DOI: 10.5061/dryad.dbrv15f9j
title: Restoring hippocampal glucose metabolism rescues cognition across Alzheimer disease pathologies

description: Impaired cerebral glucose metabolism is a pathologic feature of Alzheimer
 Disease (AD) and recent proteomic studies highlight a disruption of glial
 carbohydrate metabolism with disease progression.  Here, we
 report that inhibition of indoleamine-2,3-dioxygenase 1 (IDO1), which
 metabolizes tryptophan to kynurenine (KYN) in the first step of the
 kynurenine pathway, rescues hippocampal memory function and plasticity in
 preclinical models of amyloid and tau pathology by restoring astrocytic
 metabolic support of neurons.  Activation of IDO1 in astrocytes
 by amyloid-beta and tau oligomers, two major pathologic effectors in AD,
 increases KYN and suppresses glycolysis in an AhR-dependent manner.
 Conversely, pharmacological IDO1 inhibition restores glycolysis and
 lactate production. In amyloid-producing APPSwe-PS1∆E9 and 5XFAD mice and
 in tau-producing P301S mice, IDO1 inhibition restores spatial memory and
 improves hippocampal glucose metabolism by metabolomic and MALDI-MS
 analyses. IDO1 blockade also rescues hippocampal long-term potentiation in
 a monocarboxylate transporter (MCT)-dependent manner, suggesting that IDO1
 activity disrupts astrocytic metabolic support of neurons. Indeed, in
 vitro mass-labeling of human astrocytes demonstrates that IDO1 regulates
 astrocyte generation of lactate that is then taken up by human
 neurons.  In co-cultures of astrocytes and neurons derived from
 AD subjects, deficient astrocyte lactate production and transfer to
 neurons was corrected by IDO1 inhibition, resulting in improved neuronal
 glucose metabolism.  Thus, IDO1 activity disrupts astrocytic
 metabolic support of neurons across both amyloid and tau pathologies and
 in a model of AD iPSC-derived neurons. These findings also suggest that
 IDO1 inhibitors developed for adjunctive therapy in cancer could be
 repurposed for treatment of amyloid- and tau-mediated neurodegenerative
 diseases.
description: Metabolites were extracted from isolated astrocytes and neurons
 in a -80˚C 80:20 methanol:water solution in a volume of 1.5 ml per 1x106
 cells, vortexed, incubated on dry ice for 10 min, and centrifuged at
 16,000 g for 20 min, and the supernatant was assayed by LC-MS analysis.
 Extracts were analyzed within 24 hr by liquid chromatography coupled to a
 mass spectrometer (LC-MS). The LC-MS method involved hydrophilic
 interaction chromatography (HILIC) coupled to the Q Exactive PLUS mass
 spectrometer (Thermo Scientific) 7 (17). The LC separation was performed
 on an XBridge BEH Amide column (150 mm 3 2.1 mm, 2.5 mm particle size,
 Waters, Milford, MA). Solvent A was 95%: 5% H<sub>2</sub>O:
 acetonitrile with 20 mM ammonium bicarbonate, and solvent B was
 acetonitrile. The gradient was 0 min, 85% B; 2 min, 85% B; 3 min, 80% B; 5
 min, 80% B; 6 min, 75% B; 7 min, 75% B; 8 min, 70% B; 9 min, 70% B; 10
 min, 50% B; 12 min, 50% B; 13 min, 25% B; 16 min, 25% B; 18 min, 0% B; 23
 min, 0% B; 24 min, 85% B; and 30 min, 85% B. Other LC parameters are: flow
 rate 150 ml/min, column temperature 25°C, and injection volume 10 mL and
 autosampler temperature was 5°C. The mass spectrometer was operated in
 both negative and positive ion mode for the detection of metabolites.
 Other MS parameters were: resolution of 140,000 at m/z 200, automatic gain
 control (AGC) target at 3e6, maximum injection time of 30 ms and scan
 range of m/z 75- 1000. Raw LC/MS data were converted to mzXML format using
 the command line “msconvert” utility (18). Data were obtained with MAVEN
 software (19, 20). For identification of hexose phosphates and glycolytic
 intermediates, capillary electrophoresis mass spectroscopy was used as
 previously described (21). In brief, adherent cells on dishes were washed
 with 5% mannitol aqueous solution at room temperature. The cells were
 immersed in 400 μl methanol for 30 s, and 275 μl of the Internal Standard
 Solution (10 μM, Solution ID: H3304-1002, Human Metabolome Technologies)
 for 30 s. The extraction liquid was centrifuged at 2,300g for 5 min at 4
 °C. The supernatant (400 μl) was centrifugally filtered at 9,100g for 4 h
 at 4°C through a 5-kDa cut-off filter (Millipore) to remove proteins, and
 then the filtrate was lyophilized and suspended in 25 μl Milli-Q water.
 The metabolite suspension was analysed by capillary electrophoresis time
 of flight mass spectrometry (CE-TOF/MS) using an Agilent capillary
 electrophoresis (CE) system equipped with an Agilent 6210 TOFMS, an 1100
 isocratic high-performance
description: # Restoring hippocampal glucose metabolism rescues cognition across
 Alzheimer disease pathologies Access this dataset on Dryad](DOI:
 10.5061/dryad.dbrv15f9j) The excel files located in this dryad dataset
 contain normalized values (z-score) of multi-analyte cytokine array
 (Luminex), qRT-PCR of selected genes, as well as metabolomics from several
 different experiments. Experiments are from both in vitro and in vivo
 astrocytes as well as hippocampus tissue. Experimental conditions are
 listed below.  ## Description of Data and file structure Source Data
 1H.xlsx qRT-PCR of AhR-signaling genes in vitro post-natal Mouse
 Astrocytes treated with Vehicle (Veh), oTau+oAB (oligomeric Tau +
 oligomeric Amyloid beta), Ido1 inhibitor PF068. First column contains Gene
 names; subsequent columns contain control and experimental groups.  Source
 Data 1I.xlsx Hif-1a signaling qRT-PCR in vitro post-natal Mouse Astrocytes
 treated with Vehicle (Veh), oTau+oAB (oligomeric Tau + oligomeric Amyloid
 beta), Ido1 inhibitor PF068. First column contains Gene names; subsequent
 columns contain control and experimental groups Source Data 1J.xlsx
 Glycolytic metabolites in vitro astrocytes treated with oTau+oAB +/-
 PF068. First column contain metabolites; subsequent columns contain
 control and experimental groups.  Source Data 3M.xlsx qRT-PCR of AhR
 signaling genes from isolated hippocampi of 5xFAD mice +/- PF068
 treatment. First column contains Gene names; subsequent columns contain
 control and experimental groups. Source Data 3N.xlsx qRT-PCR of Hif1a
 signaling genes from isolated hippocampi of 5xFAD mice +/- PF068
 treatment. First column contains Gene names; subsequent columns contain
 control and experimental groups.  Source Data 4F_AhR.xlsx qRT-PCR of AhR
 signaling genes from isolated hippocampi of PS19 mice +/- PF068 treatment.
 First column contains Gene names; subsequent columns contain control and
 experimental groups.  Source Data 4F_Hif1a.xlsx qRT-PCR of Hif1a signaling
 genes from isolated hippocampi of PS19 mice +/- PF068 treatment. First
 column contains Gene names; subsequent columns contain control and
 experimental groups.  Source Data 5C.xlsx 13 common metabolites
 significantly altered among 3 different AD model mice treated with PF068.
 LC/MS metabolomics of the thirteen metabolites isolated hippocampi from
 5xFAD mice +/- PF068 treatment. First column contains metabolite names;
 subsequent columns contain control and experimental groups.  Source Data
 5F.xlsx 13 common metabolites significantly altered among 3 different AD
 model mice treated with PF068. LC/MS metabolomics of the thirteen
 metabolites isolated hippocampi from APP/PS1 mice +/- PF068 treatment.
 First column contains metabolite names; subsequent columns contain control
 and experimental groups.  Source Data 5G.xlsx 13 common metabolites
 significantly altered among 3 different AD model mice treated with PF068.
 LC/MS metabolomics of the thirteen metabolites isolated hippocampi from
 APP/PS1xIDO1-/- mice +/- PF068 treatment. First column contains metabolite
 names; subsequent columns contain control and experimental groups.  Source
 Data 5H.xlsx 13 common metabolites significantly altered among 3 different
 AD model mice treated with PF068. LC/MS metabolomics of the thirteen
 metabolites isolated hippocampi from PS19 mice +/- PF068 treatment. First
 column contains metabolite names; subsequent columns contain control and
 experimental groups.  Source Data Supplementary Figure 3I.xlsx
 Multi-analyte cytokine assay results of in vitro mouse astrocyte treated
 with veh, PF068, TNFa+C1q+IL1a +/- PF068. First column contains cytokine
 analytes; tube Source Data Supplementary Figure 8A.xlsx Untargeted
 metabolomics significant q&lt;0.05 metabolites PF068 vs. Veh from isolated
 hippocampi of APP/PS1 mice +/- PF068.  Source Data Supplementary Figure
 8B.xlsx Untargeted metabolomics significant q&lt;0.05 metabolites PF068
 vs. Veh from isolated hippocampi of 5xFAD mice +/- PF068.  Source Data
 Supplementary Figure 8C.xlsx Untargeted metabolomics significant q&lt;0.05
 metabolites PF068 vs. Veh from isolated hippocampi of PS19 mice +/-
 PF068.