Summary of Study ST003185

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 PR001983. The data can be accessed directly via it's Project DOI: 10.21228/M8D72S 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	ST003185
Study Title	A multimodal drug-diet-immunotherapy combination restrains melanoma progression and metastasis - plasma lipidomics
Study Summary	Lipidomics profiling of immunocompetent B16F10 model of melanoma to examine lipid levels in mouse plasma following reduced oleic acid content in diet combined with the stearyl CoA desaturase inhibitor CAY10566.
Institute	University of Colorado Anschutz Medical Campus
Last Name	Haines
First Name	Julie
Address	12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Email	julie.haines@cuanschutz.edu
Phone	3037243339
Submit Date	2024-05-02
Raw Data Available	Yes
Raw Data File Type(s)	raw(Thermo)
Analysis Type Detail	LC-MS
Release Date	2024-05-18
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR001983
Project DOI:	doi: 10.21228/M8D72S
Project Title:	A multimodal drug-diet-immunotherapy combination restrains melanoma progression and metastasis
Project Summary:	The genetic landscape of cancer cells can lead to specific metabolic dependencies for tumor growth. Dietary interventions represent an attractive strategy to restrict the availability of key nutrients to tumors. In this study, we identified that growth of a subset of melanoma was severely restricted by a rationally designed combination therapy of a stearoyl-CoA desaturase (SCD) inhibitor with an isocaloric low-oleic acid diet. Despite its importance in oncogenesis, SCD underwent monoallelic co-deletion along with PTEN on chromosome 10q in about 47.5% of melanoma, and the other SCD allele was methylated, resulting in very low SCD expression. While this SCD deficient subset was refractory to SCD inhibitors, the subset of PTEN wildtype melanoma that retained SCD was sensitive. As dietary oleic acid could potentially blunt the effect of SCD inhibitors, a low-oleic acid custom diet was combined with SCD inhibitor. The combination reduced monounsaturated fatty acids and increased saturated fatty acids, inducing robust apoptosis and growth suppression and inhibiting lung metastasis with minimal toxicity in preclinical mouse models of PTEN wildtype melanoma. When combined with anti-PD1 immunotherapy, the SCD inhibitor improved T cell functionality and further constrained melanoma growth in mice. Collectively, these results suggest that optimizing SCD inhibitors with diets low in oleic acid may offer a viable and efficacious therapeutic approach for improving melanoma treatment.
Institute:	University of Colorado Anschutz Medical Campus
Laboratory:	Lab of Angelo D'Alessandro in collaboration with lab of Biplab Dasgupta
Last Name:	Haines
First Name:	Julie
Address:	12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Email:	julie.haines@cuanschutz.edu
Phone:	3037243339

Subject:

Subject ID:	SU003304
Subject Type:	Mammal
Subject Species:	Mus musculus
Gender:	Not applicable

Factors:

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

mb_sample_id	local_sample_id	factor	Sample source
SA346787	DS2-039-039	SCD inhibitor + high oleic acid diet	plasma
SA346788	DS2-039-040	SCD inhibitor + high oleic acid diet	plasma
SA346789	DS2-039-041	SCD inhibitor + high oleic acid diet	plasma
SA346790	DS2-039-042	SCD inhibitor + high oleic acid diet	plasma
SA346791	DS2-039-044	SCD inhibitor + high oleic acid diet	plasma
SA346792	DS2-039-043	SCD inhibitor + high oleic acid diet	plasma
SA346793	DS2-039-032	SCD inhibitor + low oleic acid diet	plasma
SA346794	DS2-039-031	SCD inhibitor + low oleic acid diet	plasma
SA346795	DS2-039-028	SCD inhibitor + low oleic acid diet	plasma
SA346796	DS2-039-027	SCD inhibitor + low oleic acid diet	plasma
SA346797	DS2-039-030	SCD inhibitor + low oleic acid diet	plasma
SA346798	DS2-039-029	SCD inhibitor + low oleic acid diet	plasma
SA346799	DS2-039-038	Vehicle + high oleic acid diet	plasma
SA346800	DS2-039-037	Vehicle + high oleic acid diet	plasma
SA346801	DS2-039-033	Vehicle + high oleic acid diet	plasma
SA346802	DS2-039-036	Vehicle + high oleic acid diet	plasma
SA346803	DS2-039-034	Vehicle + high oleic acid diet	plasma
SA346804	DS2-039-035	Vehicle + high oleic acid diet	plasma
SA346805	DS2-039-023	vehicle + low oleic acid diet	plasma
SA346806	DS2-039-022	vehicle + low oleic acid diet	plasma
SA346807	DS2-039-024	vehicle + low oleic acid diet	plasma
SA346808	DS2-039-026	vehicle + low oleic acid diet	plasma
SA346809	DS2-039-021	vehicle + low oleic acid diet	plasma
SA346810	DS2-039-025	vehicle + low oleic acid diet	plasma

Showing results 1 to 24 of 24

Showing results 1 to 24 of 24

Collection:

Collection ID:	CO003297
Collection Summary:	After two weeks of treatment, retroorbital whole blood samples were collected following Isoflurane sedation. The samples were obtained using heparinized capillary tubes (Catalog Number: 22-362566). Subsequently, these samples were placed into tubes containing K2E (Catalog Number: 365974), with a bulb used to drain the capillary tube (Catalog Number: 22-362566). The tubes were centrifuged for 15 minutes at 2,000 x g (4°C). The resulting plasma (supernatant) was collected and aliquoted into 1.5ml Eppendorf tubes and stored at -80°C.
Sample Type:	Blood (plasma)

Treatment:

Treatment ID:	TR003313
Treatment Summary:	All animal procedures were carried out in accordance with the IACUC-approved protocol of Cincinnati Children’s Hospital Medical Center. Animals were monitored daily by Veterinary Services. For subcutaneous tumor implantation, 1x10^6 B16F10 cells were mixed with matrigel 1:1 and implanted subcutaneously into NOD-SCID IL2Rgnull or C57BL/6 mice, respectively. CAY10566 was suspended in 0.5% Methyl Cellulose in water with 0.2% Tween 80 at 9 parts per 1 part DMSO stock. CAY10566 was administered at 30mg/kg via oral gavage, twice daily, with a drug holiday during the weekends. Treatment began three days after tumor cell implantation (bilateral tumors in n=4 per group). Mice were euthanized17 days after tumor cell injection/14 days after the start of treatment.

Treatment ID:

TR003313

Treatment Summary:

All animal procedures were carried out in accordance with the IACUC-approved protocol of Cincinnati Children’s Hospital Medical Center. Animals were monitored daily by Veterinary Services. For subcutaneous tumor implantation, 1x10^6 B16F10 cells were mixed with matrigel 1:1 and implanted subcutaneously into NOD-SCID IL2Rgnull or C57BL/6 mice, respectively. CAY10566 was suspended in 0.5% Methyl Cellulose in water with 0.2% Tween 80 at 9 parts per 1 part DMSO stock. CAY10566 was administered at 30mg/kg via oral gavage, twice daily, with a drug holiday during the weekends. Treatment began three days after tumor cell implantation (bilateral tumors in n=4 per group). Mice were euthanized17 days after tumor cell injection/14 days after the start of treatment.

Sample Preparation:

Sampleprep ID:	SP003311
Sampleprep Summary:	For plasma analysis, 20 µL of plasma was aliquoted into 1.5 mL Eppendorf tubes followed by 480 µL cold MeOH. Samples were then vortexed for 30 minutes, insoluble material was pelleted by centrifugation (18000g, 10 min) and supernatants were isolated for analysis by UHPLC-MS.
Processing Storage Conditions:	4℃
Extract Storage:	-80℃

Combined analysis:

Analysis ID	AN005231	AN005232	AN005233
Analysis type	MS	MS	MS
Chromatography type	Reversed phase	Reversed phase	Reversed phase
Chromatography system	Thermo Vanquish	Thermo Vanquish	Thermo Vanquish
Column	Phenomenex Kinetex C18 (30 x 2.1mm,1.7um)	Phenomenex Kinetex C18 (30 x 2.1mm,1.7um)	Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um)
MS Type	ESI	ESI	ESI
MS instrument type	Orbitrap	Orbitrap	Orbitrap
MS instrument name	Thermo Q Exactive Orbitrap	Thermo Q Exactive Orbitrap	Thermo Q Exactive Orbitrap
Ion Mode	NEGATIVE	POSITIVE	NEGATIVE
Units	peak area	peak area	peak area

Chromatography:

Chromatography ID:	CH003958
Chromatography Summary:	Negative C18 global lipidomics
Instrument Name:	Thermo Vanquish
Column Name:	Phenomenex Kinetex C18 (30 x 2.1mm,1.7um)
Column Temperature:	45
Flow Gradient:	0-3 min 10-95% B at 0.3 mL/min, 3-4.2 min hold at 95% B at 0.3 mL/min, 4.2-4.3 min drop to 10% B at 0.45 mL/min, 4.3-4.9 min hold at 10%B while lowering flow rate to 0.4 mL/min, 4.9-5 min hold at 10%B while lowering flow rate to 0.3 mL/min.
Flow Rate:	0.3-0.4 ml/min
Sample Injection:	10 uL
Solvent A:	75% water/25% acetonitrile; 5 mM ammonium acetate
Solvent B:	90% isopropanol/10% acetonitrile; 5 mM ammonium acetate
Chromatography Type:	Reversed phase

Chromatography ID:	CH003959
Chromatography Summary:	Positive C18 global lipidomics
Instrument Name:	Thermo Vanquish
Column Name:	Phenomenex Kinetex C18 (30 x 2.1mm,1.7um)
Column Temperature:	45
Flow Gradient:	0-3 min 30-100% B at 0.3 mL/min, 3-4.2 min hold at 100% B at 0.3 mL/min, 4.2-4.3 min 100-30% B at 0.4 mL/min, 4.3-4.9 min hold at 30%B and 0.4 mL/min, 4.9-5 min hold at 30%B while lowering flow rate to 0.3 mL/min
Flow Rate:	0.3-0.4 ml/min
Sample Injection:	10 uL
Solvent A:	75% water/25% acetonitrile; 5 mM ammonium acetate
Solvent B:	90% isopropanol/10% acetonitrile; 5 mM ammonium acetate
Chromatography Type:	Reversed phase

Chromatography ID:	CH003960
Chromatography Summary:	Negative C18 fatty acids
Instrument Name:	Thermo Vanquish
Column Name:	Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um)
Column Temperature:	45
Flow Gradient:	0-1 min 25% B and 0.3 ml/min; 1-2 min 25-50% B and 0.3 mL/min; 2–8 min 50–90% B and 0.3 mL/min; 8–10 min 90–99% B and 0.3 mL/min; 10–14 min hold at 99% B and 0.3 mL/min; 14–14.1 min 99–25% B and 0.3 mL/min; 14.1–16.9 min hold at 25% B and 0.4 mL/min; 16.9–17 min hold at 25% B and resume flow of 0.3 mL/min
Flow Rate:	0.3-0.4 ml/min
Solvent A:	75% water/25% acetonitrile; 5 mM ammonium acetate
Solvent B:	50% isopropanol/45% acetonitrile/5% water; 5 mM ammonium acetate
Chromatography Type:	Reversed phase

MS:

MS ID:	MS004964
Analysis ID:	AN005231
Instrument Name:	Thermo Q Exactive Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	The Q Exactive was run independently in positive and negative ion mode, scanning using full MS from 125-1500 m/z at 70,000 resolution and top 10 data-dependent MS2 at 17,500 resolution. Electrospray ionization was achieved with 45 Arb sheath gas, 25 Arb auxiliary gas, and 4 kV spray voltage. Calibration was performed prior to the run using the PierceTM Positive and Negative Ion Calibration Solutions (Thermo Fisher Scientific). Run order of samples was randomized and technical replicates were injected after every 4 samples to assess quality control.
Ion Mode:	NEGATIVE

MS ID:	MS004965
Analysis ID:	AN005232
Instrument Name:	Thermo Q Exactive Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	The Q Exactive was run independently in positive and negative ion mode, scanning using full MS from 125-1500 m/z at 70,000 resolution and top 10 data-dependent MS2 at 17,500 resolution. Electrospray ionization was achieved with 45 Arb sheath gas, 25 Arb auxiliary gas, and 4 kV spray voltage. Calibration was performed prior to the run using the PierceTM Positive and Negative Ion Calibration Solutions (Thermo Fisher Scientific). Run order of samples was randomized and technical replicates were injected after every 4 samples to assess quality control.
Ion Mode:	POSITIVE

MS ID:	MS004966
Analysis ID:	AN005233
Instrument Name:	Thermo Q Exactive Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	The Q Exactive was run independently in positive and negative ion mode, scanning using full MS from 125-1500 m/z at 70,000 resolution and top 10 data-dependent MS2 at 17,500 resolution. Electrospray ionization was achieved with 45 Arb sheath gas, 25 Arb auxiliary gas, and 4 kV spray voltage. Calibration was performed prior to the run using the PierceTM Positive and Negative Ion Calibration Solutions (Thermo Fisher Scientific). Run order of samples was randomized and technical replicates were injected after every 4 samples to assess quality control.
Ion Mode:	NEGATIVE