{
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"PROJECT":{"PROJECT_TITLE":"Mayo Pilot and Feasibility: H3K27M cells and glutamine metabolomics quatitation studies","PROJECT_SUMMARY":"In children, tumors affecting the brain and nervous system result in more cancer-related deaths than any other type of tumor. It is thus critical to identify new approaches for therapy. Among pediatric patients, one of the most devastating brain tumor types is Diffuse Intrinsic Pontine Gliomas (DIPG). Our understanding of this deadly disease has recently been advanced by important discoveries, including the discovery that the majority of DIPG tumors harbor the histone H3K27M mutation. This mutation results in global hypomethylation of H3K27 residues and is the pathological hallmark for this disease. Glutamine (Gln) addiction has been reported in many cancers including malignant adult gliomas. Glutamine likely promotes cancer cell proliferation and survival likely through generation of the TCA cycle intermediate alpha-ketoglutarate (α-KG). Importantly, α-KG is a critical co-factor for histone lysine demethylases including JMJD3, the enzyme responsible for removing the methyl groups from H3K27me3. Our preliminary data shows H3K27M tumor cells require Gln for survival, and if Gln is removed from the culture media, cells can be rescued by the addition of α-KG. Furthermore, Gln deprivation leads to an increase in H3K27 trimethylation similar to direct inhibition of JMJD3. It is for these reasons we hypothesize that H3K27M tumors are dependent on Gln derived α-KG both for feeding the TCA cycle and for further decreasing H3K27 trimethylation. Inhibition of Gln metabolism will likely uncover novel therapeutic targets for this deadly disease. In Aim 1 we will study Gln and glucose metabolism in H3K27M tumor cells and compare this to Wild Type (WT) tumors and Embryonic Stem Cells (ESCs). In Aim 2 we will validate the therapeutic validity of inhibiting Gln metabolism in H3K27M tumors.","INSTITUTE":"Mayo Clinic","LAST_NAME":"Daniels","FIRST_NAME":"David","ADDRESS":"200 First Street SW Rochester, MN 55905","EMAIL":"daniels.david@mayo.edu","PHONE":"507-284-2511"},

"STUDY":{"STUDY_TITLE":"TCA Isotopmer metabolomics of H3K27M Cells grown in regular media, glutamine enriched regular media, and glucose encriched regular media (Part-III)","STUDY_SUMMARY":"TCA Isotopmer metabolomics of H3K27M Cells grown in regular media, glutamine enriched regular media, and glucose encriched regular media. TCA isotopomers traced for 0, 12, or 24 hours were measured.","INSTITUTE":"Mayo Clinic","LAST_NAME":"Daniels","FIRST_NAME":"David","ADDRESS":"200 First Street SW Rochester, MN 55905","EMAIL":"daniels.david@mayo.edu","PHONE":"507-284-2511"},

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"COLLECTION":{"COLLECTION_SUMMARY":"DIPG IV and DIPG XVII cell lines are collected in this experiment. Susupension cells are harvested using centrifugation at 1200 rpm for 5 min. 1 mL of the supernatant media werr collected in an eppendorf tube and snapped frozen. The cell pellets were broken up into single cell suspension and counted. 1 million cells were taken from the stock and washed 1 x with PBS using table top centrifuge with 10 sec quick spin. The resulting cell pellet was snap frozen. Both the frozen media and cell pellet are stored in -80 C prior transfer.","SAMPLE_TYPE":"Glioma cells"},

"TREATMENT":{"TREATMENT_SUMMARY":"TCA isotopomer in H3K27M cell lines traced for 0, 12, or 24 hours. Cells grow in regular media without isotopes Cells grow in regular media enriched with 35% U-13C-glutamine Cells grow in regular media enriched with 35% U-13C-glucose Study Design Factors IV Basal - DIPG IV Regular Media No Tracers IV 12h GLN - DIPG IV Gultamine enriched media 12 hour Tracers IV 24h GLN - DIPG IV Gultamine enriched media 24 hour Tracers IV 12h GLC - DIPG IV Glucose enriched media 12 hour Tracers IV 24h GLC - DIPG IV Glucose enriched media 24 hour Tracers XVII Basal - DIPG XVII Regular Media No Tracers XVII 12h GLN - DIPG XVII Gultamine enriched media 12 hour Tracers XVII 24h GLN - DIPG XVII Gultamine enriched media 24 hour Tracers XVII 12h GLC - DIPG XVII Glucose enriched media 12 hour Tracers XVII 24h GLC - DIPG XVII Glucose enriched media 24 hour Tracers"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"TCA Isotopomers in glioma cell lines traced for 0, 12, 24 hours"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"GC","INSTRUMENT_NAME":"Agilent 7890B","COLUMN_NAME":"Agilent HP5-MS (30m × 0.25mm, 0.25 um)"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Agilent 5977A","INSTRUMENT_TYPE":"Single quadrupole","MS_TYPE":"EI","ION_MODE":"POSITIVE"},

"MS_METABOLITE_DATA":{
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}

}