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.

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.

Study ID	ST002178
Study 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
Study 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
Submit Date	2022-05-23
Raw Data Available	Yes
Raw Data File Type(s)	wiff
Analysis Type Detail	LC-MS
Release Date	2022-06-08
Release Version	1

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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).

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).