#METABOLOMICS WORKBENCH Stopka28_20220119_144647 DATATRACK_ID:3047 STUDY_ID:ST002072 ANALYSIS_ID:AN003377 PROJECT_ID:PR001314 VERSION 1 CREATED_ON January 31, 2022, 2:28 pm #PROJECT PR:PROJECT_TITLE A non-dividing population with high pyruvate dehydrogenase kinase activity PR:PROJECT_TITLE drives metabolic heterogeneity and tumorigenesis in the intestine PR:PROJECT_SUMMARY Although reprogramming of cellular metabolism is a hallmark of cancer, little is PR:PROJECT_SUMMARY known about how metabolic reprogramming contributes to early stages of PR:PROJECT_SUMMARY transformation. Here, we show that the histone deacetylase SIRT6 regulates tumor PR:PROJECT_SUMMARY initiation during intestinal cancer by controlling glucose metabolism. Loss of PR:PROJECT_SUMMARY SIRT6 results in increased number of intestinal stem cells (ISCs), which PR:PROJECT_SUMMARY translates into enhanced tumor initiating potential in APCmin mice. More PR:PROJECT_SUMMARY importantly, we found a metabolic compartmentalization within the intestinal PR:PROJECT_SUMMARY epithelium and adenomas, where a rare population of cells exhibit features of PR:PROJECT_SUMMARY Warburg-like metabolism characterized by high pyruvate dehydrogenase kinase PR:PROJECT_SUMMARY (PDK) activity. Our results show that these cells are quiescent cells expressing PR:PROJECT_SUMMARY +4 ISCs and enteroendocrine markers. Active glycolysis in these cells suppresses PR:PROJECT_SUMMARY ROS accumulation and enhances their stem cell and tumorigenic potential. Our PR:PROJECT_SUMMARY studies reveal that aerobic glycolysis represents a highly heterogeneous feature PR:PROJECT_SUMMARY of cancer, and more importantly, they indicate that this metabolic adaptation PR:PROJECT_SUMMARY occurs in non-dividing cells, suggesting a role for the Warburg effect beyond PR:PROJECT_SUMMARY biomass production in tumors. PR:INSTITUTE Massachusetts General Hospital PR:LAST_NAME Mostoslavsky PR:FIRST_NAME Raul PR:ADDRESS 55 Fruit Street Boston, MA 02114 PR:EMAIL rmostoslavsky@mgh.harvard.edu PR:PHONE 617-643-3146 #STUDY ST:STUDY_TITLE A non-dividing population with high pyruvate dehydrogenase kinase activity ST:STUDY_TITLE drives metabolic heterogeneity and tumorigenesis in the intestine ST:STUDY_SUMMARY Although reprogramming of cellular metabolism is a hallmark of cancer, little is ST:STUDY_SUMMARY known about how metabolic reprogramming contributes to early stages of ST:STUDY_SUMMARY transformation. Here, we show that the histone deacetylase SIRT6 regulates tumor ST:STUDY_SUMMARY initiation during intestinal cancer by controlling glucose metabolism. Loss of ST:STUDY_SUMMARY SIRT6 results in increased number of intestinal stem cells (ISCs), which ST:STUDY_SUMMARY translates into enhanced tumor initiating potential in APCmin mice. More ST:STUDY_SUMMARY importantly, we found a metabolic compartmentalization within the intestinal ST:STUDY_SUMMARY epithelium and adenomas, where a rare population of cells exhibit features of ST:STUDY_SUMMARY Warburg-like metabolism characterized by high pyruvate dehydrogenase kinase ST:STUDY_SUMMARY (PDK) activity. Our results show that these cells are quiescent cells expressing ST:STUDY_SUMMARY +4 ISCs and enteroendocrine markers. Active glycolysis in these cells suppresses ST:STUDY_SUMMARY ROS accumulation and enhances their stem cell and tumorigenic potential. Our ST:STUDY_SUMMARY studies reveal that aerobic glycolysis represents a highly heterogeneous feature ST:STUDY_SUMMARY of cancer, and more importantly, they indicate that this metabolic adaptation ST:STUDY_SUMMARY occurs in non-dividing cells, suggesting a role for the Warburg effect beyond ST:STUDY_SUMMARY biomass production in tumors. ST:INSTITUTE Massachusetts General Hospital ST:DEPARTMENT Brigham and Women's Hospital ST:LAST_NAME Mostoslavsky ST:FIRST_NAME Raul ST:ADDRESS 55 Fruit Street Boston, MA 02114 ST:EMAIL rmostoslavsky@mgh.harvard.edu ST:PHONE 5189653364 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #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 - adenoma_colon_HCl_1 Treatment:adenoma RAW_FILE_NAME=adenoma_colon_HCl_1-total ion count.imzML SUBJECT_SAMPLE_FACTORS - adenoma_dan_HCl_2 Treatment:adenoma RAW_FILE_NAME=adenoma_dan_HCl_2-total ion count.imzML SUBJECT_SAMPLE_FACTORS - adenoma_dan_HCl_3 Treatment:adenoma RAW_FILE_NAME=adenoma_dan_HCl_3-total ion count.imzML #COLLECTION CO:COLLECTION_SUMMARY As stated in the paper: "Colorectal tissue was stored at -80 °C until CO:COLLECTION_SUMMARY processing. Cryosections of the colorectal tissue, containing the adenoma, were CO:COLLECTION_SUMMARY taken at 10 µm thickness and were mounted on indium tin oxide (ITO) slides for CO:COLLECTION_SUMMARY MALDI MSI analysis. Serial sections were obtained for MALDI MSI and CO:COLLECTION_SUMMARY immunofluorescence microscopy using a pPDH antibody and DAPI staining. The CO:COLLECTION_SUMMARY cryosections used for immunofluorescence were 5 µm in thickness. Fluorescent CO:COLLECTION_SUMMARY microscopy images were acquired using a 40x objective (Zeiss Observer Z.1, CO:COLLECTION_SUMMARY Oberkochen, Germany), a DAPI filter (Filter Set 49, Carl Zeiss Microscopy, CO:COLLECTION_SUMMARY Oberkochen, Germany), and an FITC filter (31001, Chroma Technology Corporation, CO:COLLECTION_SUMMARY Bellows Falls, VT)." CO:SAMPLE_TYPE Colon #TREATMENT TR:TREATMENT_SUMMARY N/A #SAMPLEPREP SP:SAMPLEPREP_SUMMARY As stated in the paper: "4.4 mg/mL of 1,5-diaminonapthalene hydrochloride (CAS: SP:SAMPLEPREP_SUMMARY 2243-62-41, Sigma-Aldrich, Darnstadt, Germany) was dissolved in 4/4.5/0.5 HPLC SP:SAMPLEPREP_SUMMARY grade water/ethanol/1 M HCl (v/v/v). 28 The 10 µm thick tissue sections were SP:SAMPLEPREP_SUMMARY sprayed using a TM-sprayer (HTX Technologies, Chapel Hill, NC) in a four-pass SP:SAMPLEPREP_SUMMARY method. The parameters of the matrix application set in the TM-sprayer were as SP:SAMPLEPREP_SUMMARY follows: spray nozzle velocity (1200 mm/min), track spacing (2 mm), flow rate SP:SAMPLEPREP_SUMMARY (0.09 mL/min), spray nozzle temperature (75 °C), and nitrogen gas pressure (10 SP:SAMPLEPREP_SUMMARY psi)" #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE None (Direct infusion) CH:INSTRUMENT_NAME timsTOF fleX CH:COLUMN_NAME none #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Bruker timsTOF fleX MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE MALDI MS:ION_MODE NEGATIVE MS:MS_COMMENTS SCilS 2022a pro #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Da MS_METABOLITE_DATA_START Samples adenoma_colon_HCl_1 adenoma_dan_HCl_2 adenoma_dan_HCl_3 Factors Treatment:adenoma Treatment:adenoma Treatment:adenoma fumarate 70.9469147 97.916687 269.25971 Succinate 13.7846333 312.742577 476.288666 a-ketoglutarate 8.17143059 20.8496666 38.0566139 pentose phosphate 29.6945839 14.286932 76.3336105 hexose phosphate 400.424622 116.034538 258.387848 palmitoleic 97.7862244 40.3751869 68.404213 oleic 1341.27612 272.815277 299.874115 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name fumarate Succinate a-ketoglutarate pentose phosphate hexose phosphate palmitoleic oleic METABOLITES_END #END