner/s. Subsequently, TIMP-1 was stereotactically injected in 6-month old 5xFAD mice and cognitive behaviors were assessed through locomotion, passive avoidance, fear conditioning and elevated plus maze tests. Immunohistochemical analyses and western blotting were employed to analyze pathway inductions in vivo.
    RESULT: In neurons, TIMP-1 activated Akt by phosphorylating at two different residues: S473 and Thr308. Akt S473 phosphorylation ameliorated impaired autophagy flux and downregulated Aβ-induced pro-apoptotic protein levels by inhibiting FoxO3a in cultured neurons and in cortex and hippocampus of 5xFAD mouse. Subsequently, CD63 was detected as the binding partner of TIMP-1 on the neuronal surface. TIMP-1 treatment recovered the cognitive deficits in 5xFAD mice. Synaptic proteins, SNAP-25 and PSD-95, expressions were increased following TIMP-1 treatment in 5xFAD. This was consistent with Akt thr308 phosphorylation and downstream GSK3β phosphorylation-mediated inhibition, implicated in improving synaptic plasticity.
    CONCLUSION: Data show that TIMP-1 regulates differential phosphorylation of Akt that is potentially central to regulating autophagy, apoptotic pathways and synaptic plasticity in models of AD resulting in cognitive recovery. We highlight that TIMP-1 mediates its neuroprotective role by binding to CD63 and propose it as a primary candidate in astrocyte-secreted cytokine-mediated AD therapy.
    DOI:  https://doi.org/10.1002/alz.052606
  18. Alzheimers Dement. 2021 Dec;17 Suppl 3 e051164
       BACKGROUND: γ-secretase activity is enriched at mitochondria-associated ER membranes (MAMs),a lipid-raft subdomain of the ER, where APP fragment C99 is delivered for its cleavage. Moreover, γ-secretase activity deficiency linked to AD mutations causes C99 accumulation at MAM, resulting in the upregulation of MAM activities such as sphingomyelin turnover and cholesterol esterification. We now explore whether the interaction of C99 with cholesterol could explain the link between C99 accumulation at MAM and the lipid abnormalities found in AD.
    METHOD: Using cellular models of C99 accumulation versus a cholesterol-binding deficient C99, we assessed cholesterol uptake dynamics and sub-cellular distribution along with MAM-regulated functionalities. Also, we used a photo click activable cholesterol analog to study the proteins interacting with cholesterol at MAM in both conditions. Finally, we compare membrane permeability dynamics in neuronal models of Alzheimer's disease.
    RESULT: C99 accumulation at MAM caused an increase in cholesterol uptake, cholesterol esterification, phospholipid synthesis and sphingomyelinase activity accompanied by a sub cellular redistribution of cholesterol to endolysosomes and ER, while C99-defective in cholesterol binding showed no changes in these activities. Moreover, when the cholesterol-interacting proteome of both conditions was compared, C99 caused a huge change/recruitment at MAM, especially of enzymes involved in lipid metabolism. ACSL4 (Acyl-CoA Synthetase Long Chain Family Member 4) activation was detected as an important player in the lipidome changes associated with C99 accumulation that led to membrane permeability alterations CONCLUSION: We report that the lipid alterations caused by pathogenic C99 accumulation are a consequence of an exacerbated uptake of extracellular cholesterol and mobilization towards MAM. The increase content of cholesterol at MAM driven by C99 might be responsible of the persistent activation of MAM and a subsequent alteration of the lipid metabolism since a cholesterol-binding deficient C99 fails to increase MAM activities and cholesterol trafficking. We also point to ACSL4 activation as a possible molecular switch for the permeability changes found in cellular models of AD.
    DOI:  https://doi.org/10.1002/alz.051164
  19. Alzheimers Dement. 2021 Dec;17 Suppl 3 e057845
       BACKGROUND: We previously developed a temporal model for unobserved molecular changes occurring during late-onset Alzheimer's Disease (AD) for RNA-Seq data. Here we apply this method to proteomics data from an AD case-control cohort and show that the model's estimate of disease progression is associated with clinical and neuropathological endpoints, and identify changes in the proteome across stages of AD.
    METHODS: Manifold learning defines an order across samples based on their similarity of expression. This ordering estimates pseudotime, an inference of molecular disease progression, quantitatively measured as the distance of each sample from the start of the inferred trajectory. We applied this approach to TMT-proteomics data from human postmortem brain samples from the ROS/MAP study within the Accelerating Medicine Partnership-AD consortia (AMP-AD) (N=400). We examined associations between pseudotime and neuropathological and clinical endpoints. We performed differential expression analysis on tree branches to identify progression-specific pathways, and used gene set enrichment analysis to identify significant GO terms from a set that have been curated into 15 distinct AD-relevant biological domains.
    RESULTS: Pseudotime estimates were significantly associated with LOAD status (females, p=3.67x10-7; males, p=1.03x10-3), such that "early" (low pseudotime) samples are enriched for controls, and "late" (high pseudotime) samples are enriched for cases. Pseudotime was associated with cognitive diagnosis (females, p=2.46x10-5; males, p=1.14x10-3), Braak stage (females, p=1.48x10-5; males, p=1.52x10-3), and CERAD neuritic plaque score (females, p=1.17x10-11; males, p=1.32x10-4). Proteins involved in immune response were upregulated, while mitochondrial metabolism was downregulated, consistently across pseudotime. We observed differences between branches in biological domains encompassing vascular function, structural stabilization, lipid metabolism, apoptosis regulation, and epigenetics. Synaptic dysfunction was downregulated during mid-to-late pseudotime in women, but not in men.
    CONCLUSIONS: This approach to identifying biological changes associated with AD progression has now been applied to two data modalities, and despite substantial differences in data used to order trajectories, pseudotime was associated with multiple measures of AD progression in both studies. We also uncovered proteome-specific changes, providing opportunities to glean new insights about genetic drivers of AD across disease severity.
    DOI:  https://doi.org/10.1002/alz.057845
  20. Alzheimers Dement. 2021 Dec;17 Suppl 3 e054371
       BACKGROUND: The predominant genetic risk factor for late-onset Alzheimer's disease (AD) is the ε4 allele of apolipoprotein E (APOE4). APOE4-related risk of developing AD is modified by sex, with women showing greater risk than men. Regardless of APOE genotype, women exhibit a higher prevalence and lifetime risk of AD. Extensive human and animal model literatures indicate that women's risk for AD is also affected by the normal, age-related depletion of estrogens at menopause. For example, in mouse models, estrogen depletion by ovariectomy accelerates the onset and progression of AD-like neuropathology and cognitive impairment, whereas estrogen-based hormone therapies can partially prevent and or reverse these effects. Potential interactions between APOE4 genotype and estrogen status have not been well elucidated. To provide more insight into these issues, we conducted transcriptional analysis on hippocampi from adult female mice across APOE3 and APOE4 genotypes in the presence or absence of estradiol.
    METHOD: At 18-22 weeks of age, female mice homozygous for knock-in of human APOE3 or APOE4 were bilaterally ovariectomized (OVX) or sham-OVX and immediately implanted with a subcutaneous silastic capsule containing either vehicle or 0.5% 17β-estradiol (E2). Four weeks following surgery and hormone treatment, hippocampi were harvested and processed for polyA-enriched RNA-seq. Transcripts were mapped to a cDNA library of the mouse genome using kallisto (0.46.1). DEseq2 (1.28.1) normalized fold-changes were then used to estimate differential gene expression among groups.
    RESULT: RNA-seq analyses indicate significant differences in hippocampal transcriptional profiles across treatment groups. In particular, we found group differences in pathways related to synaptic integrity and plasticity, lipid metabolism and glucose metabolism. Interestingly, the strongest group differences were associated with APOE genotype whereas estrogen status yielded generally modest effects.
    CONCLUSION: Our results suggest that APOE genotype impacts the transcriptome in the female hippocampus to a larger degree than estrogen status. These findings provide novel insight into the independent and interactive neural effects of APOE4 and estrogen, which may provide opportunities for identification of therapeutic targets for AD.
    DOI:  https://doi.org/10.1002/alz.054371
  21. Alzheimers Dement. 2021 Dec;17 Suppl 3 e052443
       BACKGROUND: Alzheimer's disease (AD) is the most common age-related neurodegenerative disease, and an effective therapeutic strategy that promotes healthy aging may lower age-related risks of AD and ameliorate AD associated cognitive dysfunctions. There has been substantial interest in the application of ketogenic diets (KD) to manage neurological disorders associated with aging, and studies have also shown KDs started from early middle age improved cognition and longevity in mice. Thus, KDs might be used to reduce risk of cognitive declines at old age. KDs started later in life or intermittently administered may be more feasible and promote compliance in an older population. Therefore, this study sought to test if continuous or intermittent KDs started in late-middle-aged mice would improve measures of cognitive and motor function at advanced ages.
    METHOD: 18-month-old male C57BL/6J mice were randomly assigned to an isocaloric control (CD), ketogenic (KD), or intermittent ketogenic (IKD, 3 days of KD/week) diet. Continuous or intermittent ketosis was induced at a constant level of energy intake. At 20, 23, and 26 months of age, a panel of behavior tests were performed to assess cognitive (novel object recognition, Y-maze, and Barnes-maze) and motor (grid-wire hang, grip strength, open field, and rearing) functions.
    RESULT: Y-maze alternation rate was significantly higher for both IKD and KD mice at 23 months of age and for KD mice at 26 months indicating an improved working memory. 26-month-old KD mice also showed better spatial learning memory as measured by time spent in target quadrant in Barnes-maze. Improved motor endurance and strength was observed in aged IK and KD mice as tested by grid wire hang. A significantly increased composite score of all the behavior parameters was observed in KD mice at 26 months of age, and IK mice showed a trend toward increased score compared to CD mice.
    CONCLUSION: KD and IKD initiated in late-middle-aged mice improved cognition and motor endurance in aged mice. KD had a more potent effect on overall heath span in aged mice shown as a higher composite score of all the tests performed with IKD showing results intermediate to other diet groups.
    DOI:  https://doi.org/10.1002/alz.052443
  22. Alzheimers Dement. 2021 Dec;17 Suppl 3 e050377
       BACKGROUND: Chemobrain has recently been recognized as the most common neurological consequences following chemotherapy, particularly doxorubicin (Dox) treatment. In addition, the incidence of Alzheimer's disease (AD) increases in response to chemotherapy. Dox administration induced cognitive impairment via brain oxidative stress, neuroinflammation, mitochondrial dysfunction, cholinergic deficits, and cumulating neuronal apoptosis. Notably, donepezil (DPZ), an acetylcholinesterase inhibitor, has been proven to effectively attenuate various neuropathological conditions, particularly AD. However, the mechanistic insight regarding the protective effects of DPZ on cognition and brain pathologies in rats with doxorubicin-induced chemobrain has never been determined.
    METHOD: Fifteen male Wistar rats were randomized to receive either a 0.9% normal saline solution (Control; n=5) or 3 mg/kg Dox for 6 doses via intraperitoneal injection. Then, Dox-treated rats were divided into 2 groups to receive either vehicle (Dox, n=5) or DPZ (5 mg/kg: Dox+DPZ; n=5) orally, starting at 1st day of Dox for a 30-day duration. After completion of the treatment paradigm, cognitive function was evaluated using the novel object location (NOL) task. The brains were obtained for further molecular investigation.
    RESULT: Dox-treated rats exhibited impaired cognitive performance as presented by a decline in preference index in the NOL test. Loss of dendritic spines was observed following Dox treatment. Dox-treated rats showed brain mitochondrial dysfunction as indicated by increased mitochondrial reactive oxygen species (ROS) production (Figure 1) and imbalanced mitochondrial dynamics. Additionally, Dox induced neuroinflammation as demonstrated by increased TNF-α mRNA expression, together with enhanced microglial activation (Figure 1). Interestingly, Dox-treated rats developed potential amyloid lesions (Figure 1) and disrupted neurogenesis in the hippocampus which in turn led to neuronal cell death through both apoptosis and necroptosis. Strikingly, DPZ co-treatment completely restored cognitive function (Figure 1) by reducing dendritic spine loss, improving mitochondrial function, balancing mitochondrial dynamics, ameliorating neuroinflammation, decreasing Alzheimer's related proteins, preserving neurogenesis, and inhibiting apoptosis and necroptosis.
    CONCLUSION: These findings suggested that DPZ provided neuroprotection against Dox-induced chemobrain, which could be used for the therapeutic approach to prevent chemobrain in patients who received chemotherapy in the future.
    DOI:  https://doi.org/10.1002/alz.050377
  23. Alzheimers Dement. 2021 Dec;17 Suppl 3 e056647
       BACKGROUND: L-carnitine is present in the mammalian cells as free carnitine (FC) and acylcarnitine and the adult human brain contains almost 10% of long chain acylcarnitine. Acylcarnitines are functionally involved in β-oxidation of fatty acids and are also known for their role in neuroprotection. Levels of plasma acylcarnitines are known to decreased on aging. It is important to understand the association of acylcarnitines with cognitive impairment in Alzheimer's disease (AD).
    METHOD: We integrated the transcriptome data from 1000 post-mortem brain samples from ROS/MAP, Mayo clinic and Mount Sinai Brain bank cohort with the brain region-specific metabolic networks. We calculated the metabolic fluxes for the reactions in the model and identified those that showed differential fluxes in AD samples. We filtered the reactions that are involved in acylcarnitine synthesis and transport namely carnitine transport, fatty acid oxidation, citric acid cycle, and glutathione metabolism.
    RESULT: We found differences in metabolic fluxes for reactions involved in the acetylcarnitine transport to mitochondria (ACRNtm), carnitine palmitoyl transferase 1 and 2 (CPT1 and CPT2) as well as acyl-CoA dehydrogenase short and medium chain (ACADS, ACADM) located in mitochondria in AD samples. Using gene-based association analysis in participants of the AD Neuroimaging Initiative (ADNI) phases 1, GO and 2, we identified genetic variants linked to CPT1, CPT2, ACADM and ACADS genes suggested from the metabolic flux analysis.
    CONCLUSION: Our findings suggest that acylcarnitine synthesis and transport is altered in AD. Altered metabolism of short and medium chain acylcarnitines can be used as metabolic features of AD.
    DOI:  https://doi.org/10.1002/alz.056647
  24. Intensive Care Med Exp. 2022 Feb 04. 10(1): 4
       BACKGROUND: The use of norepinephrine in the case of life-threatening haemorrhagic shock is well established but widely discussed. The present study was designed to compare the effects of early norepinephrine treatment vs. no treatment on cerebral energy metabolism during haemorrhagic shock.
    METHODS: Twelve pigs were subjected to haemorrhagic shock, 4 in the control group and 8 in the norepinephrine (NE) group. Following a 60 min baseline period haemorrhagic shock was achieved by bleeding all animals to a pre-defined mean arterial blood pressure (MAP) of approximately 40 mm Hg. When mean arterial pressure had decreased to 40 mmHg NE infusion started in the treatment group. After 90 min, NE infusion stopped, and all pigs were resuscitated with autologous blood and observed for 2.5 h. During the experiment cerebral tissue oxygenation (PbtO2) was monitored continuously and variables reflecting cerebral energy metabolism (glucose, lactate, pyruvate, glutamate, glycerol) were measured by utilizing intracerebral microdialysis.
    RESULTS: All 12 pigs completed the protocol. NE infusion resulted in significantly higher MAP (p < 0.001). During the shock period lactate/pyruvate (LP) ratio group increased from 20 (15-29) to 66 (38-82) (median (IQR)) in the control group but remained within normal limits in the NE group. The significant increase in LP ratio in the control group remained after resuscitation. After induction of shock PbtO2 decreased markedly in the control group and was significantly lower than in the NE group during the resuscitation phase.
    CONCLUSION: NE infusion during haemorrhagic shock improved cerebral energy metabolism compared with no treatment.
    Keywords:  Cerebral metabolism; Haemorrhagic shock; Microdialysis; Norepinephrine; Trauma
    DOI:  https://doi.org/10.1186/s40635-022-00432-z
  25. Brain Res Bull. 2022 Jan 29. pii: S0361-9230(22)00017-X. [Epub ahead of print]181 97-108
      Subarachnoid hemorrhage (SAH) is a hemorrhagic stroke disease with high mortality and disability rates. Neurological recovery in early brain injury (EBI) after SAH is a crucial stage to reduce complications and improve the prognosis of patients. The mitochondrial unfolded protein response (UPRmt) is an essential mitochondrial damage repair process, that degrades aggresomes formed by misfolded proteins. UPRmt is a response to cellular stress and enhances mitochondrial homeostasis. GrpEL1 is a nucleotide exchange factor that assists mtHSP70 in nonnative folding proteins in mitochondria. However, the role of UPRmt and GrpEL1 after SAH is unclear. Western blot, Immunofluorescence, Aggresome staining, JC-1 staining were conducted to detect UPRmt after SAH in vivo and in vitro. The results showed that the UPRmt-related proteins HSP60 and mtHSP70 did not change in the EBI after SAH in vivo and in vitro but increased in the isolated mitochondria. In vitro primary neurons treated with oxyhemoglobin (OxyHb) achieved the same result as MG132 induction, increasing neuron protein aggresomes. The expression of GRPEL1 was unchanged in total protein and mitochondrial protein by Western blot. Co-immunoprecipitation (Co-IP) experiments showed that the GRPEL1-mtHSP70 complex decreased after OxyHb treatment. After GRPEL1 overexpression, the GRPEL1-mtHSP70 complex increased, while aggresome in neurons decreased. JC-1 showed an increased mitochondrial membrane potential, ATP content increased, and Western blot analysis revealed decreased cleaved-Caspase 9, suggesting improved mitochondrial function. In conclusion, the reduced GrpEL1-mtHSP70 complex is an essential factor affecting UPRmt in EBI after SAH. Increasing GrpEL1 promotes GrpEL1 and mtHSP70 binding, promoting the neuronal mitochondrial homeostasis, and might be an essential clinical intervention target for EBI after SAH.
    Keywords:  Aggresome; GrpEL1; HSP60; MtHSP70; Subarachnoid hemorrhage; UPRmt
    DOI:  https://doi.org/10.1016/j.brainresbull.2022.01.014
  26. Alzheimers Dement. 2021 Dec;17 Suppl 3 e050378
       BACKGROUND: Diabetes is a known risk factor for dementia, several factors are involved in the pathogenesis of diabetes including genetic predisposition, and high calorie diet consumption. We previously reported that long-term high-fat diet (HFD) consumption led to prediabetes, which associated with cognitive impairments. However, the effects of long-term HFD consumption on metabolic and brain function in genetically type 2 diabetes (T2DM) model have not been investigated. Therefore, we hypothesized that long-term HFD consumption aggravates metabolic and cognitive impairments in T2DM rats.
    METHOD: Sixteen Spontaneous Diabetic Torii (SDT) rats, which is a genetically engineered T2DM rats, and eight wild type (WT) rats were used. At the age of 8 weeks old, SDT rats were divided into two dietary groups either normal diet or HFD feeding for 12 weeks. WT rats were received a normal diet entire the study. At 20 weeks old, metabolic parameters, the hippocampal dependent cognitive function using novel object location (NOL) test, and hippocampal dendritic spine density were determined.
    RESULT: Our data demonstrated that, at 20 weeks old, SDT rats in both dietary groups exhibited their diabetic characters as indicated by hyperglycemia, hypoinsulinemia, and insulin insensitivity. HFD feeding did not aggravate diabetic status, but it significantly induced body weight gain, visceral fat deposition, hypertriglyceridemia, and hypercholesterolemia (as shown in Figure 1). Unexpectedly, hippocampal dependent cognitive impairments were not observed in both normal diet and HFD-fed SDT rats, when compared with WT rats (Figure 1). However, brain mitochondrial dysfunction and dendritic spine loss were observed in both dietary groups of SDT rats. Interestingly, the reduction of dendritic spine density was greater in HFD-fed SDT rats than normal diet-fed SDT rats (Figure 1).
    CONCLUSION: Long-term HFD consumption induced truncal obesity, dyslipidemia, and reduced dendritic spine density in SDT rats, but it did not affect hippocampal dependent cognitive function. It is possible that the duration of HFD feeding was insufficient to promote cognitive impairments in genetically T2DM rats.
    DOI:  https://doi.org/10.1002/alz.050378
  27. J Prev Alzheimers Dis. 2022 ;9(1): 54-66
       BACKGROUND: Ketone bodies have been proposed as an "energy rescue" for the Alzheimer's disease (AD) brain, which underutilizes glucose. Prior research has shown that oral ketone monoester (KME) safely induces robust ketosis in humans and has demonstrated cognitive-enhancing and pathology-reducing properties in animal models of AD. However, human evidence that KME may enhance brain ketone metabolism, improve cognitive performance and engage AD pathogenic cascades is scarce.
    OBJECTIVES: To investigate the effects of ketone monoester (KME) on brain metabolism, cognitive performance and AD pathogenic cascades in cognitively normal older adults with metabolic syndrome and therefore at higher risk for AD.
    DESIGN: Double-blinded randomized placebo-controlled clinical trial.
    SETTING: Clinical Unit of the National Institute on Aging, Baltimore, US.
    PARTICIPANTS: Fifty cognitively intact adults ≥ 55 years old, with metabolic syndrome.
    INTERVENTION: Drinks containing 25 g of KME or isocaloric placebo consumed three times daily for 28 days.
    OUTCOMES: Primary: concentration of beta-hydroxybutyrate (BHB) in precuneus measured with Magnetic Resonance Spectroscopy (MRS). Exploratory: plasma and urine BHB, multiple brain and muscle metabolites detected with MRS, cognition assessed with the PACC and NIH toolbox, biomarkers of AD and metabolic mediators in plasma extracellular vesicles, and stool microbiome.
    DISCUSSION: This is the first study to investigate the AD-biomarker and cognitive effects of KME in humans. Ketone monoester is safe, tolerable, induces robust ketosis, and animal studies indicate that it can modify AD pathology. By conducting a study of KME in a population at risk for AD, we hope to bridge the existing gap between pre-clinical evidence and the potential for brain-metabolic, pro-cognitive, and anti-Alzheimer's effects in humans.
    Keywords:  Alzheimer’s disease; Ketosis; cognition; extracellular vesicles; ketone ester; magnetic resonance spectroscopy; metabolic syndrome
    DOI:  https://doi.org/10.14283/jpad.2022.3
  28. Alzheimers Dement. 2021 Dec;17 Suppl 3 e054397
       BACKGROUND: Cardiovascular (CV) risk factors may potentiate cerebral amyloid angiopathy (CAA) pathology and neurovascular dysfunction, worsening neurodegeneration. The role of the mitochondria, key regulators of cell survival and death, in mediating cerebral endothelial cell (EC) dysfunction during CAA complicated with CV risk factors is unknown. Our lab has demonstrated that carbonic anhydrase inhibitors (CAi) reduce amyloid-induced cell death and mitochondrial dysfunction in EC. Here, we aim to understand the role of the mitochondria in the pathogenesis of mixed AD and vascular dementias. Furthermore, we seek to understand the effects of pan-CAi and CAi with high selectivity for mitochondria-localized isoforms (mCAi) in preventing the toxic effects of amyloid β (Aβ) and hyperhomocysteinemia (HHcy).
    METHODS: Human ECs were challenged with Aβ, Hcy, or the combination in the presence or absence of CAi. Cell death, mitochondrial function, and blood-brain barrier (BBB) resistance effects were evaluated.
    RESULTS: The presence of Aβ and/or HHcy induced EC apoptosis, mediated by death receptors activation and mitochondrial dysfunction, and attenuated by CAi and mCAi. BBB permeability was increased by both Aβ and HHcy, and HHcy worsened amyloid-induced barrier permeability. These effects were prevented by CAi and mCAi.
    CONCLUSION: Both HHcy and Aβ had detrimental effects on EC mitochondrial function and BBB permeability, and HHcy worsened the effects of Aβ. These effects were prevented by CAi and mCAi. The ability of mCAi to protect against Aβ and/or HHcy suggests a central role of the mitochondria in mediating vascular dysfunction in the presence of Aβ and/or HHcy. Our studies support the importance of better understanding mitochondrial and EC pathways responsible for cerebrovascular dysfunction in CAA and mixed dementias.
    DOI:  https://doi.org/10.1002/alz.054397
  29. Fundam Clin Pharmacol. 2022 Feb 03.
       BACKGROUND: Traumatic brain injury (TBI) is one of the most prevalent causes of permanent physical and cognitive disabilities. TBI pathology results from primary insults and a multi-mechanistic biochemical process, termed as secondary brain injury. Currently, there are no pharmacological agents for definitive treatment of patients with TBI.
    OBJECTIVES: This article is presented with the purpose of reviewing molecular mechanisms of TBI pathology, as well as potential strategies and agents against pathological pathways.
    METHODS: In this review article, materials were obtained by searching PubMed, Scopus, Elsevier, Web of Science, and Google Scholar. This search was considered without time limitation.
    RESULTS: Evidence indicates that oxidative stress and mitochondrial dysfunction are two key mediators of the secondary injury cascade in TBI pathology. TBI-induced oxidative damage results in the structural and functional impairments of cellular and subcellular components, such as mitochondria. Impairments of mitochondrial electron transfer chain and mitochondrial membrane potential result in a vicious cycle of free radical formation and cell apoptosis. The results of some preclinical and clinical studies, evaluating mitochondria-targeted therapies, such as mitochondria-targeted antioxidants and compounds with pleiotropic effects after TBI, are promising.
    CONCLUSIONS: As a proposed strategy in recent years, mitochondria-targeted multi-potential therapy is a new hope, waiting to be confirmed. Moreover, based on the available findings, biologics, such as stem cell-based therapy and transplantation of mitochondria are novel potential strategies for the treatment of TBI; however, more studies are needed to clearly confirm the safety and efficacy of these strategies.
    Keywords:  antioxidant; apoptosis; mitochondrion; oxidative stress; therapies; traumatic brain injury
    DOI:  https://doi.org/10.1111/fcp.12767
  30. Alzheimers Dement. 2021 Dec;17 Suppl 3 e050382
       BACKGROUND: Chronic high fat diet (HFD) consumption induces not only low-grade systemic inflammation and obese-insulin resistance, but also initiates neuroinflammation via activating myeloid differentiation factor 2 (MD-2)/toll-like receptor 4 (TLR4) complex, resulting in brain pathologies and cognitive impairment. Recently, MD-2 inhibitor (L6H21) has been shown to alleviate systemic inflammation. However, the effect of L6H21 on the cognitive function and brain pathologies of HFD-induced obese insulin-resistant condition has not been investigated.
    METHOD: Male Wistar rats were fed either a normal diet (ND; n=8) or HFD (n=40) for 16 weeks. After 12th week of dietary periods, HFD-fed rats were divided into 5 groups (n=8/group). Each group was orally administered either vehicle, 10 mg/kg/day of L6H21, 20 mg/kg/day of L6H21, 40 mg/kg/day of L6H21, or 300 mg/kg/day of metformin for 4 weeks. For ND-fed rats, animals were orally administered vehicle for 4 weeks. At the end of experimental protocol, the novel object location (NOL) test was examined in each animal. Then, blood was collected before animals were sacrificed. Brains were rapidly removed to determine microglial activity, dendritic spine density, inflammatory cytokines, mitochondrial function, and insulin signaling.
    RESULT: HFD-fed rats showed peripheral and brain insulin resistance, as indicated by impaired glucose tolerance and reduced brain insulin receptor substrate (IRS) level. In addition, HFD-fed rats developed brain pathologies, and cognitive impairment, as indicated by increased brain TNF-α level, increased microglial hyperactivity, increased mitochondrial reactive oxygen species (ROS), increased Bax/Bcl2 level, reduced dendritic spine density, and reduced preference indexes of NOL phase. Although L6H21 (20mg/kg/day, 40 mg/kg/day) and metformin alleviated HFD-induced brain pathologies and cognitive impairment, L6H21 at 40mg/kg/day and metformin had the highest efficacy for the neuroprotection in obese insulin-resistant rats through inhibiting neuroinflammation, oxidative stress, microglial activation, neuronal apoptosis and improving dendritic spine density, resulting in restoring cognitive function (p<0.05, Figure 1).
    CONCLUSION: These findings suggest that MD-2 inhibitor may be the other drug of choice for improving brain function in the obese-insulin resistant condition.
    DOI:  https://doi.org/10.1002/alz.050382
  31. Alzheimers Dement. 2021 Dec;17 Suppl 3 e053736
       BACKGROUND: Brain capillary endothelial cells (BCECs) are integral components of both the blood-brain barrier (BBB) and the neurovascular unit (NVU). Transport across the BBB is an important mediator of beta-amyloid (Aβ) accumulation in the brain and a contributing factor in the pathogenesis of Alzheimer's disease (AD). One of the receptors responsible for the transport of Aβ through the BBB is the low-density lipoprotein receptor-related protein 1 (LRP1). In addition, the development of AD and diabetes share many pathophysiological features including defective insulin signalling, impaired glucose metabolism, and cognitive decline. LRP1 is known to modulate insulin signalling by forming a dimer with insulin receptor beta when stimulated using insulin. Furthermore, LRP1 expression at the BBB is reduced during normal aging and in AD. Hence, we hypothesize that LRP1 activity in BCEC can be modulated using astaxanthin to improve Aβ clearance and insulin-mediated signalling at the BBB.
    METHOD: By using the established in vitro porcine brain capillary endothelial cell (pBCEC) model of the BBB, we analyzed the effects of astaxanthin on LRP1 expression, Aβ clearance, insulin-mediated signalling and other systemic dysfunctions associated with AD at the protein and mRNA level. We also examined the ultra-structures by electron microscopy.
    RESULT: pBCECs showed enhanced expression of LRP1 when treated with astaxanthin. We further observed improved insulin sensitivity when cells pre-incubated with astaxanthin were treated with Aβ1-40 , Aβ1-42 and Aβ1-40 + Aβ1-42 and insulin and further stimulated with insulin (10 nM). Increased expression of LRP1, Aβ-degrading enzymes, as well as autophagy and insulin signalling markers were observed when pBCECs pre-incubated with astaxanthin were further treated with amyloid beta peptides.
    CONCLUSION: Our results suggest that increased LRP1 expression by astaxanthin enhances insulin sensitivity, autophagy induction and improves Aβ degradation. Astaxanthin could thus be a promising therapeutic candidate for Alzheimer's disease.
    DOI:  https://doi.org/10.1002/alz.053736