Summary of Study ST003927

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 PR002460. The data can be accessed directly via it's Project DOI: 10.21228/M8NR8D 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	ST003927
Study Title	Histidine decarboxylase inhibition attenuates cancer-associated muscle wasting
Study Summary	Cancer cachexia is a multifactorial syndrome involving muscle and fat wasting, inflammation, and metabolic dysfunction. Across cancer subtypes, pancreatic cancer has one of the highest cachexia incidence rates at approximately 80%. Given the advanced age of most pancreatic cancer patients, we sought to query cancer-associated muscle wasting using an age-matched murine model. We found that histamine and histamine decarboxylase (HDC) activity were specifically elevated in the muscles of aged tumor-bearing mice. We further found that 1) wasting stimuli induced histamine production and enhanced HDC activity; 2) exogenous histamine was sufficient to induce atrophy-associated gene expression; 3) inhibition of HDC activity by α-fluoromethylhistidine (FMH) protected against atrophy; 4) treatment of tumor-bearing mice with FMH rescued muscle wasting; and 5) a calcineurin inhibitor was able to rescue histamine-associated increases in calcium/atrogene signaling. In summary, we present a novel metabolic pathway that has significant implications for the treatment of cachectic cancer patients
Institute	Indiana University School of Medicine
Last Name	Doles
First Name	Jason
Address	635 Barnhill Drive
Email	jadoles@iu.edu
Phone	4027080332
Submit Date	2025-05-22
Analysis Type Detail	LC-MS
Release Date	2025-06-17
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR002460
Project DOI:	doi: 10.21228/M8NR8D
Project Title:	Muscle metabolome of young/age healthy/tumor bearing mice
Project Summary:	The goal of the study is to evaluate the metabolome differences in the muscles of young and aged mice when implanted with KPC cells (pancreatic cancer cells). We focused on the differential metabolite abundance between the young and the aged tumor bearing mice. We found histamine was significantly upregulated in the aged tumor-bearing mice. We followed this data and found a histamine signaling axis in cachexia.
Institute:	Indiana University School of Medicine
Last Name:	Doles
First Name:	Jason
Address:	635 Barnhill Drive
Email:	jadoles@iu.edu
Phone:	4027080332

Subject:

Subject ID:	SU004063
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Gender:	Male

Factors:

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

mb_sample_id	local_sample_id	Age	Condition	Sample source
SA445767	MAYO-01891	78 weeks	Control	Gastrocnemius Muscle
SA445768	MAYO-01888	78 weeks	Control	Gastrocnemius Muscle
SA445769	MAYO-01887	78 weeks	Control	Gastrocnemius Muscle
SA445770	MAYO-01890	78 weeks	Control	Gastrocnemius Muscle
SA445771	MAYO-01889	78 weeks	Control	Gastrocnemius Muscle
SA445772	MAYO-01892	78 weeks	KPC	Gastrocnemius Muscle
SA445773	MAYO-01893	78 weeks	KPC	Gastrocnemius Muscle
SA445774	MAYO-01894	78 weeks	KPC	Gastrocnemius Muscle
SA445775	MAYO-01895	78 weeks	KPC	Gastrocnemius Muscle
SA445776	MAYO-01896	78 weeks	KPC	Gastrocnemius Muscle
SA445777	MAYO-01897	78 weeks	KPC	Gastrocnemius Muscle
SA445778	MAYO-01898	78 weeks	KPC	Gastrocnemius Muscle
SA445779	MAYO-01899	78 weeks	KPC	Gastrocnemius Muscle
SA445780	MAYO-01904	8 weeks	Control	Gastrocnemius Muscle
SA445781	MAYO-01902	8 weeks	Control	Gastrocnemius Muscle
SA445782	MAYO-01903	8 weeks	Control	Gastrocnemius Muscle
SA445783	MAYO-01901	8 weeks	Control	Gastrocnemius Muscle
SA445784	MAYO-01900	8 weeks	Control	Gastrocnemius Muscle
SA445785	MAYO-01905	8 weeks	KPC	Gastrocnemius Muscle
SA445786	MAYO-01906	8 weeks	KPC	Gastrocnemius Muscle
SA445787	MAYO-01907	8 weeks	KPC	Gastrocnemius Muscle
SA445788	MAYO-01908	8 weeks	KPC	Gastrocnemius Muscle
SA445789	MAYO-01909	8 weeks	KPC	Gastrocnemius Muscle
SA445790	MAYO-01910	8 weeks	KPC	Gastrocnemius Muscle
SA445791	MAYO-01911	8 weeks	KPC	Gastrocnemius Muscle

Showing results 1 to 25 of 25

Showing results 1 to 25 of 25

Collection:

Collection ID:	CO004056
Collection Summary:	Mice were sacrificed at the survival endpoint. Survival endpoint was based on body score and/or tumor volume and/or body weight loss. Gastrocnemius muscles were then harvested and flash frozen. 40mg of frozen tissue was sent to Metabolon. All the mice in this study were included in this metabolomic database. KPC cells were obtained from Dr. Gina Razidlo (Mayo Clinic) and cultured in DMEM plus 10% FBS. Mice were orthotopically implanted with 5000 KPC cells.
Sample Type:	Gastrocnemius

Treatment:

Treatment ID:	TR004072
Treatment Summary:	Young (8 weeks) and Aged (78 weeks) mice were either injected with saline or KPC cells.

Sample Preparation:

Sampleprep ID:	SP004069
Sampleprep Summary:	Frozen gastrocnemius muscles were sent to Metabolon. These are the Metabolon Procedures: Sample Preparation: Samples were prepared using the automated MicroLab STAR® system from Hamilton Company. Several recovery standards were added prior to the first step in the extraction process for QC purposes. To remove protein, dissociate small molecules bound to protein or trapped in the precipitated protein matrix, and to recover chemically diverse metabolites, proteins were precipitated with methanol under vigorous shaking for 2 min (Glen Mills GenoGrinder 2000) followed by centrifugation. The resulting extract was divided into five fractions: two for analysis by two separate reverse phase (RP)/UPLC-MS/MS methods with positive ion mode electrospray ionization (ESI), one for analysis by RP/UPLC-MS/MS with negative ion mode ESI, one for analysis by HILIC/UPLC-MS/MS with negative ion mode ESI, and one sample was reserved for backup. Samples were placed briefly on a TurboVap® (Zymark) to remove the organic solvent. The sample extracts were stored overnight under nitrogen before preparation for analysis. Ultrahigh Performance Liquid Chromatography-Tandem Mass Spectroscopy (UPLC-MS/MS): All methods utilized a Waters ACQUITY ultra-performance liquid chromatography (UPLC) and a Thermo Scientific Q-Exactive high resolution/accurate mass spectrometer interfaced with a heated electrospray ionization (HESI-II) source and Orbitrap mass analyzer operated at 35,000 mass resolution. The sample extract was dried then reconstituted in solvents compatible to each of the four methods. Each reconstitution solvent contained a series of standards at fixed concentrations to ensure injection and chromatographic consistency. One aliquot was analyzed using acidic positive ion conditions, chromatographically optimized for more hydrophilic compounds. In this method, the extract was gradient eluted from a C18 column (Waters UPLC BEH C18-2.1x100 mm, 1.7 µm) using water and methanol, containing 0.05% perfluoropentanoic acid (PFPA) and 0.1% formic acid (FA). Another aliquot was also analyzed using acidic positive ion conditions, however it was chromatographically optimized for more hydrophobic compounds. In this method, the extract was gradient eluted from the same afore mentioned C18 column using methanol, acetonitrile, water, 0.05% PFPA and 0.01% FA and was operated at an overall higher organic content. Another aliquot was analyzed using basic negative ion optimized conditions using a separate dedicated C18 column. The basic extracts were gradient eluted from the column using methanol and water, however with 6.5mM Ammonium Bicarbonate at pH 8. The fourth aliquot was analyzed via negative ionization following elution from a HILIC column (Waters UPLC BEH Amide 2.1x150 mm, 1.7 µm) using a gradient consisting of water and acetonitrile with 10mM Ammonium Formate, pH 10.8. The MS analysis alternated between MS and data-dependent MSn scans using dynamic exclusion. The scan range varied slighted between methods but covered 70-1000 m/z.

Sampleprep ID:

SP004069

Sampleprep Summary:

Frozen gastrocnemius muscles were sent to Metabolon. These are the Metabolon Procedures: Sample Preparation: Samples were prepared using the automated MicroLab STAR® system from Hamilton Company. Several recovery standards were added prior to the first step in the extraction process for QC purposes. To remove protein, dissociate small molecules bound to protein or trapped in the precipitated protein matrix, and to recover chemically diverse metabolites, proteins were precipitated with methanol under vigorous shaking for 2 min (Glen Mills GenoGrinder 2000) followed by centrifugation. The resulting extract was divided into five fractions: two for analysis by two separate reverse phase (RP)/UPLC-MS/MS methods with positive ion mode electrospray ionization (ESI), one for analysis by RP/UPLC-MS/MS with negative ion mode ESI, one for analysis by HILIC/UPLC-MS/MS with negative ion mode ESI, and one sample was reserved for backup. Samples were placed briefly on a TurboVap® (Zymark) to remove the organic solvent. The sample extracts were stored overnight under nitrogen before preparation for analysis. Ultrahigh Performance Liquid Chromatography-Tandem Mass Spectroscopy (UPLC-MS/MS): All methods utilized a Waters ACQUITY ultra-performance liquid chromatography (UPLC) and a Thermo Scientific Q-Exactive high resolution/accurate mass spectrometer interfaced with a heated electrospray ionization (HESI-II) source and Orbitrap mass analyzer operated at 35,000 mass resolution. The sample extract was dried then reconstituted in solvents compatible to each of the four methods. Each reconstitution solvent contained a series of standards at fixed concentrations to ensure injection and chromatographic consistency. One aliquot was analyzed using acidic positive ion conditions, chromatographically optimized for more hydrophilic compounds. In this method, the extract was gradient eluted from a C18 column (Waters UPLC BEH C18-2.1x100 mm, 1.7 µm) using water and methanol, containing 0.05% perfluoropentanoic acid (PFPA) and 0.1% formic acid (FA). Another aliquot was also analyzed using acidic positive ion conditions, however it was chromatographically optimized for more hydrophobic compounds. In this method, the extract was gradient eluted from the same afore mentioned C18 column using methanol, acetonitrile, water, 0.05% PFPA and 0.01% FA and was operated at an overall higher organic content. Another aliquot was analyzed using basic negative ion optimized conditions using a separate dedicated C18 column. The basic extracts were gradient eluted from the column using methanol and water, however with 6.5mM Ammonium Bicarbonate at pH 8. The fourth aliquot was analyzed via negative ionization following elution from a HILIC column (Waters UPLC BEH Amide 2.1x150 mm, 1.7 µm) using a gradient consisting of water and acetonitrile with 10mM Ammonium Formate, pH 10.8. The MS analysis alternated between MS and data-dependent MSn scans using dynamic exclusion. The scan range varied slighted between methods but covered 70-1000 m/z.

Chromatography:

Chromatography ID:	CH004896
Chromatography Summary:	Low pH polar (LC/MS Pos early)
Instrument Name:	Waters Acquity
Column Name:	Waters Acquity BEH C18 (100 x 2.1mm, 1.7um)
Column Temperature:	40-50
Flow Gradient:	Linear gradient from 5% B to 80% B over 3.35 minutes
Flow Rate:	0.35 mL/min
Solvent A:	100% water; 0.1% formic acid; 0.05% PFPA, pH ~2.5
Solvent B:	100% methanol; 0.1% formic acid; 0.05% PFPA, pH ~2.5
Chromatography Type:	Reversed phase

Analysis:

Analysis ID:	AN006450
Analysis Type:	MS
Chromatography ID:	CH004896
Num Factors:	4
Num Metabolites:	847
Rt Units:	Seconds
Units:	raw area counts