Summary of Study ST002204
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 PR001407. The data can be accessed directly via it's Project DOI: 10.21228/M8V995 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 | ST002204 |
| Study Title | Endothelial Sirtuin1 Suppresses Whole-body Insulin Sensitivity by Modulating the Secretome |
| Study Type | End point |
| Study Summary | Sirtuin1 (Sirt1) in skeletal muscle (SK) and fat protects against metabolic damage by stimulating insulin sensitivity. Here we report that mice with selective deletion of endothelial Sirt1 (E-Sirt1-KO) paradoxically exhibit heightened whole-body insulin sensitivity. Akt phosphorylation, glucose uptake, and glycolysis are boosted in SK and brown adipose tissue (BAT) of E-Sirt1-KO mice. E-Sirt1-KO mice have higher energy expenditure and are partially protected from high-fat diet-induced insulin resistance. Enhanced insulin sensitivity and peripheral tissue Akt phosphorylation in E-Sirt1-KO mice is transferrable to wild-type mice via the systemic circulation after surgical parabiosis. Silencing of Sirt1 in endothelial cells upregulates transcription of the F-actin-binding protein thymosin beta-4 (Tβ4), whose secretion activates Akt in skeletal myotubes. Sirt1 downregulation stimulates endothelial Tβ4 transcription through inhibition of autophagy and upregulation of nuclear factor-kappa B signaling. Thus, unlike Sirt1 in skeletal muscle and fat, endothelial Sirt1 curtails whole-body insulin sensitivity by inhibiting expression of secreted Tβ4 |
| Institute | University of Iowa |
| Department | Internal medicine |
| Laboratory | irani |
| Last Name | Irani |
| First Name | Kaikobad |
| Address | RM 2256 CBRB, Newton Rd, Iowa City, IA 52242 |
| kaikobad-irani@uiowa.edu | |
| Phone | 3193358821 |
| Submit Date | 2021-12-02 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | cdf |
| Analysis Type Detail | GC-MS |
| Release Date | 2022-07-22 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001407 |
| Project DOI: | doi: 10.21228/M8V995 |
| Project Title: | Endothelial Sirtuin1 Suppresses Whole-body Insulin Sensitivity by Modulating the Secretome |
| Project Type: | Profiling metabolites in skeletal muscle |
| Project Summary: | Profiling metabolites in skeletal muscle from wild type vs. endothelium-specific Sirt1 knockout (E-Sirt1-KO) mice. |
| Institute: | University of Iowa |
| Department: | Internal medicine |
| Laboratory: | Irani lab |
| Last Name: | Irani |
| First Name: | Kaikobad |
| Address: | RM 2256 CBRB, Newton Rd, Iowa City, IA 52242 |
| Email: | kaikobad-irani@uiowa.edu |
| Phone: | 319-335-8821 |
| Funding Source: | AHA Award #20CDA35320042 |
Subject:
| Subject ID: | SU002290 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | wild type and endothelium-specific Sirt1 knockout mice |
| Age Or Age Range: | 12 weeks old |
| Weight Or Weight Range: | ~30 g |
| Height Or Height Range: | n/a |
| Gender: | Male |
| Animal Animal Supplier: | Jax |
| Animal Housing: | University of Iowa animal house |
| Animal Light Cycle: | 12 h light/12 h night |
| Animal Feed: | Standard Chow |
| Animal Water: | Tap water |
| Animal Inclusion Criteria: | n/a |
| Species Group: | n/a |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Genotype |
|---|---|---|
| SA211362 | S7 | KO |
| SA211363 | S8 | KO |
| SA211364 | S9 | KO |
| SA211365 | S6 | KO |
| SA211366 | S5 | KO |
| SA211367 | S4 | KO |
| SA211368 | S2 | WT |
| SA211369 | S3 | WT |
| SA211370 | S1 | WT |
| Showing results 1 to 9 of 9 |
Collection:
| Collection ID: | CO002283 |
| Collection Summary: | 40 mg of tibialis anterior was rapidly harvested and frozen immediately in liquid nitrogen. Metabolites were separated, measured, and analyzed. |
| Sample Type: | Muscle |
Treatment:
| Treatment ID: | TR002302 |
| Treatment Summary: | After 5 to 6 h of fasting, mice were anesthetized and skeletal muscle were collected. |
Sample Preparation:
| Sampleprep ID: | SP002296 |
| Sampleprep Summary: | Tissue samples were lyophilized overnight prior to bead mill homogenization in 18:1 (µl:mg wet tissue weight) ice-cold 2:2:1 methanol:acetonitrile:water extraction buffer containing a mixture of internal standards (D4-citric acid, D4-succinic acid, D8-valine, and U13C-labeled glutamine, glutamic acid, lysine, methionine, serine, and tryptophan; Cambridge Isotope Laboratories). Homogenates were rotated for 1 hour at -20°C. Homogenates were centrifuged for 10 minutes at 21,000 x g, and 150 µl of the cleared metabolite extracts were transferred to autosampler vials and dried using a SpeedVac vacuum concentrator (Thermo). Dried metabolite extracts were reconstituted in 30 μl of 11.4 mg/ml methoxyamine (MOX) in anhydrous pyridine, vortexed for 5 minutes, and heated for 1 hour at 60°C. Next, to each sample 20 μl of N,O-Bis(trimethylsilyl)trifluoroacetamide (TMS) was added, samples were vortexed for 1 minute, and heated for 30 minutes at 60°C. |
| Sampleprep Protocol Filename: | GC_Method_Tissue.pdf |
Combined analysis:
| Analysis ID | AN003607 |
|---|---|
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | Thermo Trace 1310 |
| Column | Thermo TraceGold TG-5SilMS |
| MS Type | EI |
| MS instrument type | Single quadrupole |
| MS instrument name | Thermo ISQ |
| Ion Mode | POSITIVE |
| Units | AU |
Chromatography:
| Chromatography ID: | CH002666 |
| Chromatography Summary: | 1 μl of derivatized sample was injected into a Trace 1300 GC (Thermo) fitted with a TraceGold TG-5SilMS column (Thermo) operating under the following conditions: split ratio = 20-1, split flow = 24 μl/minute, purge flow = 5 ml/minute, carrier mode = Constant Flow, and carrier flow rate = 1.2 ml/minute. The GC oven temperature gradient was as follows: 80°C for 3 minutes, increasing at a rate of 20°C/minute to 280°C, and holding at a temperature at 280°C for 8 minutes. |
| Methods Filename: | GC_Method_Tissue.pdf |
| Instrument Name: | Thermo Trace 1310 |
| Column Name: | Thermo TraceGold TG-5SilMS |
| Chromatography Type: | GC |
MS:
| MS ID: | MS003361 |
| Analysis ID: | AN003607 |
| Instrument Name: | Thermo ISQ |
| Instrument Type: | Single quadrupole |
| MS Type: | EI |
| MS Comments: | The MS was operated from 3.90 to 21.00 minutes in EI mode (-70eV) using select ion monitoring (SIM). Data were QC normalized using NOREVA (PMID: 28525573). |
| Ion Mode: | POSITIVE |
