Summary of Study ST002117

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 PR001342. The data can be accessed directly via it's Project DOI: 10.21228/M87X3H 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 IDST002117
Study TitleMicrobial Toll/interleukin-1 domain NADase activities
Study SummaryRecent work indicates that Toll/Interleukin-1 receptor (TIR) domain-containing proteins degrade NAD. Given increasing attention to how the gut microbiome contributes to NAD metabolism and healthy growth, we examined the representation and expressed NADase activities of its TIR domains. The NADase activities of 151 bacterial TIRs were characterized in vitro. Gut bacterial strains representing the diversity of TIRs observed in the microbiome, and activities observed in vitro, were introduced into germ-free mice fed defined diets. Mass spectrometry of cecal metabolites disclosed that a product of TIR NADase activity, variant cyclic-ADPR-x, distinguished colonized from germ-free animals. Mass spectrometry and microbial RNA-Seq of gnotobiotic mice colonized with one, two and all members of this bacterial consortium, identified Bacteroides xylanisolvens as the principal in vivo source of v-cADPR-x. Guided by bioinformatic analyses of biochemically validated TIR domains, we determined that compared to age-matched healthy Bangladeshi children, those with acute malnutrition had significantly lower fecal levels of TIRs known or predicted to generate v-cADPR-x, and of this metabolite. These results indicate that v-cADPR-x may be an informative biomarker of healthy gut microbiome development.
Institute
Washington University in St. Louis
Last NameSasaki
First NameYo
Address4515 McKinley Ave St.Louis MO
Emailsasaki@wustl.edu
Phone314-362-4653
Submit Date2022-03-22
Raw Data AvailableYes
Raw Data File Type(s)d
Analysis Type DetailLC-MS
Release Date2022-03-30
Release Version1
Yo Sasaki Yo Sasaki
https://dx.doi.org/10.21228/M87X3H
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR001342
Project DOI:doi: 10.21228/M87X3H
Project Title:Microbial Toll/interleukin-1 domain metabolite study.
Project Summary:The Characterization of NADase activities of 151 bacterial TIRs.
Institute:Washington University in St. Louis
Last Name:Yo
First Name:Sasaki
Address:4515 McKinley Ave St.Louis MO
Email:sasaki@wustl.edu
Phone:314-362-4653

Subject:

Subject ID:SU002202
Subject Type:Bacteria
Subject Species:Bacteroides xylanisolvens

Factors:

Subject type: Bacteria; Subject species: Bacteroides xylanisolvens (Factor headings shown in green)

mb_sample_id local_sample_id Cage Microbiota Diet Age (months) WLZ Health phenotype
SA203044m06c1 26-member NAD-precursor deficient NA NA NA
SA203045m02d1 26-member NAD-precursor deficient NA NA NA
SA203046m18b1 26-member NAD-precursor deficient NA NA NA
SA203047m15a-S5A1 26-member NAD-precursor sufficient NA NA NA
SA203048m13a-S5A1 26-member NAD-precursor sufficient NA NA NA
SA203049m07a-S5A1 26-member NAD-precursor sufficient NA NA NA
SA203050m11a-S5A1 26-member NAD-precursor sufficient NA NA NA
SA203051m11a1 BthetaBxylan NAD-precursor deficient NA NA NA
SA203052m17b1 BthetaBxylan NAD-precursor deficient NA NA NA
SA203053m14b1 BthetaBxylan NAD-precursor deficient NA NA NA
SA203054m14a1 Btheta NAD-precursor deficient NA NA NA
SA203055m05b1 Btheta NAD-precursor deficient NA NA NA
SA203056m13a1 Btheta NAD-precursor deficient NA NA NA
SA203057m01c1 Bxylan NAD-precursor deficient NA NA NA
SA203058m03c1 Bxylan NAD-precursor deficient NA NA NA
SA203059m07a1 Bxylan NAD-precursor deficient NA NA NA
SA203060m02a1 Germfree NAD-precursor deficient NA NA NA
SA203061m08a1 Germfree NAD-precursor deficient NA NA NA
SA203062m01a1 Germfree NAD-precursor deficient NA NA NA
SA203063m01b-S5A1 Germfree NAD-precursor sufficient NA NA NA
SA203064m07b-S5A1 Germfree NAD-precursor sufficient NA NA NA
SA203065m09a-S5A1 Germfree NAD-precursor sufficient NA NA NA
SA203066m08a-S5A1 Germfree NAD-precursor sufficient NA NA NA
SA203067m07c2 26-member NAD-precursor deficient NA NA NA
SA203068m01d2 26-member NAD-precursor deficient NA NA NA
SA203069m11b2 26-member NAD-precursor deficient NA NA NA
SA203070m01a-S5A2 26-member NAD-precursor sufficient NA NA NA
SA203071m06b-S5A2 26-member NAD-precursor sufficient NA NA NA
SA203072m18a-S5A2 26-member NAD-precursor sufficient NA NA NA
SA203073m10b-S5A2 26-member NAD-precursor sufficient NA NA NA
SA203074m15b2 BthetaBxylan NAD-precursor deficient NA NA NA
SA203075m16b2 BthetaBxylan NAD-precursor deficient NA NA NA
SA203076m03d2 BthetaBxylan NAD-precursor deficient NA NA NA
SA203077m12a2 Btheta NAD-precursor deficient NA NA NA
SA203078m04c2 Btheta NAD-precursor deficient NA NA NA
SA203079m15a2 Btheta NAD-precursor deficient NA NA NA
SA203080m08c2 Bxylan NAD-precursor deficient NA NA NA
SA203081m04d2 Bxylan NAD-precursor deficient NA NA NA
SA203082m05c2 Bxylan NAD-precursor deficient NA NA NA
SA203083m05a2 Germfree NAD-precursor deficient NA NA NA
SA203084m04a2 Germfree NAD-precursor deficient NA NA NA
SA203085m03a2 Germfree NAD-precursor deficient NA NA NA
SA203086m11b-S5A2 Germfree NAD-precursor sufficient NA NA NA
SA203087m14a-S5A2 Germfree NAD-precursor sufficient NA NA NA
SA203088m13b-S5A2 Germfree NAD-precursor sufficient NA NA NA
SA203089m17a-S5A2 Germfree NAD-precursor sufficient NA NA NA
SA203090m04b-S5A3 26-member NAD-precursor deficient NA NA NA
SA203091m06a-S5A3 26-member NAD-precursor deficient NA NA NA
SA203092m12b-S5A3 26-member NAD-precursor deficient NA NA NA
SA203093m10a-S5A3 26-member NAD-precursor deficient NA NA NA
SA203094m10a3 Btheta NA-supplementeded NA NA NA
SA203095m02c3 Btheta NA-supplementeded NA NA NA
SA203096m03b3 Btheta NA-supplementeded NA NA NA
SA203097m13b3 Bxylan NA-supplementeded NA NA NA
SA203098m09b3 Bxylan NA-supplementeded NA NA NA
SA203099m10b3 Bxylan NA-supplementeded NA NA NA
SA203103m16a-S5A3 Germfree NAD-precursor deficient NA NA NA
SA203104m05b-S5A3 Germfree NAD-precursor deficient NA NA NA
SA203105m02a-S5A3 Germfree NAD-precursor deficient NA NA NA
SA203106m14b-S5A3 Germfree NAD-precursor deficient NA NA NA
SA203100m06a3 Germfree NA-supplementeded NA NA NA
SA203101m09a3 Germfree NA-supplementeded NA NA NA
SA203102m04b3 Germfree NA-supplementeded NA NA NA
SA203107m04a-S5A4 26-member NAD-precursor deficient NA NA NA
SA203108m09b-S5A4 26-member NAD-precursor deficient NA NA NA
SA203109m12a-S5A4 26-member NAD-precursor deficient NA NA NA
SA203110m03a-S5A4 26-member NAD-precursor deficient NA NA NA
SA203111m18a4 Btheta NA-supplementeded NA NA NA
SA203112m16a4 Btheta NA-supplementeded NA NA NA
SA203113m17a4 Btheta NA-supplementeded NA NA NA
SA203114m07b4 Bxylan NA-supplementeded NA NA NA
SA203115m12b4 Bxylan NA-supplementeded NA NA NA
SA203116m06b4 Bxylan NA-supplementeded NA NA NA
SA203120m02b-S5A4 Germfree NAD-precursor deficient NA NA NA
SA203121m05a-S5A4 Germfree NAD-precursor deficient NA NA NA
SA203122m03b-S5A4 Germfree NAD-precursor deficient NA NA NA
SA203123m08b-S5A4 Germfree NAD-precursor deficient NA NA NA
SA203117m08b4 Germfree NA-supplementeded NA NA NA
SA203118m02b4 Germfree NA-supplementeded NA NA NA
SA203119m01b4 Germfree NA-supplementeded NA NA NA
SA203124PS.079NA NA NA 10.3 -2.7 Malnourished
SA203125PS.057-10.4NA NA NA 10.4 -3.8 Malnourished
SA203126PS.057-10.6NA NA NA 10.6 -2.81 Malnourished
SA203127BG1C0473-11NA NA NA 11 -0.82 Healthy
SA203128PS.158NA NA NA 12.5 -3.48 Malnourished
SA203129BG1C0466-12NA NA NA 12 0.2 Healthy
SA203130PS.120-13.3NA NA NA 13.3 -3.16 Malnourished
SA203131PS.120-13.9NA NA NA 13.9 -2.17 Malnourished
SA203132BG1C0466-13NA NA NA 13 -0.3 Healthy
SA203133BG1C0473-13NA NA NA 13 -0.93 Healthy
SA203134PS.132NA NA NA 14.1 -3.01 Malnourished
SA203135Bgsng8064NA NA NA 15 0.6 Healthy
SA203136PS.170NA NA NA 16.2 -3.5 Malnourished
SA203137PS.004NA NA NA 21.9 -2.03 Malnourished
SA203138Bgsng8119NA NA NA 25 1.7 Healthy
SA203139Bgsng8169-28NA NA NA 28 - Healthy
SA203140Bgsng8169-30NA NA NA 30 1.77 Healthy
SA203141Bgsng8153NA NA NA 6 1.58 Healthy
SA203142BG1C0462NA NA NA 8 -0.43 Healthy
SA203143PS.011NA NA NA 9.9 -5.54 Malnourished
Showing results 1 to 100 of 100

Collection:

Collection ID:CO002195
Collection Summary:Cecal contents were collected on 10 uL plastic inoculation loops at the time of sacrifice, snap frozen in liquid nitrogen, and stored at -80 °C. Aliquots were kept frozen, removed from the loops and weighed.
Sample Type:Cecum
Storage Conditions:-80℃

Treatment:

Treatment ID:TR002214
Treatment Summary:Germ-free C57BL/6J mice were reared in plastic flexible film gnotobiotic isolators (Class Biologically Clean) at 23 °C under a strict 12-hour light cycle (lights on at 0600h, off at 1800h). Animals were weaned onto an autoclaved, low-fat, plant polysaccharide-rich chow (catalog number 2018S, Envigo) that was administered ad libitum. Five days prior to colonization, 8-week-old male mice were switched to the NAD precursor-sufficient diet; 2 days later, they were either placed on a NAD precursor-deficient diet, or a NA-supplemented diet or were continued on the NAD precursor-sufficient diet.

Sample Preparation:

Sampleprep ID:SP002208
Sampleprep Summary:10 uL of ice-cold methanol was added per mg of sample, which was then thawed on ice. The mixture was subsequently shaken for two minutes in a BioSpec bead beater. Samples were then centrifuged for 10 minutes at 3,000 x g at 4 °C; a 150 uL aliquot of the supernatant was transferred to a new tube containing 150 uL ice-cold chloroform, then mixed by pulse vortexing and centrifuged for 10 minutes at 3,000 x g at 4 °C. 100 uL of the methanol fraction was transferred to a new tube containing 100 uL of ice-cold chloroform, mixed by pulse-vortexing, and centrifuged for 10 minutes at 3,000 x g at 4 °C. 70 uL was transferred to a new tube and lyophilized in a Labconco SpeedVac.

Combined analysis:

Analysis ID AN003465
Analysis type MS
Chromatography type Reversed phase
Chromatography system Agilent 1290 Infinity
Column Waters Atlantis C18
MS Type ESI
MS instrument type Triple quadrupole
MS instrument name Agilent 6490 QQQ
Ion Mode POSITIVE
Units pmol/mg

Chromatography:

Chromatography ID:CH002559
Instrument Name:Agilent 1290 Infinity
Column Name:Waters Atlantis C18
Chromatography Type:Reversed phase

MS:

MS ID:MS003226
Analysis ID:AN003465
Instrument Name:Agilent 6490 QQQ
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:Metabolites were quantified by MassHunter quantitative analysis tool (Agilent)
Ion Mode:POSITIVE
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