Summary of Study ST003290

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR002042. The data can be accessed directly via it's Project DOI: 10.21228/M8NV6P This work is supported by NIH grant, U2C- DK119886.

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This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST003290
Study TitleHigh expression of oleoyl-ACP-hydrolase underpins life-threatening respiratory viral diseases
Study SummaryAlthough respiratory viral infections cause significant morbidity and mortality, it is unclear why some individuals succumb to severe disease. In patients hospitalized with avian A(H7N9) influenza, we investigated early drivers underpinning hypercytokinemia and fatal disease. Our transcriptomics studies identified differential expression of 10 early host genes, defined by 16 probe sets, between patients who recovered and died. Seven probe sets were specific for the same host gene encoding for a key enzyme mediating free fatty acid production, oleoyl-ACP-hydrolase (OLAH). High OLAH levels in fatal A(H7N9) patients were detected early after hospital admission and persisted until patients died. Conversely, patients who recovered had minimal OLAH expression throughout their hospital stay. High OLAH levels were also detected in patients hospitalized for severe infections with seasonal influenza virus, SARS-CoV-2, respiratory syncytial virus (RSV) and for multisystem inflammatory syndrome in children (MIS-C), while the main catalytic product of OLAH, oleic acid, was increased in hospitalized compared to ambulatory patients. Among healthy individuals and those with mild infections, however, OLAH was minimally detected. To understand how OLAH drives disease severity, we generated olah deficient mice. In contrast to wild-type mice, lethal influenza infection of olah-/- mice led to survival, milder disease, and markedly reduced lung viral loads, tissue damage, infection-driven pulmonary innate cell infiltration and inflammatory milieu. This phenotype was associated with differential lipid droplet dynamics, and reduced viral infection and inflammatory responses in macrophages. Supplementation of oleic acid, the main product of OLAH, increased influenza infection of macrophages and their inflammatory potential. Increased infectivity in the presence of olah was dependent on lipid droplet usage. Our findings define how expression of the key host enzyme, OLAH, drives life-threatening inflammation associated with respiratory viruses and propose OLAH as a potential early target for diagnosis and treatment of patients with severe disease. 
Institute
Peter Doherty Institute for Infection and Immunity
Last NameChua
First NameBrendon
Address792 Elizabeth St, Melbourne VIC 3000
Emailbychua@unimelb.edu.au
Phone+61383441130
Submit Date2024-06-26
Raw Data AvailableYes
Raw Data File Type(s)mzML
Analysis Type DetailLC-MS
Release Date2024-08-22
Release Version1
Brendon Chua Brendon Chua
https://dx.doi.org/10.21228/M8NV6P
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR002042
Project DOI:doi: 10.21228/M8NV6P
Project Title:High expression of oleoyl-ACP-hydrolase underpins life-threatening respiratory viral diseases
Project Summary:Although respiratory viral infections cause significant morbidity and mortality, it is unclear why some individuals succumb to severe disease. In patients hospitalized with avian A(H7N9) influenza, we investigated early drivers underpinning hypercytokinemia and fatal disease. Our transcriptomics studies identified differential expression of 10 early host genes, defined by 16 probe sets, between patients who recovered and died. Seven probe sets were specific for the same host gene encoding for a key enzyme mediating free fatty acid production, oleoyl-ACP-hydrolase (OLAH). High OLAH levels in fatal A(H7N9) patients were detected early after hospital admission and persisted until patients died. Conversely, patients who recovered had minimal OLAH expression throughout their hospital stay. High OLAH levels were also detected in patients hospitalized for severe infections with seasonal influenza virus, SARS-CoV-2, respiratory syncytial virus (RSV) and for multisystem inflammatory syndrome in children (MIS-C), while the main catalytic product of OLAH, oleic acid, was increased in hospitalized compared to ambulatory patients. Among healthy individuals and those with mild infections, however, OLAH was minimally detected. To understand how OLAH drives disease severity, we generated olah deficient mice. In contrast to wild-type mice, lethal influenza infection of olah-/- mice led to survival, milder disease, and markedly reduced lung viral loads, tissue damage, infection-driven pulmonary innate cell infiltration and inflammatory milieu. This phenotype was associated with differential lipid droplet dynamics, and reduced viral infection and inflammatory responses in macrophages. Supplementation of oleic acid, the main product of OLAH, increased influenza infection of macrophages and their inflammatory potential. Increased infectivity in the presence of olah was dependent on lipid droplet usage. Our findings define how expression of the key host enzyme, OLAH, drives life-threatening inflammation associated with respiratory viruses and propose OLAH as a potential early target for diagnosis and treatment of patients with severe disease. 
Institute:Peter Doherty Institute for Infection and Immunity
Last Name:Chua
First Name:Brendon
Address:792 Elizabeth St, Melbourne VIC 3000
Email:bychua@unimelb.edu.au
Phone:+61383441130

Subject:

Subject ID:SU003410
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Species Group:Mammals

Factors:

Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id local_sample_id Sample source Mouse Strain Infection status
SA356498231201_BC-7661_blank_4Extraction blank Extraction blank Extraction blank
SA356499231201_BC-7661_blank_3Extraction blank Extraction blank Extraction blank
SA356500231201_BC-7661_blank_2Extraction blank Extraction blank Extraction blank
SA356501231201_BC-7661_blank_1Extraction blank Extraction blank Extraction blank
SA356502231201_BC-7661_20Lung C57BL/6 wildtype Infected
SA356503231201_BC-7661_19Lung C57BL/6 wildtype Infected
SA356504231201_BC-7661_18Lung C57BL/6 wildtype Infected
SA356505231201_BC-7661_17Lung C57BL/6 wildtype Infected
SA356506231201_BC-7661_16Lung C57BL/6 wildtype Infected
SA356507231201_BC-7661_14Lung C57BL/6 wildtype Naïve
SA356508231201_BC-7661_15Lung C57BL/6 wildtype Naïve
SA356509231201_BC-7661_13Lung C57BL/6 wildtype Naïve
SA356510231201_BC-7661_12Lung C57BL/6 wildtype Naïve
SA356511231201_BC-7661_11Lung C57BL/6 wildtype Naïve
SA356512231201_BC-7661_10Lung olah-/- Infected
SA356513231201_BC-7661_9Lung olah-/- Infected
SA356514231201_BC-7661_8Lung olah-/- Infected
SA356515231201_BC-7661_7Lung olah-/- Infected
SA356516231201_BC-7661_6Lung olah-/- Infected
SA356517231201_BC-7661_2Lung olah-/- Naïve
SA356518231201_BC-7661_5Lung olah-/- Naïve
SA356519231201_BC-7661_4Lung olah-/- Naïve
SA356520231201_BC-7661_3Lung olah-/- Naïve
SA356521231201_BC-7661_1Lung olah-/- Naïve
Showing results 1 to 24 of 24

Collection:

Collection ID:CO003403
Collection Summary:Whole lungs were harvested, weighed, minced and freeze dried. Prior to lipid extraction, freeze dried lungs were incubated with 200µl of ice-cold 60% methanol containing 0.01% (w/v) butylated hydroxytoluene (BHT) and incubated overnight at -20°C
Sample Type:Lung

Treatment:

Treatment ID:TR003419
Treatment Summary:Mice (olah-/- or C57BL/6) were bred and maintained in the Biological Research Facility in the Department of Microbiology and Immunology at The University of Melbourne. Following weaning (~3 weeks old), mice were placed on a normal maintenance diet (Barastoc, Ridley Corporation, Australia). Animal experiments were conducted in accordance with the Australian National Health and Medical Research Council Code of Practice for the Care and Use of Animals for Scientific Purposes Guidelines and institutional regulations following approval (permit numbers 1714304, 1614073 and 20532) by The University of Melbourne Animal Ethics Committee. Influenza virus infection was performed under light anaesthesia with isofluorane and intranasal instillation (30µl) with 2x104 plaque forming units of A/HK/x31 (X31; H3N2). One day following infection, lungs were harvested, weighed and snap frozen in preparation for lipid extraction. Naïve mice were treated as above except that influenza virus infection was not performed. 

Sample Preparation:

Sampleprep ID:SP003417
Sampleprep Summary:Whole lungs were harvested, weighed, minced and freeze dried. Prior to lipid extraction, freeze dried lungs were incubated with 200µl of ice-cold 60% methanol containing 0.01% (w/v) butylated hydroxytoluene (BHT) and incubated overnight at -20°C. SPLASH® LIPIDOMIX® Mass Spec Standard (10µl; Cat. 330707, Avanti Polar Lipids, Birmingham, AL, USA) and deuterated saturated/monounsaturated fatty acid standards (10µl; Cayman Chemical, USA) were added to each sample and tissue homogenised with a Bullet Blender (Next Advance, Troy, NY, USA) and sonicated for 20 minutes. Monophasic lipid extraction was performed according to Lydic et al (Lydic et al., 2015, DOI: 10.1016/j.ymeth.2015.04.014) wherein 120 μl of water, 420 μl of methanol with 0.01% (w/v) BHT, and 270 μl of chloroform were added to all samples prior to thorough vortexing and incubated on a shaker at 1400 rpm for 30 mins. Following centrifugation, supernatants containing lipids were transferred to new tubes and the remaining pellets re-extracted by adding 100µl of water and 400µl of chloroform:methanol (1:2 v/v) containing 0.01% (w/v) BHT and incubation on a shaker at 1000 rpm for 15 mins. Supernatants from each extraction round were pooled, dried by evaporation under vacuum and freeze-dried. Lyophilised samples were then resuspended in chloroform:methanol (1:9 v/v) containing 0.01% BHT and centrifuged to remove precipitates.

Combined analysis:

Analysis ID AN005388 AN005389
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Thermo Vanquish Thermo Vanquish
Column Waters ACQUITY UPLC HSS T3 (150 x 1mm,1.8um) Waters ACQUITY UPLC HSS T3 (150 x 1mm,1.8um)
MS Type ESI ESI
MS instrument type Orbitrap Orbitrap
MS instrument name Thermo Fusion Tribrid Orbitrap Thermo Fusion Tribrid Orbitrap
Ion Mode POSITIVE NEGATIVE
Units pmol/mg lung tissue pmol/mg lung tissue

Chromatography:

Chromatography ID:CH004085
Chromatography Summary:Solvent A: 6/4 (v/v) acetonitrile/water with 5 uM medronic acid and 10 mM ammonium acetate; Solvent B: 9/1 (v/v) isopropanol/acetonitrile with 10 mM ammonium acetate
Instrument Name:Thermo Vanquish
Column Name:Waters ACQUITY UPLC HSS T3 (150 x 1mm,1.8um)
Column Temperature:50
Flow Gradient:[Time(min), %B solvent]: [0,3], [5,3], [10,70], [26,99], [29,99], [29.1,3], [33,3] 
Flow Rate:60 uL/min
Solvent A:60% acetonitrile/40% water; 5 uM medronic acid; 10 mM ammonium acetate
Solvent B:90% isopropyl alcohol/10% acetonitrile; 10 mM ammonium acetate
Chromatography Type:Reversed phase

MS:

MS ID:MS005115
Analysis ID:AN005388
Instrument Name:Thermo Fusion Tribrid Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:Top speed data-dependent scan with a cycle time of 1s was used. Within each cycle, a full-scan MS-spectra were acquired firstly in the Orbitrap at a mass resolving power of 120,000 across an m/z range of 300–2000 using quadrupole isolation, followed by higher-energy collisional dissociation (HCD)-MS/MS at a mass resolving power of 15,000 (at m/z 200) and a normalized collision energy (NCE) of 27% at positive mode and 30% in negative mode with an m/z isolation window of 1.
Ion Mode:POSITIVE
  
MS ID:MS005116
Analysis ID:AN005389
Instrument Name:Thermo Fusion Tribrid Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:Top speed data-dependent scan with a cycle time of 1s was used. Within each cycle, a full-scan MS-spectra were acquired firstly in the Orbitrap at a mass resolving power of 120,000 across an m/z range of 300–2000 using quadrupole isolation, followed by higher-energy collisional dissociation (HCD)-MS/MS at a mass resolving power of 15,000 (at m/z 200) and a normalized collision energy (NCE) of 27% at positive mode and 30% in negative mode with an m/z isolation window of 1.
Ion Mode:NEGATIVE
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