Summary of Study ST002178

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 PR001386. The data can be accessed directly via it's Project DOI: 10.21228/M8K11W This work is supported by NIH grant, U2C- DK119886.

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Study IDST002178
Study TitleAge-independent Cardiac Protection by Pharmacological Activation of Beclin-1 During Endotoxemia and Its Association with Energy Metabolic Reprograming in Myocardium — A Targeted Metabolomics Study
Study SummaryBackground: We previously showed that Beclin-1-dependent autophagy is cardiac protective in a rodent model of endotoxemia using young adult mice. In this report, we compared the potential therapeutic effects of pharmacological Beclin-1 activating peptide, TB-peptide, on the cardiac outcomes of young adult and aged mice during endotoxemia. We further examined alterations in myocardial metabolism induced by lipopolysaccharide (LPS) challenge with and without the TB-peptide treatment. Methods and Results: C57BL/6J mice of 10-week and 24-month-old were challenged by LPS at doses at which cardiac dysfunction occurred. Following the treatment of TB-peptide or control vehicle, heart contractility, circulating cytokines, and myocardial autophagy were evaluated. A targeted metabolomics assay was applied to analyze cardiac metabolism. TB-peptide boosted autophagic response, attenuated cytokine production, and improved cardiac performance in both young and aged mice during endotoxemia. A targeted metabolomics assay was designed to detect a pool of 361 known metabolites, of which 156 were detected in at least one of the heart tissue samples. LPS-induced impairments were found in glucose and amino acid (AA) metabolisms in mice of all ages, and TB-peptide provided ameliorative effects to rescue these alterations. However, lipid metabolites were upregulated in the young group but moderately downregulated in the aged by LPS, suggesting an age-dependent response. TB-peptide mitigated LPS-mediated trend of lipids in the young mice but provided little effect on the aged ones. Conclusion: Pharmacological activation of Beclin-1 by TB-peptide protects the heart in both young and aged population during endotoxemia, suggest a therapeutic potential for sepsis-induced cardiomyopathy. Metabolomics analysis suggests that this age-independent protection by TB-peptide is associated with reprograming of energy production via glucose and AA metabolisms.
Institute
Loyola University Chicago Stritch School of Medicine
DepartmentSurgery
Last NameZang
First NameQun
Address2160 S. 1st Ave, Maywood, IL 60153
Emailqzang@luc.edu
Phone708-327-2472
Submit Date2022-05-23
Raw Data AvailableYes
Raw Data File Type(s)wiff
Analysis Type DetailLC-MS
Release Date2022-06-08
Release Version1
Qun Zang Qun Zang
https://dx.doi.org/10.21228/M8K11W
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:

Project:

Project ID:PR001386
Project DOI:doi: 10.21228/M8K11W
Project Title:Age-independent Cardiac Protection by Pharmacological Activation of Beclin-1 During Endotoxemia and Its Association with Energy Metabolic Reprograming in Myocardium — A Targeted Metabolomics Study
Project Type:A Targeted Metabolomics Study
Project Summary:Background: We previously showed that Beclin-1-dependent autophagy is cardiac protective in a rodent model of endotoxemia using young adult mice. In this report, we compared the potential therapeutic effects of pharmacological Beclin-1 activating peptide, TB-peptide, on the cardiac outcomes of young adult and aged mice during endotoxemia. We further examined alterations in myocardial metabolism induced by lipopolysaccharide (LPS) challenge with and without the TB-peptide treatment. Methods and Results: C57BL/6J mice of 10-week and 24-month-old were challenged by LPS at doses at which cardiac dysfunction occurred. Following the treatment of TB-peptide or control vehicle, heart contractility, circulating cytokines, and myocardial autophagy were evaluated. A targeted metabolomics assay was applied to analyze cardiac metabolism. TB-peptide boosted autophagic response, attenuated cytokine production, and improved cardiac performance in both young and aged mice during endotoxemia. A targeted metabolomics assay was designed to detect a pool of 361 known metabolites, of which 156 were detected in at least one of the heart tissue samples. LPS-induced impairments were found in glucose and amino acid (AA) metabolisms in mice of all ages, and TB-peptide provided ameliorative effects to rescue these alterations. However, lipid metabolites were upregulated in the young group but moderately downregulated in the aged by LPS, suggesting an age-dependent response. TB-peptide mitigated LPS-mediated trend of lipids in the young mice but provided little effect on the aged ones. Conclusion: Pharmacological activation of Beclin-1 by TB-peptide protects the heart in both young and aged population during endotoxemia, suggest a therapeutic potential for sepsis-induced cardiomyopathy. Metabolomics analysis suggests that this age-independent protection by TB-peptide is associated with reprograming of energy production via glucose and AA metabolisms.
Institute:Loyola University Chicago Stritch School of Medicine
Department:Surgery
Last Name:Zang
First Name:Qun
Address:2160 S. 1st Ave, Maywood, IL 60153
Email:qzang@luc.edu
Phone:708-327-2472
Funding Source:This work is supported by Nathan Shock Center Pilot Award (to Q.S.Z.), National Institute of Health grant 2R01GM111295-01 (to Q.S.Z.), HL109471 and CA215063 (to Z.P.L.), R01HL158515 and R01GM124108 (to J.L.), R01AG049494 (to D.P), American Heart Association grant AHA 19TP34910172 (to Z.P.L., AND NIH P30 AG013280 (to the University of Washington Nathan Shock Center). We also acknowledge The Northwest Metabolomics Research Center at the University of Washington, Seattle, and NIH S10 Grant 1S10OD021562-01 (to D.R) that funded a purchase of the LC-MS system used to acquire targeted metabolomics data.
Contributors:Matthew Kim, B.S, Azadeh Nikouee, PhD, Raymond Zou, Di Ren, PhD, Zhibin He, PhD, Ji Li, PhD, Lu Wang, PhD, Danijel Djukovic, PhD, Daniel Raftery, PhD, Hayley Purcell, B.S, Daniel Promislow, PhD, Yuxiao Sun, PhD, Mohammad Goodarzi, PhD, Qing-Jun Zhang, PhD, Zhi-Ping Liu, PhD, and Qun Sophia Zang, PhD

Subject:

Subject ID:SU002264
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Age Or Age Range:10-week and 24-month
Gender:Male

Factors:

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

mb_sample_id local_sample_id Age Group Treatment
SA2092082020-08-17_Zang-43_Box2Redo-Blank_NEGblank -
SA2092092020-08-17_Zang-43_Box2Redo-Blank_NEG (3)blank -
SA2092102020-08-17_Zang-43_Box2Redo-Blank_NEG (2)blank -
SA2092112020-08-17_Zang-43_Box2Redo-Blank_NEG (4)blank -
SA2092122020-08-17_Zang-43_Box2Redo-Blank_NEG (5)blank -
SA2092132020-08-17_Zang-43_Box2Redo-Blank_NEG (6)blank -
SA2092142020-08-17_Zang-43_Box2Redo-Blank0_NEGblank -
SA2092152020-08-17_Zang-43_Box2Redo-Blank_POSblank -
SA2092162020-08-17_Zang-43_Box2Redo-Blank_POS (6)blank -
SA2092172020-08-17_Zang-43_Box2Redo-Blank_POS (5)blank -
SA2092182020-08-17_Zang-43_Box2Redo-Blank_POS (4)blank -
SA2092192020-08-17_Zang-43_Box2Redo-Blank_POS (2)blank -
SA2092202020-08-17_Zang-43_Box2Redo-Blank_POS (3)blank -
SA2092212020-08-17_Zang-43_Box2Redo-Dummy_NEGinstrument quality control -
SA2092222020-08-17_Zang-43_Box2Redo-QC(I)#5_NEGinstrument quality control -
SA2092232020-08-17_Zang-43_Box2Redo-QC(I)#1_NEGinstrument quality control -
SA2092242020-08-17_Zang-43_Box2Redo-QC(I)#1_POSinstrument quality control -
SA2092252020-08-17_Zang-43_Box2Redo-Dummy_POSinstrument quality control -
SA2092262020-08-17_Zang-43_Box2Redo-QC(I)#3_POSinstrument quality control -
SA2092272020-08-17_Zang-43_Box2Redo-QC(I)#5_POSinstrument quality control -
SA2092282020-08-17_Zang-43_Box2Redo-QC(I)#4_POSinstrument quality control -
SA2092292020-08-17_Zang-43_Box2Redo-QC(I)#2_NEGinstrument quality control -
SA2092302020-08-17_Zang-43_Box2Redo-QC(I)#4_NEGinstrument quality control -
SA2092312020-08-17_Zang-43_Box2Redo-QC(I)#3_NEGinstrument quality control -
SA2092322020-08-17_Zang-43_Box2Redo-QC(I)#2_POSinstrument quality control -
SA2091222020-08-17_Zang-43_Box2Redo-28_POSOld Ctrl
SA2091232020-08-17_Zang-43_Box2Redo-21_POSOld Ctrl
SA2091242020-08-17_Zang-43_Box2Redo-34_POSOld Ctrl
SA2091252020-08-17_Zang-43_Box2Redo-23_NEGOld Ctrl
SA2091262020-08-17_Zang-43_Box2Redo-28_NEGOld Ctrl
SA2091272020-08-17_Zang-43_Box2Redo-34_NEGOld Ctrl
SA2091282020-08-17_Zang-43_Box2Redo-38_POSOld Ctrl
SA2091292020-08-17_Zang-43_Box2Redo-23_POSOld Ctrl
SA2091302020-08-17_Zang-43_Box2Redo-38_NEGOld Ctrl
SA2091312020-08-17_Zang-43_Box2Redo-21_NEGOld Ctrl
SA2091322020-08-17_Zang-43_Box2Redo-26_NEGOld LPS1
SA2091332020-08-17_Zang-43_Box2Redo-1_NEGOld LPS1
SA2091342020-08-17_Zang-43_Box2Redo-37_NEGOld LPS1
SA2091352020-08-17_Zang-43_Box2Redo-26_POSOld LPS1
SA2091362020-08-17_Zang-43_Box2Redo-40_NEGOld LPS1
SA2091372020-08-17_Zang-43_Box2Redo-7_NEGOld LPS1
SA2091382020-08-17_Zang-43_Box2Redo-37_POSOld LPS1
SA2091392020-08-17_Zang-43_Box2Redo-1_POSOld LPS1
SA2091402020-08-17_Zang-43_Box2Redo-40_POSOld LPS1
SA2091412020-08-17_Zang-43_Box2Redo-7_POSOld LPS1
SA2091502020-08-17_Zang-43_Box2Redo-16_NEGOld LPS1_peptide
SA2091512020-08-17_Zang-43_Box2Redo-16_POSOld LPS1_peptide
SA2091422020-08-17_Zang-43_Box2Redo-27_NEGOld LPS1-peptide
SA2091432020-08-17_Zang-43_Box2Redo-27_POSOld LPS1-peptide
SA2091442020-08-17_Zang-43_Box2Redo-32_POSOld LPS1-peptide
SA2091452020-08-17_Zang-43_Box2Redo-39_POSOld LPS1-peptide
SA2091462020-08-17_Zang-43_Box2Redo-39_NEGOld LPS1-peptide
SA2091472020-08-17_Zang-43_Box2Redo-10_NEGOld LPS1-peptide
SA2091482020-08-17_Zang-43_Box2Redo-32_NEGOld LPS1-peptide
SA2091492020-08-17_Zang-43_Box2Redo-10_POSOld LPS1-peptide
SA2091522020-08-17_Zang-43_Box2Redo-43_NEGOld peptide
SA2091532020-08-17_Zang-43_Box2Redo-8_NEGOld peptide
SA2091542020-08-17_Zang-43_Box2Redo-3_NEGOld peptide
SA2091552020-08-17_Zang-43_Box2Redo-3_POSOld peptide
SA2091562020-08-17_Zang-43_Box2Redo-29_POSOld peptide
SA2091572020-08-17_Zang-43_Box2Redo-43_POSOld peptide
SA2091582020-08-17_Zang-43_Box2Redo-8_POSOld peptide
SA2091592020-08-17_Zang-43_Box2Redo-29_NEGOld peptide
SA2092332020-08-17_Zang-43_Box2Redo-QC(S)#3_POSsample quality control -
SA2092342020-08-17_Zang-43_Box2Redo-QC(S)#1_POSsample quality control -
SA2092352020-08-17_Zang-43_Box2Redo-QC(S)#2_POSsample quality control -
SA2092362020-08-17_Zang-43_Box2Redo-QC(S)#5_NEGsample quality control -
SA2092372020-08-17_Zang-43_Box2Redo-QC(S)#2_NEGsample quality control -
SA2092382020-08-17_Zang-43_Box2Redo-QC(S)#3_NEGsample quality control -
SA2092392020-08-17_Zang-43_Box2Redo-QC(S)#1_NEGsample quality control -
SA2092402020-08-17_Zang-43_Box2Redo-QC(S)#4_NEGsample quality control -
SA2092412020-08-17_Zang-43_Box2Redo-QC(S)#5_POSsample quality control -
SA2092422020-08-17_Zang-43_Box2Redo-QC(S)#4_POSsample quality control -
SA2091212020-08-17_Zang-43_Box2Redo-Blank0_POS- -
SA2091602020-08-17_Zang-43_Box2Redo-17_NEGYoung Ctrl
SA2091612020-08-17_Zang-43_Box2Redo-30_NEGYoung Ctrl
SA2091622020-08-17_Zang-43_Box2Redo-19_NEGYoung Ctrl
SA2091632020-08-17_Zang-43_Box2Redo-5_POSYoung Ctrl
SA2091642020-08-17_Zang-43_Box2Redo-9_POSYoung Ctrl
SA2091652020-08-17_Zang-43_Box2Redo-19_POSYoung Ctrl
SA2091662020-08-17_Zang-43_Box2Redo-9_NEGYoung Ctrl
SA2091672020-08-17_Zang-43_Box2Redo-31_NEGYoung Ctrl
SA2091682020-08-17_Zang-43_Box2Redo-5_NEGYoung Ctrl
SA2091692020-08-17_Zang-43_Box2Redo-30_POSYoung Ctrl
SA2091702020-08-17_Zang-43_Box2Redo-31_POSYoung Ctrl
SA2091712020-08-17_Zang-43_Box2Redo-17_POSYoung Ctrl
SA2091722020-08-17_Zang-43_Box2Redo-14_NEGYoung LPS3
SA2091732020-08-17_Zang-43_Box2Redo-6_NEGYoung LPS3
SA2091742020-08-17_Zang-43_Box2Redo-4_NEGYoung LPS3
SA2091752020-08-17_Zang-43_Box2Redo-11_NEGYoung LPS3
SA2091762020-08-17_Zang-43_Box2Redo-25_NEGYoung LPS3
SA2091772020-08-17_Zang-43_Box2Redo-36_POSYoung LPS3
SA2091782020-08-17_Zang-43_Box2Redo-36_NEGYoung LPS3
SA2091792020-08-17_Zang-43_Box2Redo-11_POSYoung LPS3
SA2091802020-08-17_Zang-43_Box2Redo-14_POSYoung LPS3
SA2091812020-08-17_Zang-43_Box2Redo-6_POSYoung LPS3
SA2091822020-08-17_Zang-43_Box2Redo-25_POSYoung LPS3
SA2091832020-08-17_Zang-43_Box2Redo-4_POSYoung LPS3
SA2091842020-08-17_Zang-43_Box2Redo-35_NEGYoung LPS3_peptide
SA2091852020-08-17_Zang-43_Box2Redo-20_POSYoung LPS3_peptide
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Collection:

Collection ID:CO002257
Collection Summary:Wild-type (WT) C57BL/6 mice were used in this study. Endotoxemia was induced in young (10-week) and aged (24-week) male mice by lipopolysaccharide (LPS). Based on published results as well as observations in our laboratory, male and female mice showed significantly different susceptibility to systemic symptoms in sepsis models. Thus, male but not female mice were chosen for the experiments presented in this report. LPS was administered intraperitoneally (i.p.), and mice were weighed individually to determine the exact amount of LPS (MilliporeSigma, Burlington, MA; catalog number L3012) required to achieve the doses indicated in the figure legends. Sterile endotoxin-free PBS was used as a vehicle control in sham groups. In some experiments, Beclin-1-activating peptide (TB peptide), synthesized according to a published sequence24 by NonoPep (Shanghai, China), was administered i.p. at a dose of 16 mg/kg in 100μl of PBS 30 minutes post LPS-challenge. Heart tissue was harvested 18 hours post LPS challenge.
Collection Protocol Filename:Zang_Protocol.docx
Sample Type:Heart
Storage Conditions:-80℃

Treatment:

Treatment ID:TR002276
Treatment Summary:Endotoxemia was induced in young (10-week) and aged (24-week) male mice by lipopolysaccharide (LPS). Based on published results as well as observations in our laboratory, male and female mice showed significantly different susceptibility to systemic symptoms in sepsis models. Thus, male but not female mice were chosen for the experiments presented in this report. LPS was administered intraperitoneally (i.p.), and mice were weighed individually to determine the exact amount of LPS (MilliporeSigma, Burlington, MA; catalog number L3012) required to achieve the required doses. Sterile endotoxin-free PBS was used as a vehicle control in sham groups. Consistent with literature and as expected, we observed that older mice were more susceptible to the toxic effects induced by LPS. 24-month-old (aged) mice showed impaired cardiac function but were able to survive when receiving LPS challenged at 1mg/kg. However, greater fatality was observed when LPS dose was increased to 3 mg/kg. In 10-week-old (young adult) mice, 3 mg/kg LPS triggered heart dysfunction without impact on survival, whereases at 10 mg/kg, we observed significant LPS-induced fatality in the group. Due to the different sensitivities to LPS between the aged and young adult mice, we were not able to choose a universal dose of LPS to induce cardiac dysfunction and to perform follow up analysis in both groups. Therefore, we used the physiological function of the heart as a base for comparison in the studies performed in this report. Our previous research provided evidence that stimulating beclin-1 dependent autophagy improves cardiac performance during endotoxemia in young adult mice, and thus Beclin-1-activating peptide (TB peptide) holds a promising therapeutic potential for sepsis. In this report, we examined whether TB-peptide exerts a similar protective effect on aged animals under the same condition. In our experimental setting, sham or LPS challenge was administered to groups of 24-month-old and 10-week-old mice followed by treatment with TB-peptide, administered i.p. at a dose of 16 mg/kg in 100μl of PBS 30 minutes post LPS-challenge. Heart tissue was collected 18 hours post LPS challenge.

Sample Preparation:

Sampleprep ID:SP002270
Sampleprep Summary:Sample preparation: aqueous metabolites were extracted using a methanol-based protein precipitation method as described previously. Briefly, heart tissue samples were homogenized in cold water using zirconium oxide beads, methanol was added, and samples were vortexed and then stored for 30 minutes at -20˚C. Afterwards, samples were first sonicated in an ice bath for 10 minutes, centrifuged for 15 min at 18,000g and 4˚C, and then a fixed volume of supernatant was collected from each sample. Lastly, recovered supernatants were dried on a SpeedVac and reconstituted for LC-MS analysis. Protein pallets that were left over from the sample prep were saved for BCA assay. (2) LC-MS analysis: samples were analyzed on a duplex-LC-MS system composed of two Shimadzu UPLC pumps, CTC Analytics PAL HTC-xt temperature-controlled auto-sampler and AB Sciex 6500+ Triple Quadrupole MS equipped with ESI ionization source. UPLC pumps were connected to the auto-sampler in parallel and were able to perform two chromatography separations independently from each other. Each sample was injected twice on two identical analytical columns (Waters XBridge BEH Amide XP) performing separations in hydrophilic interaction liquid chromatography (HILIC) mode. While one column was performing separation and MS data acquisition in ESI(+) ionization mode, the other column was getting equilibrated for sample injection, chromatography separation and MS data acquisition in ESI(-) mode. Each chromatography separation was 18 minutes (total analysis time per sample was 36 minutes).

Combined analysis:

Analysis ID AN003566 AN003567
Analysis type MS MS
Chromatography type HILIC HILIC
Chromatography system Shimadzu Nexera X2 Shimadzu Nexera X2
Column Waters XBridge BEH Amide (150 x 2.1mm,2.5um) Waters XBridge BEH Amide (150 x 2.1mm,2.5um)
MS Type ESI ESI
MS instrument type QTRAP QTRAP
MS instrument name ABI Sciex 6500+ ABI Sciex 6500+
Ion Mode POSITIVE NEGATIVE
Units count per second (cps) count per second (CPS)

Chromatography:

Chromatography ID:CH002637
Instrument Name:Shimadzu Nexera X2
Column Name:Waters XBridge BEH Amide (150 x 2.1mm,2.5um)
Chromatography Type:HILIC
  
Chromatography ID:CH002638
Instrument Name:Shimadzu Nexera X2
Column Name:Waters XBridge BEH Amide (150 x 2.1mm,2.5um)
Chromatography Type:HILIC

MS:

MS ID:MS003323
Analysis ID:AN003566
Instrument Name:ABI Sciex 6500+
Instrument Type:QTRAP
MS Type:ESI
MS Comments:MS data acquisition was performed in multiple-reaction-monitoring (MRM) mode. The whole LC-MS system was controlled using AB Sciex Analyst 1.6.3 software. Measured MS peaks were integrated using AB Sciex MultiQuant 3.0.3 software. In addition to the study samples, two sets of quality control (QC) samples were used to monitor the assay performance as well as data reproducibility. One QC was a pooled human serum sample used to monitor system performance and the other QC was pooled study samples and this QC was used to monitor data reproducibility. Isotope labeled compounds were used to monitor sample preparation and injection. Highly reproducible MS data were generated, having an average coefficient of variances (CVs) among the metabolites of 5.6%. Data for each sample were normalized according to bicinchoninic acid (BCA)-based quantification of total protein count.
Ion Mode:POSITIVE
Analysis Protocol File:Zang_Protocol.docx
  
MS ID:MS003324
Analysis ID:AN003567
Instrument Name:ABI Sciex 6500+
Instrument Type:QTRAP
MS Type:ESI
MS Comments:MS data acquisition was performed in multiple-reaction-monitoring (MRM) mode. The whole LC-MS system was controlled using AB Sciex Analyst 1.6.3 software. Measured MS peaks were integrated using AB Sciex MultiQuant 3.0.3 software. In addition to the study samples, two sets of quality control (QC) samples were used to monitor the assay performance as well as data reproducibility. One QC was a pooled human serum sample used to monitor system performance and the other QC was pooled study samples and this QC was used to monitor data reproducibility. Isotope labeled compounds were used to monitor sample preparation and injection. Highly reproducible MS data were generated, having an average coefficient of variances (CVs) among the metabolites of 5.6%. Data for each sample were normalized according to bicinchoninic acid (BCA)-based quantification of total protein count.
Ion Mode:NEGATIVE
Analysis Protocol File:Zang_Protocol.docx
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