Summary of Study ST001667

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 PR001037. The data can be accessed directly via it's Project DOI: 10.21228/M8NM4H This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

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 IDST001667
Study TitleLC-MS metabolomics analysis of ricin-induced and fasting hypoglycemia (part-II)
Study SummaryMice were subjected to ricin exposure or fasting conditions for 2 hours, 8 hours, or an overnight period. Following treatment, livers were removed and metabolites were extracted and analyzed by LC-MS.
Institute
Montana State University
Last NameKempa
First NameJake
Address103 Chemistry and Biochemistry Building, Bozeman, Montana, 59717, USA
Emailjkempa97@gmail.com
Phone4067997200
Submit Date2021-01-19
Raw Data AvailableYes
Raw Data File Type(s)mzML
Analysis Type DetailLC-MS
Release Date2022-11-29
Release Version1
Jake Kempa Jake Kempa
https://dx.doi.org/10.21228/M8NM4H
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR001037
Project DOI:doi: 10.21228/M8NM4H
Project Title:NMR and MS Metabolomics Reveal a Distinct Metabolic State of Ricin-induced Hypoglycemia
Project Type:Targeted NMR and Untargeted MS
Project Summary:Ricin toxin is a ribosome inactivating protein. Due to its toxic and chemical properties, ricin is a potential agent of bioterrorism but has also been studied for therapeutic use in immunotoxins. Previous research by our group has demonstrated lethal hypoglycemia associated with ricin toxicity. Research efforts have focused on better understanding this ricin-induced metabolic state of hypoglycemia to aid in better understanding the systemic effects of hypoglycemia, and the use of ricin in immunotherapy. Here, we have used a mouse model to characterize liver metabolome changes associated with hypoglycemia induced by two conditions. Mice were challenged with intraperitoneal injections of ricin at high and low doses for 2 hrs, 8 hrs, and overnight. To understand how ricin-induced hypoglycemia differs from that of fasting, another group of mice had food withheld for 8-hours and overnight. 1H NMR-based metabolomics was performed on polar molecules extracted from mouse livers, with metabolites annotated using Chenomx software. MetaboAnalyst was employed for multivariate statistical analysis. In this study, similar decreases in blood glucose in mice were observed following injection of a lethal dose of ricin and with overnight fasting. NMR analyses identified 59 polar metabolites present in mice livers from all treatments. Multivariate statistical analyses were used to evaluate global metabolic state differences. Results from these analyses indicated that the profiled liver metabolomes for mice subjected to the two conditions differ significantly at both 8 hours and overnight. Additionally, ricin treatment with a lethal dose reveal a progression of metabolic changes over time, from 2 to 22 hours. Additionally, mass spectrometry supported these findings, and NMR analyses revealed key metabolites that contribute to these differences. While both ricin and fasting induce hypoglycemia, the metabolic states resulting from these two conditions are different. Further analyses may give insights into mechanisms of ricin toxicity, specific metabolic pathways that are altered, and potential treatments for hypoglycemia. We propose to extend these studies to insulin-induced hypoglycemia.
Institute:Montana State University
Department:Chemistry & Biochemistry
Laboratory:Copié
Last Name:Kempa
First Name:Jake
Address:103 Chemistry and Biochemistry Building, Bozeman, Montana, 59717, USA
Email:jkempa97@gmail.com
Phone:4067997200

Subject:

Subject ID:SU001744
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090

Factors:

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

mb_sample_id local_sample_id Treatment
SA1522431--13Control | Timepoint:Overnight
SA1522441--12Control | Timepoint:Overnight
SA1522451--14Control | Timepoint:Overnight
SA1522461--16Control | Timepoint:Overnight
SA1522471--1Control | Timepoint:Overnight
SA1522481--10Control | Timepoint:Overnight
SA1522491--15Control | Timepoint:Overnight
SA1522501--11Control | Timepoint:Overnight
SA1522511--4Control | Timepoint:Overnight
SA1522521--3Control | Timepoint:Overnight
SA1522531--2Control | Timepoint:Overnight
SA1522541--9Control | Timepoint:Overnight
SA1522551--5Control | Timepoint:Overnight
SA1522561--6Control | Timepoint:Overnight
SA1522571--8Control | Timepoint:Overnight
SA1522581--7Control | Timepoint:Overnight
SA1522597--8Fast | Timepoint:8h
SA1522607--9Fast | Timepoint:8h
SA1522617--7Fast | Timepoint:8h
SA1522627--10Fast | Timepoint:8h
SA1522637--1Fast | Timepoint:8h
SA1522647--6Fast | Timepoint:8h
SA1522657--2Fast | Timepoint:8h
SA1522667--3Fast | Timepoint:8h
SA1522677--4Fast | Timepoint:8h
SA1522687--5Fast | Timepoint:8h
SA1522692--11Fast | Timepoint:Overnight
SA1522702--10Fast | Timepoint:Overnight
SA1522712--12Fast | Timepoint:Overnight
SA1522722--9Fast | Timepoint:Overnight
SA1522732--16Fast | Timepoint:Overnight
SA1522742--15Fast | Timepoint:Overnight
SA1522752--13Fast | Timepoint:Overnight
SA1522762--14Fast | Timepoint:Overnight
SA1522772--3Fast | Timepoint:Overnight
SA1522782--1Fast | Timepoint:Overnight
SA1522792--8Fast | Timepoint:Overnight
SA1522802--2Fast | Timepoint:Overnight
SA1522812--4Fast | Timepoint:Overnight
SA1522822--7Fast | Timepoint:Overnight
SA1522832--6Fast | Timepoint:Overnight
SA1522842--5Fast | Timepoint:Overnight
SA1522855--7High Dose Ricin | Timepoint:2h
SA1522865--8High Dose Ricin | Timepoint:2h
SA1522875--6High Dose Ricin | Timepoint:2h
SA1522885--2High Dose Ricin | Timepoint:2h
SA1522895--1High Dose Ricin | Timepoint:2h
SA1522905--10High Dose Ricin | Timepoint:2h
SA1522915--9High Dose Ricin | Timepoint:2h
SA1522925--3High Dose Ricin | Timepoint:2h
SA1522935--4High Dose Ricin | Timepoint:2h
SA1522945--5High Dose Ricin | Timepoint:2h
SA1522959--2High Dose Ricin | Timepoint:8h
SA1522969--1High Dose Ricin | Timepoint:8h
SA1522979--10High Dose Ricin | Timepoint:8h
SA1522989--8High Dose Ricin | Timepoint:8h
SA1522999--9High Dose Ricin | Timepoint:8h
SA1523009--6High Dose Ricin | Timepoint:8h
SA1523019--7High Dose Ricin | Timepoint:8h
SA1523029--5High Dose Ricin | Timepoint:8h
SA1523039--3High Dose Ricin | Timepoint:8h
SA1523049--4High Dose Ricin | Timepoint:8h
SA1523056--5High Dose Ricin | Timepoint:Overnight
SA1523066--4High Dose Ricin | Timepoint:Overnight
SA1523076--6High Dose Ricin | Timepoint:Overnight
SA1523086--3High Dose Ricin | Timepoint:Overnight
SA1523096--17High Dose Ricin | Timepoint:Overnight
SA1523106--16High Dose Ricin | Timepoint:Overnight
SA1523116--15High Dose Ricin | Timepoint:Overnight
SA1523126--18High Dose Ricin | Timepoint:Overnight
SA1523136--19High Dose Ricin | Timepoint:Overnight
SA1523146--20High Dose Ricin | Timepoint:Overnight
SA1523156--14High Dose Ricin | Timepoint:Overnight
SA1523166--13High Dose Ricin | Timepoint:Overnight
SA1523176--8High Dose Ricin | Timepoint:Overnight
SA1523186--10High Dose Ricin | Timepoint:Overnight
SA1523196--11High Dose Ricin | Timepoint:Overnight
SA1523206--12High Dose Ricin | Timepoint:Overnight
SA1523216--7High Dose Ricin | Timepoint:Overnight
SA1523223--3Low Dose Ricin | Timepoint:2h
SA1523233--4Low Dose Ricin | Timepoint:2h
SA1523243--2Low Dose Ricin | Timepoint:2h
SA1523253--1Low Dose Ricin | Timepoint:2h
SA1523263--5Low Dose Ricin | Timepoint:2h
SA1523273--6Low Dose Ricin | Timepoint:2h
SA1523283--9Low Dose Ricin | Timepoint:2h
SA1523293--8Low Dose Ricin | Timepoint:2h
SA1523303--7Low Dose Ricin | Timepoint:2h
SA1523313--10Low Dose Ricin | Timepoint:2h
SA1523328--3Low Dose Ricin | Timepoint:8h
SA1523338--7Low Dose Ricin | Timepoint:8h
SA1523348--8Low Dose Ricin | Timepoint:8h
SA1523358--9Low Dose Ricin | Timepoint:8h
SA1523368--6Low Dose Ricin | Timepoint:8h
SA1523378--5Low Dose Ricin | Timepoint:8h
SA1523388--1Low Dose Ricin | Timepoint:8h
SA1523398--2Low Dose Ricin | Timepoint:8h
SA1523408--4Low Dose Ricin | Timepoint:8h
SA1523414--1Low Dose Ricin | Timepoint:Overnight
SA1523424--2Low Dose Ricin | Timepoint:Overnight
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Collection:

Collection ID:CO001737
Collection Summary:Livers were removed from mice immediately following sacrifice and stored at -80 C. Before metabolite extraction, livers were homogenized into powder with mortar and pestle over liquid nitrogen.
Sample Type:Liver

Treatment:

Treatment ID:TR001757
Treatment Summary:Mice were injected via intraperitoneal (IP) route with either ricin or saline in 10 microliter per g body weight. Mice in all but the fasted group were given food and water ad libitum. Fasted mice were transferred to new cages with fresh bedding where they had food withheld while given water ad libitum. Mice were sacrificed at 2h (ricin), 8h (ricin and fasted), and overnight (16-22h, control, ricin, fasted).

Sample Preparation:

Sampleprep ID:SP001750
Sampleprep Summary:Microcentrifuge tubes with 30 mg of frozen liver tissue had 500 uL of ice-cold acetone and three Zirconia beads added to them. Tissues were lysed and homogenized, then sonicated and centrifuged. The supernatant was transferred to a new tube and 500 uL of ice-cold MeOH/H2O was added to the pellet. The pellet was sonicated and centrifuged again. The supernatant was combined with the acetone fraction and vacuum dried. Dried pellets were resuspended in acetonitrile/H2O with 1% Formic Acid, vortexed, sonicated, and centrifuged. 180 uL of eluate was used for LCMS analysis.

Combined analysis:

Analysis ID AN002719
Analysis type MS
Chromatography type HILIC
Chromatography system Agilent 1290
Column Waters Acquity BEH HILIC (150 x 2.1mm,1.7um)
MS Type ESI
MS instrument type QTOF
MS instrument name Agilent 6530 QTOF
Ion Mode POSITIVE
Units Intensity

Chromatography:

Chromatography ID:CH002007
Instrument Name:Agilent 1290
Column Name:Waters Acquity BEH HILIC (150 x 2.1mm,1.7um)
Column Pressure:600 bar
Column Temperature:40
Flow Gradient:100% B, 55% B, 20% B, 100%B
Flow Rate:0.400 mL/min
Retention Time:25 min
Solvent A:100% water; 0.1% formic acid
Solvent B:100% acetonitrile; 0.1% formic acid
Chromatography Type:HILIC

MS:

MS ID:MS002516
Analysis ID:AN002719
Instrument Name:Agilent 6530 QTOF
Instrument Type:QTOF
MS Type:ESI
MS Comments:mzML formatting acquired in ProteoWizard msconvert, Feature analysis performed in XCMS Online v3.7.1
Ion Mode:POSITIVE
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