Summary of Study ST002228

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 PR001419. The data can be accessed directly via it's Project DOI: 10.21228/M89D8V 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	ST002228
Study Title	Estrogen receptor a deficiency in cardiac myocytes reprograms heart-derived extracellular vesicle proteome and induces obesity in female mice (Part 2)
Study Summary	Dysregulation of ERα has been linked with increased metabolic and cardiovascular disease risk. Uncovering the impact of ERα deficiency in specific tissues has implications for understanding the role of ERα in normal physiology and disease, the increased disease risk in postmenopausal women, and the design of tissue-specific ERα-based therapies for a range of pathologies including cardiac disease and cancer. Cardiac myocyte-specific ER knockout mice (ERαHKO) were generated to assess the role of ERα in the heart. Female ERαHKO mice displayed a modest cardiac phenotype, but unexpectedly, the most striking phenotype was obesity in female ERαHKO but not male ERHKO mice. In female ERαHKO mice we identified cardiac dysfunction, mild glucose and insulin intolerance, and reduced ERα gene expression in skeletal muscle and white adipose tissue (WAT). Gene expression, protein, lipidomic and metabolomic analyses showed evidence of contractile and/or metabolic dysregulation in heart, skeletal muscle and WAT. We also show that extracellular vesicles (EVs) collected from the perfusate of isolated hearts from female ERαHKO mice have a distinct proteome, and these EVs have the capacity to reprogram the proteome of a skeletal muscle cell including proteins linked with ERα, fatty acid regulation, lipid metabolism and mitochondrial function. This study uncovers a cardiac-initiated and sex-specific cardiometabolic phenotype that is regulated by ERα.
Institute	Baker Heart and Diabetes Institute
Department	Discovery and Preclinical Science
Laboratory	Cardiac Hypertrophy
Last Name	Tham
First Name	Yow Keat
Address	75 Commercial Rd, Melbourne, Victoria, 3004, Australia
Email	yowkeat.tham@baker.edu.au
Phone	0385321266
Submit Date	2022-07-17
Raw Data Available	Yes
Raw Data File Type(s)	qgd
Analysis Type Detail	GC-MS
Release Date	2023-01-02
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR001419
Project DOI:	doi: 10.21228/M89D8V
Project Title:	Estrogen receptor α deficiency in cardiac myocytes reprograms heart-derived extracellular vesicle proteome and induces obesity in female mice
Project Summary:	Tissues (ventricles, skeletal muscles-soleus, subcutaneous fat) from male and female ERalpha cardiac-specific knockout and floxed control aged mice (54-59 weeks old, male FC n=5, male KO n=5, female FC n=8, female KO n=7 ) were subjected to metabolomic profiling.
Institute:	Baker Heart and Diabetes Institute
Department:	Discovery and Preclinical Science
Laboratory:	Cardiac Hypertrophy
Last Name:	Tham
First Name:	Yow Keat
Address:	75 Commercial Rd, Melbourne, Victoria, 3004, Australia
Email:	yowkeat.tham@baker.edu.au
Phone:	0385321266
Publications:	https://www.nature.com/articles/s44161-023-00223-z

Subject:

Subject ID:	SU002314
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	C57BL6 and FVB mixed strain
Age Or Age Range:	54-59 weeks old
Gender:	Male and female
Animal Feed:	Specialty Feeds Irradiated Rat and Mouse Standard Chow Diet

Factors:

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

mb_sample_id	local_sample_id	Sex	Genotype	Tissue
SA212361	37910_Sk	Female	ERalpha-knockout	Skeletal muscle
SA212362	37905_Sk	Female	ERalpha-knockout	Skeletal muscle
SA212363	37899_Sk	Female	ERalpha-knockout	Skeletal muscle
SA212364	37906_Sk	Female	ERalpha-knockout	Skeletal muscle
SA212365	37917_Sk	Female	ERalpha-knockout	Skeletal muscle
SA212367	37914_Sk	Female	ERalpha-knockout	Skeletal muscle
SA212368	37915_Sk	Female	ERalpha-knockout	Skeletal muscle
SA212369	37910_sub	Female	ERalpha-knockout	Subcutaneous fat
SA212370	37906_sub	Female	ERalpha-knockout	Subcutaneous fat
SA212371	37914_sub	Female	ERalpha-knockout	Subcutaneous fat
SA212372	37915_sub	Female	ERalpha-knockout	Subcutaneous fat
SA212373	37917_sub	Female	ERalpha-knockout	Subcutaneous fat
SA212374	37899_sub	Female	ERalpha-knockout	Subcutaneous fat
SA212375	37905_sub	Female	ERalpha-knockout	Subcutaneous fat
SA212355	37910_H	Female	ERalpha-knockout	Ventricles
SA212356	37914_H	Female	ERalpha-knockout	Ventricles
SA212357	37917_H	Female	ERalpha-knockout	Ventricles
SA212358	37906_H	Female	ERalpha-knockout	Ventricles
SA212359	37905_H	Female	ERalpha-knockout	Ventricles
SA212360	37915_H	Female	ERalpha-knockout	Ventricles
SA212366	37899_H	Female	ERalpha-knockout	Ventricles
SA212376	37913_Sk	Female	Floxed Control	Skeletal muscle
SA212378	37912_Sk	Female	Floxed Control	Skeletal muscle
SA212379	37916_Sk	Female	Floxed Control	Skeletal muscle
SA212380	37896_Sk	Female	Floxed Control	Skeletal muscle
SA212381	37909_Sk	Female	Floxed Control	Skeletal muscle
SA212382	37891_Sk	Female	Floxed Control	Skeletal muscle
SA212390	37893_Sk	Female	Floxed Control	Skeletal muscle
SA212399	37890_Sk	Female	Floxed Control	Skeletal muscle
SA212377	37890_sub	Female	Floxed Control	Subcutaneous fat
SA212383	37893_sub	Female	Floxed Control	Subcutaneous fat
SA212384	37916_sub	Female	Floxed Control	Subcutaneous fat
SA212385	37913_sub	Female	Floxed Control	Subcutaneous fat
SA212386	37891_sub	Female	Floxed Control	Subcutaneous fat
SA212387	37909_sub	Female	Floxed Control	Subcutaneous fat
SA212388	37896_sub	Female	Floxed Control	Subcutaneous fat
SA212389	37912_sub	Female	Floxed Control	Subcutaneous fat
SA212391	37896_H	Female	Floxed Control	Ventricles
SA212392	37893_H	Female	Floxed Control	Ventricles
SA212393	37909_H	Female	Floxed Control	Ventricles
SA212394	37913_H	Female	Floxed Control	Ventricles
SA212395	37912_H	Female	Floxed Control	Ventricles
SA212396	37916_H	Female	Floxed Control	Ventricles
SA212397	37890_H	Female	Floxed Control	Ventricles
SA212398	37891_H	Female	Floxed Control	Ventricles
SA212410	37904_Sk	Male	ERalpha-knockout	Skeletal muscle
SA212411	37902_Sk	Male	ERalpha-knockout	Skeletal muscle
SA212412	37901_Sk	Male	ERalpha-knockout	Skeletal muscle
SA212413	37898_Sk	Male	ERalpha-knockout	Skeletal muscle
SA212414	37908_Sk	Male	ERalpha-knockout	Skeletal muscle
SA212400	37908_sub	Male	ERalpha-knockout	Subcutaneous fat
SA212401	37904_sub	Male	ERalpha-knockout	Subcutaneous fat
SA212402	37902_sub	Male	ERalpha-knockout	Subcutaneous fat
SA212408	37898_sub	Male	ERalpha-knockout	Subcutaneous fat
SA212409	37901_sub	Male	ERalpha-knockout	Subcutaneous fat
SA212403	37898_H	Male	ERalpha-knockout	Ventricles
SA212404	37901_H	Male	ERalpha-knockout	Ventricles
SA212405	37908_H	Male	ERalpha-knockout	Ventricles
SA212406	37904_H	Male	ERalpha-knockout	Ventricles
SA212407	37902_H	Male	ERalpha-knockout	Ventricles
SA212419	37907_Sk	Male	Floxed Control	Skeletal muscle
SA212420	37900_Sk	Male	Floxed Control	Skeletal muscle
SA212426	37897_Sk	Male	Floxed Control	Skeletal muscle
SA212427	37895_Sk	Male	Floxed Control	Skeletal muscle
SA212428	37894_Sk	Male	Floxed Control	Skeletal muscle
SA212421	37897_sub	Male	Floxed Control	Subcutaneous fat
SA212422	37900_sub	Male	Floxed Control	Subcutaneous fat
SA212423	37907_sub	Male	Floxed Control	Subcutaneous fat
SA212424	37895_sub	Male	Floxed Control	Subcutaneous fat
SA212425	37894_sub	Male	Floxed Control	Subcutaneous fat
SA212415	37907_H	Male	Floxed Control	Ventricles
SA212416	37895_H	Male	Floxed Control	Ventricles
SA212417	37897_H	Male	Floxed Control	Ventricles
SA212418	37900_H	Male	Floxed Control	Ventricles
SA212429	37894_H	Male	Floxed Control	Ventricles

Showing results 1 to 75 of 75

Showing results 1 to 75 of 75

Collection:

Collection ID:	CO002307
Collection Summary:	Ventricles were dissected from mice such that each sample will include left and right ventricular tissue.Tissues were snap frozen in liquid nitrogen and stored in -80 freezer until tissues were processed for metabolomics.
Sample Type:	Ventricles, Skeletal Muscles, Subcutaneous fat
Storage Conditions:	-80℃

Treatment:

Treatment ID:	TR002326
Treatment Summary:	Mice did not undergo specific treatment, as this was a basal phenotyping study. Mice were fasted for 6 hours before dissections, and a lethal dose of anesthesia was delivered via intraperitoneal injection before tissue collection.
Animal Anesthesia:	Pentobarbitone
Animal Fasting:	6 hours

Sample Preparation:

Sampleprep ID:	SP002320
Sampleprep Summary:	600 µL of 3:1 methanol/water (containing 13C6 Sorbitol & 13C5, 15N Valine as internal standards) was added to the samples and cryomilled for 45 sec x 3 at 6800 rpm. 480 µL of the homogenate was transferred into a fresh tube and centrifuged for 10 min at 12700 rpm. 100 µL of the supernatant of each sample was pooled to make up the pooled biological quality controls (PBQCs). 100 µL of each sample (study samples & PBQCs) was dried using a SpeedVac vacuum concentrator prior to derivatization. The samples and PBQCs were methoximated (methoxyamine hydrochloride - 25 µL) and trimethylsilylated (BSTFA - 25 µL) to enable gas phase analysis.

Sampleprep ID:

SP002320

Sampleprep Summary:

600 µL of 3:1 methanol/water (containing 13C6 Sorbitol & 13C5, 15N Valine as internal standards) was added to the samples and cryomilled for 45 sec x 3 at 6800 rpm. 480 µL of the homogenate was transferred into a fresh tube and centrifuged for 10 min at 12700 rpm. 100 µL of the supernatant of each sample was pooled to make up the pooled biological quality controls (PBQCs). 100 µL of each sample (study samples & PBQCs) was dried using a SpeedVac vacuum concentrator prior to derivatization. The samples and PBQCs were methoximated (methoxyamine hydrochloride - 25 µL) and trimethylsilylated (BSTFA - 25 µL) to enable gas phase analysis.

Combined analysis:

Analysis ID	AN003637
Analysis type	MS
Chromatography type	GC
Chromatography system	Shimadzu Nexis GC-2030
Column	Agilent DB5-MS (30m x 0.25mm, 0.25um)
MS Type	EI
MS instrument type	Triple quadrupole
MS instrument name	Shimadzu TQ8050 NX
Ion Mode	POSITIVE
Units	abundance

Chromatography:

Chromatography ID:	CH002692
Chromatography Summary:	The GC-MS system used comprised of an AOC6000 autosampler, a 2030 Shimadzu gas chromatograph and a TQ8050NX triple quadrupole mass spectrometer (Shimadzu, Japan). The mass spectrometer was tuned according to the manufacturer’s recommendations using tris-(perfluorobutyl)-amine (CF43). GC-MS was performed on a 30m Agilent DB-5 column with 0.25mm internal diameter column and 1µm film thickness. The injection temperature (inlet) was set at 280°C, the MS transfer line at 280°C and the ion source adjusted to 200°C. Helium was used as the carrier gas at a flow rate of 1 mL/min and argon gas was used in the collision cell to generate the MRM product ion. The analysis of TMS samples was performed under the following oven temperature program; 100°C start temperature, hold for 4 minutes, followed by a 10°C min-1 oven temperature ramp to 320°C with a following final hold for 11 minutes. Approximately 520 targets were collected using the Shimadzu Smart Metabolite Database, where each target comprised a quantifier MRM along with a qualifier MRM, which covers approximately 350 endogenous metabolites and multiple stable isotopically labelled internal standards. Resultant data was processed using Shimadzu LabSolutions Insight software, where peak integrations were visually validated and manually corrected where required.
Instrument Name:	Shimadzu Nexis GC-2030
Column Name:	Agilent DB5-MS (30m x 0.25mm, 0.25um)
Chromatography Type:	GC

MS:

MS ID:	MS003388
Analysis ID:	AN003637
Instrument Name:	Shimadzu TQ8050 NX
Instrument Type:	Triple quadrupole
MS Type:	EI
MS Comments:	The GC-MS system used comprised of an AOC6000 autosampler, a 2030 Shimadzu gas chromatograph and a TQ8050NX triple quadrupole mass spectrometer (Shimadzu, Japan). The mass spectrometer was tuned according to the manufacturer’s recommendations using tris-(perfluorobutyl)-amine (CF43). GC-MS was performed on a 30m Agilent DB-5 column with 0.25mm internal diameter column and 1µm film thickness. The injection temperature (inlet) was set at 280°C, the MS transfer line at 280°C and the ion source adjusted to 200°C. Helium was used as the carrier gas at a flow rate of 1 mL/min and argon gas was used in the collision cell to generate the MRM product ion. The analysis of TMS samples was performed under the following oven temperature program; 100°C start temperature, hold for 4 minutes, followed by a 10°C min-1 oven temperature ramp to 320°C with a following final hold for 11 minutes. Approximately 520 targets were collected using the Shimadzu Smart Metabolite Database, where each target comprised a quantifier MRM along with a qualifier MRM, which covers approximately 350 endogenous metabolites and multiple stable isotopically labelled internal standards. Resultant data was processed using Shimadzu LabSolutions Insight software, where peak integrations were visually validated and manually corrected where required.
Ion Mode:	POSITIVE