#METABOLOMICS WORKBENCH bagchipuja_20240812_215714 DATATRACK_ID:5098 STUDY_ID:ST003479 ANALYSIS_ID:AN005714 PROJECT_ID:PR002137 VERSION 1 CREATED_ON September 18, 2024, 10:35 pm #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 #STUDY ST:STUDY_TITLE Fatty acid analysis of similarly sized MC38 tumors from control-diet- or ST:STUDY_TITLE high-fat-diet-fed C57BL/6 mice revealed an increase in tumoral oleic acid in ST:STUDY_TITLE high-fat-diet-fed mice. ST:STUDY_SUMMARY Similarly sized MC38 tumors from control-diet- and high-fat-diet-fed mice were ST:STUDY_SUMMARY subject to fatty acid analysis via GC-FID. These data revealed an increase in ST:STUDY_SUMMARY oleic acid in tumors from high-fat-diet-fed mice. Oleic acid was the only fatty ST:STUDY_SUMMARY acid to exhibit this trend. ST:INSTITUTE Stanford University ST:LAST_NAME Bagchi ST:FIRST_NAME Sreya ST:ADDRESS 3373 Hillview Avenue ST:EMAIL bagchipuja@gmail.com ST:PHONE 4086214773 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN C57BL/6J #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - 8 Sample source:MC38 murine whole tumor | Treatment:CD- different size tumor SUBJECT_SAMPLE_FACTORS - 9 Sample source:MC38 murine whole tumor | Treatment:CD- different size tumor SUBJECT_SAMPLE_FACTORS - 10 Sample source:MC38 murine whole tumor | Treatment:CD- different size tumor SUBJECT_SAMPLE_FACTORS - 11 Sample source:MC38 murine whole tumor | Treatment:CD- different size tumor SUBJECT_SAMPLE_FACTORS - 12 Sample source:MC38 murine whole tumor | Treatment:CD- different size tumor SUBJECT_SAMPLE_FACTORS - 13 Sample source:MC38 murine whole tumor | Treatment:CD- different size tumor SUBJECT_SAMPLE_FACTORS - 14 Sample source:MC38 murine whole tumor | Treatment:CD- different size tumor SUBJECT_SAMPLE_FACTORS - 15 Sample source:MC38 murine whole tumor | Treatment:CD- different size tumor SUBJECT_SAMPLE_FACTORS - 16 Sample source:MC38 murine whole tumor | Treatment:HFD- different size tumor SUBJECT_SAMPLE_FACTORS - 17 Sample source:MC38 murine whole tumor | Treatment:HFD- different size tumor SUBJECT_SAMPLE_FACTORS - 18 Sample source:MC38 murine whole tumor | Treatment:HFD- different size tumor SUBJECT_SAMPLE_FACTORS - 19 Sample source:MC38 murine whole tumor | Treatment:HFD- different size tumor SUBJECT_SAMPLE_FACTORS - 20 Sample source:MC38 murine whole tumor | Treatment:HFD- different size tumor SUBJECT_SAMPLE_FACTORS - 21 Sample source:MC38 murine whole tumor | Treatment:HFD- different size tumor SUBJECT_SAMPLE_FACTORS - 22 Sample source:MC38 murine whole tumor | Treatment:HFD- different size tumor SUBJECT_SAMPLE_FACTORS - 23 Sample source:MC38 murine whole tumor | Treatment:HFD- different size tumor SUBJECT_SAMPLE_FACTORS - 24 Sample source:MC38 murine whole tumor | Treatment:HFD- different size tumor #COLLECTION CO:COLLECTION_SUMMARY MC38 tumors were isolated from control and high-fat diet fed mice, 21 days post CO:COLLECTION_SUMMARY tumor implantation. CO:SAMPLE_TYPE Whole tumor #TREATMENT TR:TREATMENT_SUMMARY Mice were divided into two groups- control and high-fat diet fed. Mice were fed TR:TREATMENT_SUMMARY the respective diets for 2 months starting at the age of 2 months. Subsequently TR:TREATMENT_SUMMARY tumor cells were injected to induce tumor growth. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Whole tumors were isolated and immediately snap 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:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Agilent 6890N CH:COLUMN_NAME Agilent DB-23 (60m x 0.25mm, 0.25um) CH:SOLVENT_A 3 M methanolic hydrochloric acid CH:SOLVENT_B 100% hexane CH:FLOW_GRADIENT Temperature-programmed GC CH:FLOW_RATE 1.5 mL/min CH:COLUMN_TEMPERATURE 250 CH:METHODS_FILENAME MC38_fatty_acid_GC_protocol.pdf CH:INTERNAL_STANDARD pentadecanoic acid (15:0) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME N/A (FID detection was used) MS:INSTRUMENT_TYPE Other MS:MS_TYPE Other MS:ION_MODE UNSPECIFIED MS:MS_COMMENTS No MS involved (FID detection was used) #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS nmol/mg dry wt MS_METABOLITE_DATA_START Samples 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 Factors Sample source:MC38 murine whole tumor | Treatment:CD- different size tumor Sample source:MC38 murine whole tumor | Treatment:CD- different size tumor Sample source:MC38 murine whole tumor | Treatment:CD- different size tumor Sample source:MC38 murine whole tumor | Treatment:CD- different size tumor Sample source:MC38 murine whole tumor | Treatment:CD- different size tumor Sample source:MC38 murine whole tumor | Treatment:CD- different size tumor Sample source:MC38 murine whole tumor | Treatment:CD- different size tumor Sample source:MC38 murine whole tumor | Treatment:CD- different size tumor Sample source:MC38 murine whole tumor | Treatment:HFD- different size tumor Sample source:MC38 murine whole tumor | Treatment:HFD- different size tumor Sample source:MC38 murine whole tumor | Treatment:HFD- different size tumor Sample source:MC38 murine whole tumor | Treatment:HFD- different size tumor Sample source:MC38 murine whole tumor | Treatment:HFD- different size tumor Sample source:MC38 murine whole tumor | Treatment:HFD- different size tumor Sample source:MC38 murine whole tumor | Treatment:HFD- different size tumor Sample source:MC38 murine whole tumor | Treatment:HFD- different size tumor Sample source:MC38 murine whole tumor | Treatment:HFD- different size tumor myristic acid 24.62548874 19.19269784 22.64876095 26.72959037 22.86173372 20.85157798 24.10481138 24.84394848 24.7505992 24.55050256 26.2227645 23.31811557 20.60746339 27.41597048 24.21730271 21.82329024 27.62547877 palmitic acid 312.5381804 411.9803791 429.7922397 396.7561757 394.6359224 321.512836 394.589811 436.5180004 403.0308803 439.7675326 410.3995737 431.7342589 441.1197289 464.3652301 327.6686553 362.2772351 355.8869637 palmitoleic acid 100.2126409 68.01028818 100.0109638 77.89948449 74.71652074 72.06262905 81.88799148 114.4767765 69.74511125 94.33625999 86.93115055 51.85280998 72.58391362 94.71152766 56.36161573 49.92230669 77.87629113 stearic acid 341.2002405 393.5018705 303.8725239 354.0281432 347.2674568 307.5214606 371.2759306 246.1368191 276.5824903 192.6709036 392.4677593 340.3802824 319.4037661 234.649432 240.2969335 280.8413768 262.2607409 oleic acid 535.2850682 416.5974067 523.4729789 388.0222689 465.8115923 369.8202996 478.5782565 705.3204791 621.7540823 1017.760646 739.2125457 657.6913766 651.8204991 836.9621374 453.7821614 419.9094357 630.2115295 linoleic acid 162.1101046 209.2588289 373.6311104 253.0533659 216.7107636 180.6704591 232.4993542 586.9551394 256.048235 443.8435319 265.6299951 281.2878432 297.3897434 355.6604731 188.8606438 266.8030961 257.2112956 alpha-linolenic acid 2.169789519 2.853464523 12.764433 4.344708276 4.060539906 2.847437278 3.579343165 20.38400586 6.036223112 13.81493 4.141995175 5.277519668 6.206365175 8.694367123 3.778035724 5.434709085 5.145024353 eicosenoic acid 8.657981749 9.077362678 8.332580569 6.139050113 7.109050644 5.417581354 7.212243254 11.26496731 9.049719841 10.75940135 12.19127773 10.90242269 9.18086385 10.62969899 6.513759127 6.772780868 6.972900763 Icosadienoic acid 35.34126637 13.65392076 10.10634195 11.51917341 8.439857417 9.326847934 10.91087724 10.12685217 8.507256838 9.887566646 9.147401439 10.72694995 12.45858848 10.84093186 6.482734561 11.88246029 6.934194709 dihomo-gamma-linolenic acid 15.49469027 20.65109583 20.39259298 25.61613705 24.06777077 35.03238579 34.47234093 16.88996386 12.03828587 7.505941796 15.20539245 13.25499545 11.71840851 10.21039426 8.988832495 19.78591252 8.13365434 arachidonic acid 165.6310784 305.043318 227.6951996 246.4008517 229.0766472 173.3921131 178.9580437 148.1466811 213.6216965 86.96542443 209.9998681 219.8378422 238.0248693 154.6142985 159.9869144 206.0861227 160.1721738 behenic acid 15.22335258 12.48508768 8.300739715 10.25393262 11.52242489 9.856679954 10.57374658 6.347736398 8.065675234 3.411134371 15.12515686 11.15828274 6.834697305 4.254382482 7.397822644 5.837480323 6.182031995 tetracosanoic acid 14.87574511 18.47724421 11.95591399 13.71794998 17.26122872 12.74651644 16.70573061 9.482505251 13.61794786 6.023435983 22.61848291 18.12793606 11.56849717 6.683532537 10.83132209 10.51406004 11.20631188 nervonic acid 13.48003264 22.6447906 15.53187171 15.43955624 17.46018789 13.27714469 16.32766205 9.781177146 15.42708711 5.813307498 18.37919781 19.24383798 14.14881753 8.568815041 14.1439171 12.11583038 9.127932788 docosahexaenoic acid 42.31878117 97.28994626 77.67631731 85.85521526 71.16887881 52.65128616 44.16440897 48.14431537 67.75659797 21.87547524 48.19669951 70.98313564 72.06144414 43.57231976 41.83762284 74.93142894 39.01650937 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name RT PubChem ID KEGG ID myristic acid 7.544 11005 C06424 palmitic acid 9.637 985 C00249 palmitoleic acid 10.079 445638 C08362 stearic acid 12.151 5281 C01530 oleic acid 12.571 445639 C00712 linoleic acid 13.315 5280450 C01595 alpha-linolenic acid 14.238 5280934 C06427 eicosenoic acid 15.364 46234793 C16526 Icosadienoic acid 16.171 6439848 C16525 dihomo-gamma-linolenic acid 16.684 5280581 C03242 arachidonic acid 17.028 444899 C00219 behenic acid 17.901 8215 C08281 tetracosanoic acid 21.848 11197 C08320 nervonic acid 22.723 5281120 C08323 docosahexaenoic acid 23.005 45934466 C06429 METABOLITES_END #END