#METABOLOMICS WORKBENCH bagchipuja_20240812_153809 DATATRACK_ID:5097 STUDY_ID:ST003478 ANALYSIS_ID:AN005713 PROJECT_ID:PR002137 VERSION 1 CREATED_ON 09-19-2024 #PROJECT PR:PROJECT_TITLE Acid-sensing receptor, GPR65, on tumor macrophages drives accelerated tumor PR:PROJECT_TITLE growth in obesity PR:PROJECT_SUMMARY Multiple cancers, including colorectal cancer (CRC), are more frequent and often PR:PROJECT_SUMMARY more aggressive in obese individuals. Here, we show that macrophages accumulate PR:PROJECT_SUMMARY within tumors of obese CRC patients and in obese CRC mice and promote PR:PROJECT_SUMMARY accelerated tumor growth. These changes are initiated by oleic acid accumulation PR:PROJECT_SUMMARY and subsequent tumor cell-derived acid production, and driven by signaling PR:PROJECT_SUMMARY through GPR65, an acid-sensing receptor on CRC-associated macrophages. We PR:PROJECT_SUMMARY demonstrate a similar role for GPR65 in hepatocellular carcinoma (HCC) in obese PR:PROJECT_SUMMARY mice. Tumors in obese patients with CRC or HCC also exhibit increased GPR65 PR:PROJECT_SUMMARY expression, suggesting that the mechanism revealed here likely contributes to PR:PROJECT_SUMMARY tumor growth in a range of obesity-associated cancers and represents a potential PR:PROJECT_SUMMARY therapeutic target. PR:INSTITUTE Stanford University PR:LAST_NAME Bagchi PR:FIRST_NAME Sreya PR:ADDRESS 3373 Hillview Avenue PR:EMAIL sbagchi@stanford.edu PR:PHONE 4086214773 PR:DOI http://dx.doi.org/10.21228/M8CZ33 #STUDY ST:STUDY_TITLE Fatty acid analysis of differently sized MC38 tumors harvested 3 weeks post ST:STUDY_TITLE implantation from control-diet- or high-fat-diet-fed C57BL/6 mice revealed an ST:STUDY_TITLE increase in tumoral oleic acid in high-fat-diet-fed mice. ST:STUDY_SUMMARY MC38 tumors were harvested 3 weeks post implantation from control-diet- and ST:STUDY_SUMMARY high-fat-diet-fed mice. At this point, tumors from high-fat-diet-fed mice were ST:STUDY_SUMMARY larger than that of control-diet-fed mice. These tumors were subject to fatty ST:STUDY_SUMMARY acid analysis via GC-FID. The resulting data revealed an increase in oleic acid ST:STUDY_SUMMARY in tumors from high-fat-diet-fed mice. Oleic acid was the only fatty acid to ST:STUDY_SUMMARY exhibit this trend. ST:INSTITUTE Stanford University ST:LAST_NAME Bagchi ST:FIRST_NAME Sreya ST:ADDRESS 3373 Hillview Avenue ST:EMAIL sbagchi@stanford.edu ST:PHONE 4086214773 ST:SUBMIT_DATE 2024-09-19 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN C57BL/6 SU:AGE_OR_AGE_RANGE 4-5 months #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - 1 Sample source:MC38 murine whole tumor | Treatment:CD- same size tumor SUBJECT_SAMPLE_FACTORS - 2 Sample source:MC38 murine whole tumor | Treatment:CD- same size tumor SUBJECT_SAMPLE_FACTORS - 3 Sample source:MC38 murine whole tumor | Treatment:CD- same size tumor SUBJECT_SAMPLE_FACTORS - 4 Sample source:MC38 murine whole tumor | Treatment:HFD- same size tumor SUBJECT_SAMPLE_FACTORS - 5 Sample source:MC38 murine whole tumor | Treatment:HFD- same size tumor SUBJECT_SAMPLE_FACTORS - 6 Sample source:MC38 murine whole tumor | Treatment:HFD- same size tumor SUBJECT_SAMPLE_FACTORS - 7 Sample source:MC38 murine whole tumor | Treatment:HFD- same size tumor #COLLECTION CO:COLLECTION_SUMMARY MC38 tumors from control and high-fat diet fed mice were collected 7-10 days CO:COLLECTION_SUMMARY post tumor implantation. CO:SAMPLE_TYPE Whole tumor #TREATMENT TR:TREATMENT_SUMMARY C57BL/6 mice were divided into two groups- control diet and high-fat diet. 2 TR:TREATMENT_SUMMARY month old mice were fed respective diets from 2 months, at which point MC38 TR:TREATMENT_SUMMARY tumors were implanted. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Whole tumors were resected and immediately frozen and sent for fatty acid SP:SAMPLEPREP_SUMMARY analysis. SP:SAMPLEPREP_PROTOCOL_FILENAME MC38_fatty_acid_GC_protocol.pdf #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY GC-FID was performed on an Agilent 6890N GC coupled to a FID ( flame ionization CH:CHROMATOGRAPHY_SUMMARY detector). The FID detector was operated at 260 °C. The hydrogen flow to the CH:CHROMATOGRAPHY_SUMMARY detector was 30 mL/min and air flow was 400 mL/min. The sampling rate of the FID CH:CHROMATOGRAPHY_SUMMARY was 20 Hz. CH:METHODS_FILENAME MC38_fatty_acid_GC_protocol.pdf CH:INSTRUMENT_NAME Agilent 6890N CH:COLUMN_NAME Agilent DB-23 (60m x 0.25mm, 0.25um) CH:COLUMN_TEMPERATURE 250 CH:FLOW_GRADIENT Temperature-programmed GC CH:FLOW_RATE 1.5 mL/min CH:INTERNAL_STANDARD pentadecanoic acid (15:0) CH:SOLVENT_A 3 M methanolic hydrochloric acid CH:SOLVENT_B 100% hexane CH:CHROMATOGRAPHY_TYPE GC #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME N/A (FID detection was used) MS:INSTRUMENT_TYPE Other MS:MS_TYPE Other MS:MS_COMMENTS No MS involved (FID detection was used) MS:ION_MODE UNSPECIFIED #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS nmol/mg dry wt MS_METABOLITE_DATA_START Samples 1 2 3 4 5 6 7 Factors Sample source:MC38 murine whole tumor | Treatment:CD- same size tumor Sample source:MC38 murine whole tumor | Treatment:CD- same size tumor Sample source:MC38 murine whole tumor | Treatment:CD- same size tumor Sample source:MC38 murine whole tumor | Treatment:HFD- same size tumor Sample source:MC38 murine whole tumor | Treatment:HFD- same size tumor Sample source:MC38 murine whole tumor | Treatment:HFD- same size tumor Sample source:MC38 murine whole tumor | Treatment:HFD- same size tumor alpha-linolenic acid 21.9680 21.9240 37.2880 22.1710 16.7120 17.7530 13.7210 arachidonic acid 61.5000 62.7590 45.9220 102.0880 50.4240 53.1720 67.9930 behenic acid 3.0300 2.2700 1.7330 7.5430 1.5470 1.7340 1.9020 dihomo-gamma-linolenic acid 6.4590 6.7120 6.7570 16.7590 5.8680 7.6960 7.3950 docosahexaenoic acid 22.5040 25.1910 23.9350 12.4120 11.3430 11.9230 15.9450 eicosenoic acid 16.1020 11.3540 8.7020 34.2370 11.1530 11.3490 14.7950 Icosadienoic acid 7.2930 5.0820 5.7140 28.0620 9.1310 8.7590 10.8400 linoleic acid 675.3100 596.6140 726.2310 936.7720 492.9790 501.9060 483.6820 myristic acid 26.6430 25.1590 31.4000 17.7910 22.4600 19.4530 24.0840 nervonic acid 6.3260 2.6930 3.0220 11.7810 2.0990 2.4260 3.3910 oleic acid 744.4500 724.6450 730.6600 2296.0560 1094.3140 1053.6140 1158.6790 palmitic acid 449.4670 427.2610 483.5580 962.3510 504.8740 473.9720 528.5140 palmitoleic acid 84.7340 125.7210 130.9970 151.5990 124.1490 110.0570 116.3980 stearic acid 160.9630 151.8430 116.2850 537.8660 144.0670 149.1650 189.2080 tetracosanoic acid 3.9330 3.8600 2.0010 7.1170 2.1070 1.9450 3.1950 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name pubchem_id inchi_key kegg_id other_id other_id_type ri ri_type moverz_quant alpha-linolenic acid 5280934 C06427 14.238 arachidonic acid 444899 C00219 17.028 behenic acid 8215 C08281 17.901 dihomo-gamma-linolenic acid 5280581 C03242 16.684 docosahexaenoic acid 45934466 C06429 23.005 eicosenoic acid 46234793 C16526 15.364 Icosadienoic acid 6439848 C16525 16.171 linoleic acid 5280450 C01595 13.315 myristic acid 11005 C06424 7.544 nervonic acid 5281120 C08323 22.723 oleic acid 445639 C00712 12.571 palmitic acid 985 C00249 9.637 palmitoleic acid 445638 C08362 10.079 stearic acid 5281 C01530 12.151 tetracosanoic acid 11197 C08320 21.848 METABOLITES_END #END