{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001838","ANALYSIS_ID":"AN002980","VERSION":"1","CREATED_ON":"June 24, 2021, 2:45 pm"},

"PROJECT":{"PROJECT_TITLE":"Reversing Epigenetic Gene Silencing to Overcome Immune Evasion in CNS Malignancies","PROJECT_SUMMARY":"Glioblastoma is an aggressive brain malignancy with a dismal prognosis. With emerging evidence that disproves the immune privileged environment in the brain, there is much interest in examining various immunotherapy strategies to treat these incurable cancers. Unfortunately, to date, clinical studies investigating immunotherapy regimens have not provided much evidence of efficacy, leading to questions about the suitability of immunotherapy strategies for these tumors. Inadequate inherent populations of lymphocytes in tumor (TILs) and limited trafficking of systemic circulating T cells into the central nervous system (CNS) likely contribute to the poor response to immunotherapy treatment for primary CNS cancers. This paucity of TILs is in concert with the finding of epigenetic silencing of genes that promote immune cell movement (chemotaxis) to the tumor. In this study we evaluated the ability of GSK126, a blood-brain barrier permeable small molecule inhibitor of EZH2, to reverse the epigenetic silencing of chemokines like CXCL9 and CXCL10. When combined with anti-PD-1 treatment, these IFN driven chemokines promote T cell infiltration, resulting in decreased tumor growth and enhanced survival in immunocompetent murine sub-cutaneous and intracranial tumor syngeneic models of GBM. Examination of the tumor micro-environment revealed that the decrease in tumor growth in the mice treated with the drug combination was accompanied by increased tumor CD8 T cell infiltration along with higher IFN expression. Additionally, a significant increase in CXCR3+ T cells in the draining lymph nodes was also found. Taken together, our data suggests that in glioblastoma, epigenetic modulation using GSK126 could improve current immunotherapy strategies by reversing the epigenetic changes that enable immune cell evasion leading to enhanced immune c-ll trafficking to the tumor.","INSTITUTE":"National Cancer Institute","DEPARTMENT":"Neuro-Oncology Branch","LABORATORY":"Cancer Metabolism","LAST_NAME":"Dowdy","FIRST_NAME":"Tyrone","ADDRESS":"37 convent dr, Bldg 37 rm 1142","EMAIL":"tyrone.dowdy@nih.gov","PHONE":"2407607066","CONTRIBUTORS":"Nivedita M. Ratnam1, Heather M. Sonnemann1, Stephen C. Frederico1, Huanwen Chen1, Marsha-Kay N.D. Hutchinson1, Tyrone Dowdy1, Caitlin M. Reid1, Jinkyu Jung1, Wei Zhang1, Hua Song1, Meili Zhang1, Dionne Davis1, Mioara Larion1, Amber J. Giles1 and Mark R. Gilbert1*."},

"STUDY":{"STUDY_TITLE":"Reversing Epigenetic Gene Silencing to Overcome Immune Evasion in CNS Malignancies","STUDY_SUMMARY":"Glioblastoma is an aggressive brain malignancy with a dismal prognosis. With emerging evidence that disproves the immune privileged environment in the brain, there is much interest in examining various immunotherapy strategies to treat these incurable cancers. Unfortunately, to date, clinical studies investigating immunotherapy regimens have not provided much evidence of efficacy, leading to questions about the suitability of immunotherapy strategies for these tumors. Inadequate inherent populations of lymphocytes in tumor (TILs) and limited trafficking of systemic circulating T cells into the central nervous system (CNS) likely contribute to the poor response to immunotherapy treatment for primary CNS cancers. This paucity of TILs is in concert with the finding of epigenetic silencing of genes that promote immune cell movement (chemotaxis) to the tumor. In this study we evaluated the ability of GSK126, a blood-brain barrier permeable small molecule inhibitor of EZH2, to reverse the epigenetic silencing of chemokines like CXCL9 and CXCL10. When combined with anti-PD-1 treatment, these IFN driven chemokines promote T cell infiltration, resulting in decreased tumor growth and enhanced survival in immunocompetent murine sub-cutaneous and intracranial tumor syngeneic models of GBM. Examination of the tumor micro-environment revealed that the decrease in tumor growth in the mice treated with the drug combination was accompanied by increased tumor CD8 T cell infiltration along with higher IFN expression. Additionally, a significant increase in CXCR3+ T cells in the draining lymph nodes was also found. Taken together, our data suggests that in glioblastoma, epigenetic modulation using GSK126 could improve current immunotherapy strategies by reversing the epigenetic changes that enable immune cell evasion leading to enhanced immune cell trafficking to the tumor.","INSTITUTE":"National Cancer Institute","DEPARTMENT":"Neuro-Oncology Branch","LABORATORY":"Cancer Metabolism","LAST_NAME":"Dowdy","FIRST_NAME":"Tyrone","ADDRESS":"37 convent dr, Bldg 37 rm 1142","EMAIL":"tyrone.dowdy@nih.gov","PHONE":"2407607066"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090","GENOTYPE_STRAIN":"albino C57BL/6","AGE_OR_AGE_RANGE":"6-8 weeks","GENDER":"Female","ANIMAL_HOUSING":"NCI-Bethesda Animal Facility","ANIMAL_ANIMAL_SUPPLIER":"Jackson Laboratories (Bar Harbor, ME)"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"blank",
"Sample ID":"Blank_S14.d",
"Factors":{"Treatment":"blank","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"blank","Mice Tumor SI#":"blank","sample  quantity)":"-","(units)":"blank","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S14.d"}
},
{
"Subject ID":"blank",
"Sample ID":"blank_S22.d",
"Factors":{"Treatment":"blank","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"blank","Mice Tumor SI#":"blank","sample  quantity)":"-","(units)":"blank","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S22.d"}
},
{
"Subject ID":"blank",
"Sample ID":"blank_Sr62.d",
"Factors":{"Treatment":"blank","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"blank","Mice Tumor SI#":"blank","sample  quantity)":"-","(units)":"blank","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr62blk.d"}
},
{
"Subject ID":"Intra1_BRN_g1_2h_mg19.335",
"Sample ID":"BRN_g1 _Intra1_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.0"},
"Additional sample data":{"Specimen Group":"BRN_g1","Mice Tumor SI#":"Intracranial1","sample  quantity)":"19.335","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr73.d"}
},
{
"Subject ID":"Intra2_BRN_g1_2h_mg19.835",
"Sample ID":"BRN_g1 _Intra2_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.0"},
"Additional sample data":{"Specimen Group":"BRN_g1","Mice Tumor SI#":"Intracranial2","sample  quantity)":"19.835","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr74.d"}
},
{
"Subject ID":"Intra3_BRN_g1_2h_mg20.96",
"Sample ID":"BRN_g1 _Intra3_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.0"},
"Additional sample data":{"Specimen Group":"BRN_g1","Mice Tumor SI#":"Intracranial3","sample  quantity)":"20.96","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr75.d"}
},
{
"Subject ID":"Intra4_BRN_g1_6h_mg25.175",
"Sample ID":"BRN_g1 _Intra4_6h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"6.0"},
"Additional sample data":{"Specimen Group":"BRN_g1","Mice Tumor SI#":"Intracranial4","sample  quantity)":"25.175","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr76.d"}
},
{
"Subject ID":"Intra5_BRN_g1_6h_mg16.81",
"Sample ID":"BRN_g1 _Intra5_6h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"6.0"},
"Additional sample data":{"Specimen Group":"BRN_g1","Mice Tumor SI#":"Intracranial5","sample  quantity)":"16.81","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr77.d"}
},
{
"Subject ID":"Intra7_BRN_g1_10h_mg18.28",
"Sample ID":"BRN_g1 _Intra7_10h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"10.0"},
"Additional sample data":{"Specimen Group":"BRN_g1","Mice Tumor SI#":"Intracranial7","sample  quantity)":"18.28","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr78.d"}
},
{
"Subject ID":"Intra8_BRN_g1_10h_mg16",
"Sample ID":"BRN_g1 _Intra8_10h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"10.0"},
"Additional sample data":{"Specimen Group":"BRN_g1","Mice Tumor SI#":"Intracranial8","sample  quantity)":"16","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr79.d"}
},
{
"Subject ID":"Intra10_BRN_g2_2h_mg22.06",
"Sample ID":"BRN_ctrl_g2_Intra10_2h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"BRN_g2","Mice Tumor SI#":"Intracranial10","sample  quantity)":"22.06","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr93.d"}
},
{
"Subject ID":"Intra11_BRN_g2_2h_mg16.44",
"Sample ID":"BRN_ctrl_g2_Intra11_2h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"BRN_g2","Mice Tumor SI#":"Intracranial11","sample  quantity)":"16.44","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr94.d"}
},
{
"Subject ID":"Intra12_BRN_g2_6h_mg18.26",
"Sample ID":"BRN_ctrl_g2_Intra12_6h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"BRN_g2","Mice Tumor SI#":"Intracranial12","sample  quantity)":"18.26","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr95.d"}
},
{
"Subject ID":"Intra13_BRN_g2_6h_mg18.35",
"Sample ID":"BRN_ctrl_g2_Intra13_6h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"BRN_g2","Mice Tumor SI#":"Intracranial13","sample  quantity)":"18.35","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr96.d"}
},
{
"Subject ID":"Intra14_BRN_g2_6h_mg16",
"Sample ID":"BRN_ctrl_g2_Intra14_6h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"BRN_g2","Mice Tumor SI#":"Intracranial14","sample  quantity)":"16","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr97.d"}
},
{
"Subject ID":"Intra15_BRN_g2_10h_mg21.2",
"Sample ID":"BRN_ctrl_g2_Intra15_10h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"BRN_g2","Mice Tumor SI#":"Intracranial15","sample  quantity)":"21.2","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr98.d"}
},
{
"Subject ID":"Intra16_BRN_g2_10h_mg18.6",
"Sample ID":"BRN_ctrl_g2_Intra16_10h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"BRN_g2","Mice Tumor SI#":"Intracranial16","sample  quantity)":"18.6","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr99.d"}
},
{
"Subject ID":"SubCu1_DLN_g1_2h_mg1.92",
"Sample ID":"SubCu1_DLN_g1_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.00"},
"Additional sample data":{"Specimen Group":"DLN_g1","Mice Tumor SI#":"SubCu1_DLN","sample  quantity)":"1.92","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_004.d"}
},
{
"Subject ID":"SubCu2_DLN_g1_2h_mg8.04",
"Sample ID":"SubCu2_DLN_g1_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.00"},
"Additional sample data":{"Specimen Group":"DLN_g1","Mice Tumor SI#":"SubCu2_DLN","sample  quantity)":"8.04","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_008.d"}
},
{
"Subject ID":"SubCu3_DLN_g1_2h_mg2.945",
"Sample ID":"SubCu3_DLN_g1_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.00"},
"Additional sample data":{"Specimen Group":"DLN_g1","Mice Tumor SI#":"SubCu3_DLN","sample  quantity)":"2.945","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_007.d"}
},
{
"Subject ID":"SubCu4_DLN_g1_6h_mg1.82",
"Sample ID":"SubCu4_DLN_g1_6h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"6.00"},
"Additional sample data":{"Specimen Group":"DLN_g1","Mice Tumor SI#":"SubCu4_DLN","sample  quantity)":"1.82","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_017.d"}
},
{
"Subject ID":"SubCu5_DLN_g1_6h_mg3.145",
"Sample ID":"SubCu5_DLN_g1_6h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"6.00"},
"Additional sample data":{"Specimen Group":"DLN_g1","Mice Tumor SI#":"SubCu5_DLN","sample  quantity)":"3.145","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_018.d"}
},
{
"Subject ID":"SubCu6_DLN_g1_6h_mg2.21",
"Sample ID":"SubCu6_DLN_g1_6h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"6.00"},
"Additional sample data":{"Specimen Group":"DLN_g1","Mice Tumor SI#":"SubCu6_DLN","sample  quantity)":"2.21","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_021.d"}
},
{
"Subject ID":"SubCu7_DLN_g1_10h_mg1.5",
"Sample ID":"SubCu7_DLN_g1_10",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"10"},
"Additional sample data":{"Specimen Group":"DLN_g1","Mice Tumor SI#":"SubCu7_DLN","sample  quantity)":"1.5","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_023.d"}
},
{
"Subject ID":"SubCu8_DLN_g1_10h_mg3",
"Sample ID":"SubCu8_DLN_g1_10",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"10"},
"Additional sample data":{"Specimen Group":"DLN_g1","Mice Tumor SI#":"SubCu8_DLN","sample  quantity)":"3","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_024.d"}
},
{
"Subject ID":"SubCu9_DLN_g2_2h_mg10.475",
"Sample ID":"SubCu9_DLN_ctrl_g2_2h_",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"DLN_g2","Mice Tumor SI#":"SubCu9_DLN","sample  quantity)":"10.475","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_065.d"}
},
{
"Subject ID":"SubCu10_DLN_g2_2h_mg3.07",
"Sample ID":"SubCu10_DLN_ctrl_g2_2h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"DLN_g2","Mice Tumor SI#":"SubCu10_DLN","sample  quantity)":"3.07","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_066.d"}
},
{
"Subject ID":"SubCu11_DLN_g2_2h_mg2.49",
"Sample ID":"SubCu11_DLN_ctrl_g2_2h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"DLN_g2","Mice Tumor SI#":"SubCu11_DLN","sample  quantity)":"2.49","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_068.d"}
},
{
"Subject ID":"SubCu12_DLN_g2_6h_mg4.505",
"Sample ID":"SubCu12_DLN_ctrl_g2_6h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"DLN_g2","Mice Tumor SI#":"SubCu12_DLN","sample  quantity)":"4.505","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_069.d"}
},
{
"Subject ID":"SubCu13_DLN_g2_6h_mg12.535",
"Sample ID":"SubCu13_DLN_ctrl_g2_6h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"DLN_g2","Mice Tumor SI#":"SubCu13_DLN","sample  quantity)":"2.535","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_072.d"}
},
{
"Subject ID":"SubCu15_DLN_g2_10h_mg6.26",
"Sample ID":"SubCu15_DLN_ctrl_g2_10h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"DLN_g2","Mice Tumor SI#":"SubCu15_DLN","sample  quantity)":"6.26","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_075.d"}
},
{
"Subject ID":"SubCu16_DLN_g2_10h_mg5.39",
"Sample ID":"SubCu16_DLN_ctrl_g2_10h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"DLN_g2","Mice Tumor SI#":"SubCu16_DLN","sample  quantity)":"5.39","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_078.d"}
},
{
"Subject ID":"Intra1_LLN_g1_2h_mg3.05",
"Sample ID":"LLN_g1 _Intra1_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.0"},
"Additional sample data":{"Specimen Group":"LLN_g1","Mice Tumor SI#":"Intracranial1","sample  quantity)":"3.05","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_L1004.d"}
},
{
"Subject ID":"Intra2_LLN_g1_2h_mg5",
"Sample ID":"LLN_g1 _Intra2_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.0"},
"Additional sample data":{"Specimen Group":"LLN_g1","Mice Tumor SI#":"Intracranial2","sample  quantity)":"5","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_L1005.d"}
},
{
"Subject ID":"Intra3_LLN_g1_2h_mg1.975",
"Sample ID":"LLN_g1 _Intra3_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.0"},
"Additional sample data":{"Specimen Group":"LLN_g1","Mice Tumor SI#":"Intracranial3","sample  quantity)":"1.975","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_L1006.d"}
},
{
"Subject ID":"Intra4_LLN_g1_6h_mg1.03",
"Sample ID":"LLN_g1 _Intra4_6h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"6.0"},
"Additional sample data":{"Specimen Group":"LLN_g1","Mice Tumor SI#":"Intracranial4","sample  quantity)":"1.03","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_L1007.d"}
},
{
"Subject ID":"Intra6_LLN_g1_6h_mg0.975",
"Sample ID":"LLN_g1 _Intra6_6h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"6.0"},
"Additional sample data":{"Specimen Group":"LLN_g1","Mice Tumor SI#":"Intracranial6","sample  quantity)":"0.975","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_L1008.d"}
},
{
"Subject ID":"Intra7_LLN_g1_10h_mg0.975",
"Sample ID":"LLN_g1 _Intra7_10h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"10.0"},
"Additional sample data":{"Specimen Group":"LLN_g1","Mice Tumor SI#":"Intracranial7","sample  quantity)":"0.975","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_L1009.d"}
},
{
"Subject ID":"Intra8_LLN_g1_10h_mg1.935",
"Sample ID":"LLN_g1 _Intra8_10h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"10.0"},
"Additional sample data":{"Specimen Group":"LLN_g1","Mice Tumor SI#":"Intracranial8","sample  quantity)":"1.935","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_L1010.d"}
},
{
"Subject ID":"Intra10_LLN_g2_2h_mg0.87",
"Sample ID":"LLN_ctrl_g2_Intra10_2h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"LLN_g2","Mice Tumor SI#":"Intracranial10","sample  quantity)":"0.87","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_L1013.d"}
},
{
"Subject ID":"Intra11_LLN_g2_2h_mg1.65",
"Sample ID":"LLN_ctrl_g2_Intra11_2h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"LLN_g2","Mice Tumor SI#":"Intracranial11","sample  quantity)":"1.65","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_L1014.d"}
},
{
"Subject ID":"Intra12_LLN_g2_6h_mg0.965",
"Sample ID":"LLN_ctrl_g2_Intra12_6h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"LLN_g2","Mice Tumor SI#":"Intracranial12","sample  quantity)":"0.965","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_L1015.d"}
},
{
"Subject ID":"Intra13_LLN_g2_6h_mg2.885",
"Sample ID":"LLN_ctrl_g2_Intra13_6h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"LLN_g2","Mice Tumor SI#":"Intracranial13","sample  quantity)":"2.885","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_L1016.d"}
},
{
"Subject ID":"Intra15_LLN_g2_10h_mg1.345",
"Sample ID":"LLN_ctrl_g2_Intra15_10h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"LLN_g2","Mice Tumor SI#":"Intracranial15","sample  quantity)":"1.345","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_L1018.d"}
},
{
"Subject ID":"Intra16_LLN_g2_10h_mg1.25",
"Sample ID":"LLN_ctrl_g2_Intra16_10h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"LLN_g2","Mice Tumor SI#":"Intracranial16","sample  quantity)":"1.25","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_L1019.d"}
},
{
"Subject ID":"Intra9_LLN_g2_2h_mg0.8",
"Sample ID":"LLN_ctrl_g2 _Intra9_2h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"LLN_g2","Mice Tumor SI#":"Intracranial9","sample  quantity)":"0.8","(units)":"mg","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_L1012.d"}
},
{
"Subject ID":"SubCu1_NLN_g1_2h_mg2.08",
"Sample ID":"SubCu1_NLN_g1_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.00"},
"Additional sample data":{"Specimen Group":"NLN_g1","Mice Tumor SI#":"SubCu1_NLN","sample  quantity)":"2.08","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_005.d"}
},
{
"Subject ID":"SubCu2_NLN_g1_2h_mg5.82",
"Sample ID":"SubCu2_NLN_g1_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.00"},
"Additional sample data":{"Specimen Group":"NLN_g1","Mice Tumor SI#":"SubCu2_NLN","sample  quantity)":"5.82","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_006.d"}
},
{
"Subject ID":"SubCu3_NLN_g1_2h_mg2.03",
"Sample ID":"SubCu3_NLN_g1_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.00"},
"Additional sample data":{"Specimen Group":"NLN_g1","Mice Tumor SI#":"SubCu3_NLN","sample  quantity)":"2.03","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_009.d"}
},
{
"Subject ID":"SubCu4_NLN_g1_6h_mg3.64",
"Sample ID":"SubCu4_NLN_g1_6h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"6.00"},
"Additional sample data":{"Specimen Group":"NLN_g1","Mice Tumor SI#":"SubCu4_NLN","sample  quantity)":"3.64","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_016.d"}
},
{
"Subject ID":"SubCu5_NLN_g1_6h_mg1.8",
"Sample ID":"SubCu5_NLN_g1_6h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"6.00"},
"Additional sample data":{"Specimen Group":"NLN_g1","Mice Tumor SI#":"SubCu5_NLN","sample  quantity)":"1.8","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_020.d"}
},
{
"Subject ID":"SubCu6_NLN_g1_6h_mg1.62",
"Sample ID":"SubCu6_NLN_g1_6h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"6.00"},
"Additional sample data":{"Specimen Group":"NLN_g1","Mice Tumor SI#":"SubCu6_NLN","sample  quantity)":"1.62","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_019.d"}
},
{
"Subject ID":"SubCu7_NLN_g1_10h_mg2.36",
"Sample ID":"SubCu7_NLN_g1_10",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"10"},
"Additional sample data":{"Specimen Group":"NLN_g1","Mice Tumor SI#":"SubCu7_NLN","sample  quantity)":"2.36","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_022.d"}
},
{
"Subject ID":"SubCu8_NLN_g1_10h_mg3.48",
"Sample ID":"SubCu8_NLN_g1_10",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"10"},
"Additional sample data":{"Specimen Group":"NLN_g1","Mice Tumor SI#":"SubCu8_NLN","sample  quantity)":"3.48","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_025.d"}
},
{
"Subject ID":"SubCu9_NLN_g2_2h_mg4.45",
"Sample ID":"SubCu9_NLN_ctrl_g2_2h_",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"NLN_g2","Mice Tumor SI#":"SubCu9_NLN","sample  quantity)":"4.45","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_067.d"}
},
{
"Subject ID":"SubCu10_NLN_g2_2h_mg2.58",
"Sample ID":"SubCu10_NLN_ctrl_g2_2h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"NLN_g2","Mice Tumor SI#":"SubCu10_NLN","sample  quantity)":"2.58","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_063.d"}
},
{
"Subject ID":"SubCu11_NLN_g2_2h_mg4.5",
"Sample ID":"SubCu11_NLN_ctrl_g2_2h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"NLN_g2","Mice Tumor SI#":"SubCu11_NLN","sample  quantity)":"4.5","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_064.d"}
},
{
"Subject ID":"SubCu12_NLN_g2_6h_mg7.29",
"Sample ID":"SubCu12_NLN_ctrl_g2_6h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"NLN_g2","Mice Tumor SI#":"SubCu12_NLN","sample  quantity)":"7.29","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_070.d"}
},
{
"Subject ID":"SubCu13_NLN_g2_6h_mg3.37",
"Sample ID":"SubCu13_NLN_ctrl_g2_6h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"NLN_g2","Mice Tumor SI#":"SubCu13_NLN","sample  quantity)":"3.37","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_071.d"}
},
{
"Subject ID":"SubCu14_NLN_g2_6h_mg8.03",
"Sample ID":"SubCu14_NLN_ctrl_g2_6h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"NLN_g2","Mice Tumor SI#":"SubCu14_NLN","sample  quantity)":"8.03","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_074.d"}
},
{
"Subject ID":"SubCu15_NLN_g2_10h_mg2.81",
"Sample ID":"SubCu15_NLN_ctrl_g2_10h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"NLN_g2","Mice Tumor SI#":"SubCu15_NLN","sample  quantity)":"2.81","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_076.d"}
},
{
"Subject ID":"SubCu16_NLN_g2_10h_mg3.16",
"Sample ID":"SubCu16_NLN_ctrl_g2_10h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"NLN_g2","Mice Tumor SI#":"SubCu16_NLN","sample  quantity)":"3.16","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_077.d"}
},
{
"Subject ID":"QcBRN",
"Sample ID":"QcBRN_S45.d",
"Factors":{"Treatment":"QC","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"QcBRN","Mice Tumor SI#":"QcBRN","sample  quantity)":"-","(units)":"Qc","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S45.d"}
},
{
"Subject ID":"QcBRN",
"Sample ID":"QcBRN_Sr91.d",
"Factors":{"Treatment":"QC","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"QcBRN","Mice Tumor SI#":"QcBRN","sample  quantity)":"-","(units)":"Qc","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr91.d"}
},
{
"Subject ID":"QcBRN",
"Sample ID":"QcBRN_Sr111.d",
"Factors":{"Treatment":"QC","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"QcBRN","Mice Tumor SI#":"QcBRN","sample  quantity)":"-","(units)":"Qc","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr111.d"}
},
{
"Subject ID":"QcBRN",
"Sample ID":"QcBRN_Sr65.d",
"Factors":{"Treatment":"QC","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"QcBRN","Mice Tumor SI#":"QcBRN","sample  quantity)":"-","(units)":"Qc","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr65.d"}
},
{
"Subject ID":"QcLLN",
"Sample ID":"QcLLN_L1002.d",
"Factors":{"Treatment":"QC","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"QcLLN_Intra","Mice Tumor SI#":"QcLLN","sample  quantity)":"-","(units)":"Qc","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_L1002.d"}
},
{
"Subject ID":"QCpool g1 D_NLN",
"Sample ID":"QCpool g1 D_NLN_subcu_055.d",
"Factors":{"Treatment":"QC","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"QCpool g1 D_NLN","Mice Tumor SI#":"QCpool g1 D_NLN","sample  quantity)":"-","(units)":"Qc","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_055.d"}
},
{
"Subject ID":"QCpool g1 D_NLN",
"Sample ID":"QCpool g1 D_NLN_subcu_081.d",
"Factors":{"Treatment":"QC","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"QCpool g1 D_NLN","Mice Tumor SI#":"QCpool g1 D_NLN","sample  quantity)":"-","(units)":"Qc","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_081.d"}
},
{
"Subject ID":"QCpool g1 D_NLN",
"Sample ID":"QCpool g1 D_NLN_subcu_135.d",
"Factors":{"Treatment":"QC","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"QCpool g1 D_NLN","Mice Tumor SI#":"QCpool g1 D_NLN","sample  quantity)":"-","(units)":"Qc","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_135.d"}
},
{
"Subject ID":"QCpool g1 D_NLN",
"Sample ID":"QCpool g1 D_NLN_subcu_012.d",
"Factors":{"Treatment":"QC","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"QCpool g1 D_NLN","Mice Tumor SI#":"QCpool g1 D_NLN","sample  quantity)":"-","(units)":"Qc","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_012.d"}
},
{
"Subject ID":"QCpool g1 D_NLN",
"Sample ID":"QCpool g1 D_NLN_subcu_028.d",
"Factors":{"Treatment":"QC","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"QCpool g1 D_NLN","Mice Tumor SI#":"QCpool g1 D_NLN","sample  quantity)":"-","(units)":"Qc","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_028.d"}
},
{
"Subject ID":"QCpool g1 D_NLN",
"Sample ID":"QCpool g1 D_NLN_subcu_037.d",
"Factors":{"Treatment":"QC","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"QCpool g1 D_NLN","Mice Tumor SI#":"QCpool g1 D_NLN","sample  quantity)":"-","(units)":"Qc","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_037.d"}
},
{
"Subject ID":"QCpool g1 SubTum",
"Sample ID":"QCpool g1 SubTum_038.d",
"Factors":{"Treatment":"QC","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"QCpool g1 SubTum","Mice Tumor SI#":"QCpool g1 SubTum","sample  quantity)":"-","(units)":"Qc","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_038.d"}
},
{
"Subject ID":"QCpool g1 SubTum",
"Sample ID":"QCpool g1 SubTum_056.d",
"Factors":{"Treatment":"QC","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"QCpool g1 SubTum","Mice Tumor SI#":"QCpool g1 SubTum","sample  quantity)":"-","(units)":"Qc","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_056.d"}
},
{
"Subject ID":"QCpool g1 SubTum",
"Sample ID":"QCpool g1 SubTum_082.d",
"Factors":{"Treatment":"QC","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"QCpool g1 SubTum","Mice Tumor SI#":"QCpool g1 SubTum","sample  quantity)":"-","(units)":"Qc","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_082.d"}
},
{
"Subject ID":"QCpool g1 SubTum",
"Sample ID":"QCpool g1 SubTum_136.d",
"Factors":{"Treatment":"QC","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"QCpool g1 SubTum","Mice Tumor SI#":"QCpool g1 SubTum","sample  quantity)":"-","(units)":"Qc","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_136.d"}
},
{
"Subject ID":"QcSer",
"Sample ID":"QcSer_S24.d",
"Factors":{"Treatment":"QC","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"QcSer","Mice Tumor SI#":"QcSer_","sample  quantity)":"-","(units)":"Qc","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S24.d"}
},
{
"Subject ID":"QcSer",
"Sample ID":"QcSer_S44.d",
"Factors":{"Treatment":"QC","Gsk Exposure Time (h)":"-"},
"Additional sample data":{"Specimen Group":"QcSer","Mice Tumor SI#":"QcSer","sample  quantity)":"-","(units)":"Qc","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S44.d"}
},
{
"Subject ID":"Subcutaneous1_SER_g1_2h_mL0.05",
"Sample ID":"SER_g1 _Subcutaneous1_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.0"},
"Additional sample data":{"Specimen Group":"SER_g1","Mice Tumor SI#":"Subcutaneous1","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S6.d"}
},
{
"Subject ID":"Subcutaneous2_SER_g1_2h_mL0.05",
"Sample ID":"SER_g1 _Subcutaneous2_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.0"},
"Additional sample data":{"Specimen Group":"SER_g1","Mice Tumor SI#":"Subcutaneous2","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S7.d"}
},
{
"Subject ID":"Subcutaneous3_SER_g1_2h_mL0.05",
"Sample ID":"SER_g1 _Subcutaneous3_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.0"},
"Additional sample data":{"Specimen Group":"SER_g1","Mice Tumor SI#":"Subcutaneous3","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S8.d"}
},
{
"Subject ID":"Subcutaneous4_SER_g1_6h_mL0.05",
"Sample ID":"SER_g1 _Subcutaneous4_6h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"6.0"},
"Additional sample data":{"Specimen Group":"SER_g1","Mice Tumor SI#":"Subcutaneous4","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S9.d"}
},
{
"Subject ID":"Subcutaneous5_SER_g1_6h_mL0.05",
"Sample ID":"SER_g1 _Subcutaneous5_6h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"6.0"},
"Additional sample data":{"Specimen Group":"SER_g1","Mice Tumor SI#":"Subcutaneous5","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S10.d"}
},
{
"Subject ID":"Subcutaneous6_SER_g1_6h_mL0.05",
"Sample ID":"SER_g1 _Subcutaneous6_6h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"6.0"},
"Additional sample data":{"Specimen Group":"SER_g1","Mice Tumor SI#":"Subcutaneous6","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S11.d"}
},
{
"Subject ID":"Subcutaneous7_SER_g1_10h_mL0.05",
"Sample ID":"SER_g1 _Subcutaneous7_10h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"10.0"},
"Additional sample data":{"Specimen Group":"SER_g1","Mice Tumor SI#":"Subcutaneous7","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S12.d"}
},
{
"Subject ID":"Subcutaneous8_SER_g1_10h_mL0.05",
"Sample ID":"SER_g1 _Subcutaneous8_10h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"10.0"},
"Additional sample data":{"Specimen Group":"SER_g1","Mice Tumor SI#":"Subcutaneous8","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S13.d"}
},
{
"Subject ID":"Intra1_SER_g1_2h_mL0.05",
"Sample ID":"SER_g1 _Intra1_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.0"},
"Additional sample data":{"Specimen Group":"SER_g1","Mice Tumor SI#":"Intracranial1","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S35.d"}
},
{
"Subject ID":"Intra2_SER_g1_2h_mL0.05",
"Sample ID":"SER_g1 _Intra2_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.0"},
"Additional sample data":{"Specimen Group":"SER_g1","Mice Tumor SI#":"Intracranial2","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S36.d"}
},
{
"Subject ID":"Intra3_SER_g1_2h_mL0.05",
"Sample ID":"SER_g1 _Intra3_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.0"},
"Additional sample data":{"Specimen Group":"SER_g1","Mice Tumor SI#":"Intracranial3","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S37.d"}
},
{
"Subject ID":"Intra4_SER_g1_6h_mL0.05",
"Sample ID":"SER_g1 _Intra4_6h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"6.0"},
"Additional sample data":{"Specimen Group":"SER_g1","Mice Tumor SI#":"Intracranial4","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S38.d"}
},
{
"Subject ID":"Intra7_SER_g1_10h_mL0.05",
"Sample ID":"SER_g1 _Intra7_10h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"10.0"},
"Additional sample data":{"Specimen Group":"SER_g1","Mice Tumor SI#":"Intracranial7","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S41.d"}
},
{
"Subject ID":"Intra8_SER_g1_10h_mL0.05",
"Sample ID":"SER_g1 _Intra8_10h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"10.0"},
"Additional sample data":{"Specimen Group":"SER_g1","Mice Tumor SI#":"Intracranial8","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S42s.d"}
},
{
"Subject ID":"Intra5_SER_g1_6h_mL0.05",
"Sample ID":"SER_g1 _Intra5_6h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"6.0"},
"Additional sample data":{"Specimen Group":"SER_g1","Mice Tumor SI#":"Intracranial5","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr69.d"}
},
{
"Subject ID":"Intra6_SER_g1_6h_mL0.05",
"Sample ID":"SER_g1 _Intra6_6h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"6.0"},
"Additional sample data":{"Specimen Group":"SER_g1","Mice Tumor SI#":"Intracranial6","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_Sr70.d"}
},
{
"Subject ID":"Subcutaneous10_SER_g2_2h_mL0.05",
"Sample ID":"SER_ctrl_g2_Subcutaneous10_2h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"SER_g2","Mice Tumor SI#":"Subcutaneous10","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S16.d"}
},
{
"Subject ID":"Subcutaneous12_SER_g2_6h_mL0.05",
"Sample ID":"SER_ctrl_g2_Subcutaneous12_6h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"SER_g2","Mice Tumor SI#":"Subcutaneous12","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S17.d"}
},
{
"Subject ID":"Subcutaneous13_SER_g2_6h_mL0.05",
"Sample ID":"SER_ctrl_g2_Subcutaneous13_6h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"SER_g2","Mice Tumor SI#":"Subcutaneous13","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S18.d"}
},
{
"Subject ID":"Subcutaneous14_SER_g2_6h_mL0.05",
"Sample ID":"SER_ctrl_g2_Subcutaneous14_6h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"SER_g2","Mice Tumor SI#":"Subcutaneous14","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S19.d"}
},
{
"Subject ID":"Subcutaneous15_SER_g2_10h_mL0.05",
"Sample ID":"SER_ctrl_g2_Subcutaneous15_10h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"SER_g2","Mice Tumor SI#":"Subcutaneous15","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S20.d"}
},
{
"Subject ID":"Subcutaneous16_SER_g2_10h_mL0.05",
"Sample ID":"SER_ctrl_g2_Subcutaneous16_10h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"SER_g2","Mice Tumor SI#":"Subcutaneous16","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S21.d"}
},
{
"Subject ID":"Intra10_SER_g2_2h_mL0.05",
"Sample ID":"SER_ctrl_g2_Intra10_2h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"SER_g2","Mice Tumor SI#":"Intracranial10","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S27.d"}
},
{
"Subject ID":"Intra11_SER_g2_2h_mL0.05",
"Sample ID":"SER_ctrl_g2_Intra11_2h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"SER_g2","Mice Tumor SI#":"Intracranial11","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S28.d"}
},
{
"Subject ID":"Intra12_SER_g2_6h_mL0.05",
"Sample ID":"SER_ctrl_g2_Intra12_6h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"SER_g2","Mice Tumor SI#":"Intracranial12","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S29.d"}
},
{
"Subject ID":"Intra13_SER_g2_6h_mL0.05",
"Sample ID":"SER_ctrl_g2_Intra13_6h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"SER_g2","Mice Tumor SI#":"Intracranial13","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S30.d"}
},
{
"Subject ID":"Intra14_SER_g2_6h_mL0.05",
"Sample ID":"SER_ctrl_g2_Intra14_6h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"SER_g2","Mice Tumor SI#":"Intracranial14","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S31.d"}
},
{
"Subject ID":"Intra15_SER_g2_10h_mL0.05",
"Sample ID":"SER_ctrl_g2_Intra15_10h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"SER_g2","Mice Tumor SI#":"Intracranial15","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S32.d"}
},
{
"Subject ID":"Intra16_SER_g2_10h_mL0.05",
"Sample ID":"SER_ctrl_g2_Intra16_10h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"SER_g2","Mice Tumor SI#":"Intracranial16","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S33.d"}
},
{
"Subject ID":"Intra9_SER_g2_2h_mL0.05",
"Sample ID":"SER_ctrl_g2 _Intra9_2h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.0"},
"Additional sample data":{"Specimen Group":"SER_g2","Mice Tumor SI#":"Intracranial9","sample  quantity)":"0.05","(units)":"mL","RAW_FILE_NAME":"1108.19 GSK126_quant527_3129_Nivi_S26.d"}
},
{
"Subject ID":"SubCu1_Tum_g1_2h_mg18.135",
"Sample ID":"SubCu1_Tum_g1_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.00"},
"Additional sample data":{"Specimen Group":"Tum_g1","Mice Tumor SI#":"SubCu1_Tum","sample  quantity)":"8.135","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_045.d"}
},
{
"Subject ID":"SubCu2_Tum_g1_2h_mg16.89",
"Sample ID":"SubCu2_Tum_g1_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.00"},
"Additional sample data":{"Specimen Group":"Tum_g1","Mice Tumor SI#":"SubCu2_Tum","sample  quantity)":"16.89","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_046.d"}
},
{
"Subject ID":"SubCu3_Tum_g1_2h_mg19.13",
"Sample ID":"SubCu3_Tum_g1_2h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"2.00"},
"Additional sample data":{"Specimen Group":"Tum_g1","Mice Tumor SI#":"SubCu3_Tum","sample  quantity)":"19.13","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_047.d"}
},
{
"Subject ID":"SubCu4_Tum_g1_6h_mg20.02",
"Sample ID":"SubCu4_Tum_g1_6h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"6.00"},
"Additional sample data":{"Specimen Group":"Tum_g1","Mice Tumor SI#":"SubCu4_Tum","sample  quantity)":"20.02","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_048.d"}
},
{
"Subject ID":"SubCu5_Tum_g1_6h_mg21.3",
"Sample ID":"SubCu5_Tum_g1_6h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"6.00"},
"Additional sample data":{"Specimen Group":"Tum_g1","Mice Tumor SI#":"SubCu5_Tum","sample  quantity)":"21.3","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_049.d"}
},
{
"Subject ID":"SubCu6_Tum_g1_6h_mg18.17",
"Sample ID":"SubCu6_Tum_g1_6h",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"6.00"},
"Additional sample data":{"Specimen Group":"Tum_g1","Mice Tumor SI#":"SubCu6_Tum","sample  quantity)":"18.17","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_050.d"}
},
{
"Subject ID":"SubCu7_Tum_g1_10h_mg20.63",
"Sample ID":"SubCu7_Tum_g1_10",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"10"},
"Additional sample data":{"Specimen Group":"Tum_g1","Mice Tumor SI#":"SubCu7_Tum","sample  quantity)":"20.63","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_051.d"}
},
{
"Subject ID":"SubCu8_Tum_g1_10h_mg22",
"Sample ID":"SubCu8_Tum_g1_10",
"Factors":{"Treatment":"GSK","Gsk Exposure Time (h)":"10"},
"Additional sample data":{"Specimen Group":"Tum_g1","Mice Tumor SI#":"SubCu8_Tum","sample  quantity)":"22","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_052.d"}
},
{
"Subject ID":"SubCu9_Tum_g2_2h_mg20.28",
"Sample ID":"SubCu9_Tum_ctrl_g2_2h_",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"Tum_g2","Mice Tumor SI#":"SubCu9_Tum","sample  quantity)":"20.28","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_089.d"}
},
{
"Subject ID":"SubCu10_Tum_g2_2h_mg17.5",
"Sample ID":"SubCu10_Tum_ctrl_g2_2h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"Tum_g2","Mice Tumor SI#":"SubCu10_Tum","sample  quantity)":"17.5","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_090.d"}
},
{
"Subject ID":"SubCu11_Tum_g2_2h_mg26.15",
"Sample ID":"SubCu11_Tum_ctrl_g2_2h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"Tum_g2","Mice Tumor SI#":"SubCu11_Tum","sample  quantity)":"26.15","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_091.d"}
},
{
"Subject ID":"SubCu12_Tum_g2_6h_mg22.8",
"Sample ID":"SubCu12_Tum_ctrl_g2_6h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"Tum_g2","Mice Tumor SI#":"SubCu12_Tum","sample  quantity)":"22.8","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_092.d"}
},
{
"Subject ID":"SubCu13_Tum_g2_6h_mg23",
"Sample ID":"SubCu13_Tum_ctrl_g2_6h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"Tum_g2","Mice Tumor SI#":"SubCu13_Tum","sample  quantity)":"23","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_093.d"}
},
{
"Subject ID":"SubCu14_Tum_g2_6h_mg19.2",
"Sample ID":"SubCu14_Tum_ctrl_g2_6h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"Tum_g2","Mice Tumor SI#":"SubCu14_Tum","sample  quantity)":"19.2","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_094.d"}
},
{
"Subject ID":"SubCu15_Tum_g2_10h_mg25.13",
"Sample ID":"SubCu15_Tum_ctrl_g2_10h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"Tum_g2","Mice Tumor SI#":"SubCu15_Tum","sample  quantity)":"25.13","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_095.d"}
},
{
"Subject ID":"SubCu16_Tum_g2_10h_mg18.89",
"Sample ID":"SubCu16_Tum_ctrl_g2_10h",
"Factors":{"Treatment":"Vehicle","Gsk Exposure Time (h)":"0.00"},
"Additional sample data":{"Specimen Group":"Tum_g2","Mice Tumor SI#":"SubCu16_Tum","sample  quantity)":"18.89","(units)":"mg","RAW_FILE_NAME":"1126.19 GSK126_quant527_176_Nivi_subcu_096.d"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"All animal experiments were performed following the guidelines stipulated by the NCI-Bethesda Animal Care and Use Committee. All murine studies were performed using female albino C57BL/6 mice, 6-8 weeks of age, procured from Jackson Laboratories (Bar Harbor, ME). For sub-cutaneous tumor studies, 6x10^6 cells of stably transduced CT2A glioma cells with mCherry-firefly luciferase were injected in 100 μL PBS. For intracranial tumor studies, 1x10^2 CT2A cells with mCherry- firefly luciferase were injected in 2uL PBS. GSK126 for in vivo studies was obtained from the NCI- Drug Synthesis and Chemistry Branch and dissolved in 20% SBE-Cyclodextrin (MedChemExpress, HY-17031) pH 4-4.5 with 1N acetic acid. Vehicle was 20% SBE-Cyclodextrin pH 4-4.5 with 1N acetic acid. Water-soluble dexamethasone (Sigma Aldrich; D2915) was administered at 1mg/kg/day also by intraperitoneal injection. Anti PD-1(InVivoMAb; BE0146) or isotype control, rat IgG2a (InVivoMAb; BE0089) were also injected intraperitoneally. Subcutaneous tumor growth was measured using calipers and thereafter tumor volume was calculated using the formula for the volume of an ellipsoid given below. In the case of intracranial tumors, tumor growth was measured using the luminescence reader IVIS Ilumina and analyzed using LivingImage Software. Samples were collected from mice with three biological replicates. Serum (50 µL) was transferred to 200 μL ice-chilled (4°C) MilliQ H2O. Tissue (~16 mg) was measured from subcutaneous and intracranial samples followed by addition of 250 μL MilliQ H2O. Then, samples were sonicated at 40 amps (~30 s) until homogeneous. 80 μL of at 0.150 µg/mL debrisoquine in 60% methanol (MeOH)/40% water(aq) reagent was added. 500 μL chilled (-20°C) MeOH was added, vortexed (med) and incubated 15 min on ice. 250 μL chilled (-20°C) Chloroform was added, vortexed (high) and incubated 20 min in ice on rotating mixer. Mixture was centrifuged (13,000x g) for 18 min at 4°C. 705 μL of hydrophilic upper layer was aspirated and transferred to separate 1.5 mL microtubes, dried to completion under N2 gas sample concentrator, and stored at -80°C until LC/MS quantification of GSK126.","SAMPLE_TYPE":"Tumor cells","COLLECTION_METHOD":"Tumor Tissue/Serum Extract","COLLECTION_LOCATION":"Intracranial/Subcutaneous/Serum from Mice","COLLECTION_FREQUENCY":"Timepoints","COLLECTION_DURATION":"0, 2h, 6h and 10h","VOLUMEORAMOUNT_COLLECTED":"Serum (50 µL)/Tumor (~16 mg)","STORAGE_CONDITIONS":"-80℃","COLLECTION_VIALS":"15 mL","STORAGE_VIALS":"1.7 mL","COLLECTION_TUBE_TEMP":"On Ice","ADDITIVES":"Extraction reagent"},

"TREATMENT":{"TREATMENT_SUMMARY":"Treatment details • The mice were euthanized 2h, 6h and10h after drug treatment 27days after tumor injections and 12 days of continuous drug treatment. • The mice injected with intracranial tumors weighed approximately 16g and received 100mg/kg dose of GSK126/ vehicle injected intra-peritoneally in 100uL volume. • The mice injected with subcutaneous tumors weighed approximately 19g and received 100mg/kg dose of GSK 126/ vehicle injected intra-peritoneally in 120uL volume. • For euthanasia, the mice were injected with 200mg/kg ketamine (200uL/mouse). • Tissues harvested (frozen immediately) include: Intracranial mice a) Brain bearing tumor b) Lumbar lymph node c) Serum from cardiac bleed Subcutaneous mice a) Sub-cutaneous tumor b) Tumor draining inguinal lymph node (DLN) c) Non- tumor draining inguinal lymph node (NDLN) d) Serum from cardiac bleed Drug Preparation • Dissolved 1g of GSK 126 powder in 8.325mL DMSO to get a conc. of 120.12mg/mL, aliquot and store in -80C. • Dilute the GSK126 stock in 7 parts of Cyclodextrin (2-hydroxy-beta-cyclodextrin) to get a concentration of 15mg/mL of drug (done fresh everyday) and then inject 100uL/120uL per mouse as indicated above. • Cyclodextrin was prepared by dissolving 45g in 100mL ddH20 as indicated by manufacturer’s description. • For the vehicle control, dilute 1 part of DMSO in 7 parts of Cyclodextrin (done fresh everyday) and then inject 100uL/120uL per mouse as indicated above.","TREATMENT_PROTOCOL_ID":"Treatment","TREATMENT":"Dissolved 1g of GSK 126 powder in 8.325mL DMSO to get a conc. of 120.12mg/mL, aliquot and store in -80C. • Dilute the GSK126 stock in 7 parts of Cyclodextrin (2-hydroxy-beta-cyclodextrin) to get a concentration of 15mg/mL of drug (done fresh everyday) and then inject 100uL/120uL per mouse","TREATMENT_ROUTE":"Intra-peritoneal injection of GSK126 and/or vehicle","TREATMENT_DOSE":"100mg/kg dose of GSK 126","TREATMENT_DOSEVOLUME":"100mg/kg dose of GSK 126/120 uL","TREATMENT_DOSEDURATION":"0, 2, 6, 10h","TREATMENT_VEHICLE":"1 part of DMSO in 7 parts of Cyclodextrin (done fresh everyday) and then inject 100uL/120uL per mouse","ANIMAL_ENDP_EUTHANASIA":"Mice were injected with 200mg/kg ketamine (200uL/mouse).","ANIMAL_ENDP_TISSUE_COLL_LIST":"Intracranial mice a) Brain bearing tumor b) Lumbar lymph node c) Serum from cardiac bleed Subcutaneous mice a) Sub-cutaneous tumor b) Tumor draining inguinal lymph node (DLN) c) Non- tumor draining inguinal lymph node (NDLN) d) Serum from cardiac bleed","TREATMENT_COMPOUND":"GSK126"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Serum (50 µL) was transferred to 200 μL ice-chilled (4°C) MilliQ H2O. Tissue (~16 mg) was measured from subcutaneous and intracranial samples followed by addition of 250 μL MilliQ H2O. Then, samples were sonicated at 40 amps (~30 s) until homogeneous. 80 μL of at 0.150 µg/mL debrisoquine in 60% methanol (MeOH)/40% water(aq) reagent was added. 500 μL chilled (-20°C) MeOH was added, vortexed (med) and incubated 15 min on ice. 250 μL chilled (-20°C) Chloroform was added, vortexed (high) and incubated 20 min in ice on rotating mixer. Mixture was centrifuged (13,000x g) for 18 min at 4°C. 705 μL of hydrophilic upper layer was aspirated and transferred to separate 1.5 mL microtubes, dried to completion under N2 gas sample concentrator, and stored at -80°C","SAMPLEPREP_PROTOCOL_ID":"Extraction","PROCESSING_STORAGE_CONDITIONS":"-80℃","EXTRACTION_METHOD":"LCMS Extraction","EXTRACT_CLEANUP":"Bligh-Dyer biphasic separation, discard protein disk and dried to completion under N2 gas","SAMPLE_RESUSPENSION":"80 uL 60% MeOH (aq)","SAMPLE_SPIKING":"Internal standard 0.150 µg/mL debrisoquine(IS) was added to each"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Samples were injected at 8 µL over an 8.3 min gradient on the AdvanceBio Glycan Map 2.1 x 100 mm 2.7µm column at 35°C with a flow rate of 0.220 mL/min. The LC gradient only utilized LC/MS grade reagents when preparing mobile phases, A (88:12 H2O/acetonitrile (ACN) and B 90% ACN (aq). Both mobile phases were composed with 10 mM ammonium acetate and titrated to pH 6.85 using formic acid and ammonium hydroxide. The LC gradient was initially 100% B for 0.25 min and then ramped to 55% B at 2.5 min; 49% B at 4.5 min; 35% B at 5.5 min; 20% B at 6 min; held for 0.5 min; 15% B at 7 min; 100%B at 8.3 min followed by equilibration for 1.2 min.","CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"Agilent 1290 Infinity II","COLUMN_NAME":"Agilent AdvanceBio Glycan Map 2.1 x 100 mm 2.7µm column","FLOW_GRADIENT":"The LC gradient was initially 100% B for 0.25 min and then ramped to 55% B at 2.5 min; 49% B at 4.5 min; 35% B at 5.5 min; 20% B at 6 min; held for 0.5 min; 15% B at 7 min; 100%B at 8.3 min followed by equilibration for 1.2 min.","FLOW_RATE":"0.220 mL/min","COLUMN_TEMPERATURE":"35°C","SOLVENT_A":"88:12 H2O/acetonitrile (ACN) with 10 mM ammonium acetate and titrated to pH 6.85","SOLVENT_B":"90% ACN (aq) with 10 mM ammonium acetate and titrated to pH 6.85","COLUMN_PRESSURE":"600 bar","INJECTION_TEMPERATURE":"4°C","INTERNAL_STANDARD":"Continuous accurate mass correction was achieved by infusing proprietary Agilent Technologies API-TOF reference mass standard solution","RETENTION_TIME":"2.5-2.7 min","SAMPLE_INJECTION":"8 uL","SAMPLING_CONE":"2kV","ANALYTICAL_TIME":"8.3 min","CAPILLARY_VOLTAGE":"3 kV","SHEATH_LIQUID":"N2"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","LABORATORY_NAME":"NOB Cancer Metabolism","OPERATOR_NAME":"Dowdy","SOFTWARE_VERSION":"8.0","DATA_FORMAT":".d.zip"},

"MS":{"INSTRUMENT_NAME":"Agilent 6545","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"MS/MS Using Masshunter Qtof Quant-My-Way 10.0 software, GSK126 was detected at elution time 2.6 min using precursor ion m/z 527.3129 and transition m/z 375.2183 generated via N2 gas collision-induced fragmentation (CID) at a collision energy (CE) of 12 V. Internal standard (IS) debrisoquine detected at elution time of 2.5 mins with precursor m/z 176.1182 and transition m/z 134.0964 generated at CE 12V.","CAPILLARY_TEMPERATURE":"325°C","CAPILLARY_VOLTAGE":"3kV","COLLISION_ENERGY":"12V","COLLISION_GAS":"N2","DRY_GAS_FLOW":"9 L/min","DRY_GAS_TEMP":"250°C","FRAGMENT_VOLTAGE":"100 V","FRAGMENTATION_METHOD":"MS/MS","ION_SOURCE_TEMPERATURE":"325°C","MASS_ACCURACY":"2 kDa","SOURCE_TEMPERATURE":"325°C","CDL_TEMPERATURE":"RT","DESOLVATION_TEMPERATURE":"325°C","NEBULIZER":"45 psig","OCTPOLE_VOLTAGE":"750V"},

"MS_METABOLITE_DATA":{
"Units":"ng",

"Data":[{"Metabolite":"GSK126","Blank_S14.d":"0","blank_S22.d":"0","blank_Sr62.d":"0","BRN_g1 _Intra1_2h":"0","BRN_g1 _Intra2_2h":"41.768","BRN_g1 _Intra3_2h":"25.344","BRN_g1 _Intra4_6h":"0","BRN_g1 _Intra5_6h":"13.712","BRN_g1 _Intra7_10h":"13.744","BRN_g1 _Intra8_10h":"0","BRN_ctrl_g2_Intra10_2h":"0","BRN_ctrl_g2_Intra11_2h":"0","BRN_ctrl_g2_Intra12_6h":"0","BRN_ctrl_g2_Intra13_6h":"0","BRN_ctrl_g2_Intra14_6h":"0","BRN_ctrl_g2_Intra15_10h":"0","BRN_ctrl_g2_Intra16_10h":"0","SubCu1_DLN_g1_2h":"126.35","SubCu2_DLN_g1_2h":"449.75","SubCu3_DLN_g1_2h":"677.39","SubCu4_DLN_g1_6h":"2245.46","SubCu5_DLN_g1_6h":"765.45","SubCu6_DLN_g1_6h":"530.67","SubCu7_DLN_g1_10":"502.53","SubCu8_DLN_g1_10":"746.97","SubCu9_DLN_ctrl_g2_2h_":"0","SubCu10_DLN_ctrl_g2_2h":"0","SubCu11_DLN_ctrl_g2_2h":"0","SubCu12_DLN_ctrl_g2_6h":"0","SubCu13_DLN_ctrl_g2_6h":"0","SubCu15_DLN_ctrl_g2_10h":"0","SubCu16_DLN_ctrl_g2_10h":"0","LLN_g1 _Intra1_2h":"311.784","LLN_g1 _Intra2_2h":"605.704","LLN_g1 _Intra3_2h":"156.192","LLN_g1 _Intra4_6h":"31.888","LLN_g1 _Intra6_6h":"20.512","LLN_g1 _Intra7_10h":"22.808","LLN_g1 _Intra8_10h":"42.184","LLN_ctrl_g2_Intra10_2h":"0","LLN_ctrl_g2_Intra11_2h":"0","LLN_ctrl_g2_Intra12_6h":"0","LLN_ctrl_g2_Intra13_6h":"0","LLN_ctrl_g2_Intra15_10h":"0","LLN_ctrl_g2_Intra16_10h":"0","LLN_ctrl_g2 _Intra9_2h":"0","SubCu1_NLN_g1_2h":"189.42","SubCu2_NLN_g1_2h":"255.78","SubCu3_NLN_g1_2h":"295.4","SubCu4_NLN_g1_6h":"1089.55","SubCu5_NLN_g1_6h":"415.1","SubCu6_NLN_g1_6h":"553.98","SubCu7_NLN_g1_10":"1230.67","SubCu8_NLN_g1_10":"459.97","SubCu9_NLN_ctrl_g2_2h_":"0","SubCu10_NLN_ctrl_g2_2h":"0","SubCu11_NLN_ctrl_g2_2h":"0","SubCu12_NLN_ctrl_g2_6h":"0","SubCu13_NLN_ctrl_g2_6h":"0","SubCu14_NLN_ctrl_g2_6h":"0","SubCu15_NLN_ctrl_g2_10h":"0","SubCu16_NLN_ctrl_g2_10h":"0","QcBRN_S45.d":"0","QcBRN_Sr91.d":"0","QcBRN_Sr111.d":"0","QcBRN_Sr65.d":"0","QcLLN_L1002.d":"69.28","QCpool g1 D_NLN_subcu_055.d":"634.83","QCpool g1 D_NLN_subcu_081.d":"637.42","QCpool g1 D_NLN_subcu_135.d":"640.01","QCpool g1 D_NLN_subcu_012.d":"605.64","QCpool g1 D_NLN_subcu_028.d":"597.24","QCpool g1 D_NLN_subcu_037.d":"615.65","QCpool g1 SubTum_038.d":"361.27","QCpool g1 SubTum_056.d":"453.39","QCpool g1 SubTum_082.d":"354.97","QCpool g1 SubTum_136.d":"416.29","QcSer_S24.d":"41.216","QcSer_S44.d":"45.976","SER_g1 _Subcutaneous1_2h":"72.528","SER_g1 _Subcutaneous2_2h":"22.848","SER_g1 _Subcutaneous3_2h":"91.624","SER_g1 _Subcutaneous4_6h":"24.312","SER_g1 _Subcutaneous5_6h":"46.088","SER_g1 _Subcutaneous6_6h":"55.04","SER_g1 _Subcutaneous7_10h":"0","SER_g1 _Subcutaneous8_10h":"32","SER_g1 _Intra1_2h":"160.768","SER_g1 _Intra2_2h":"220.832","SER_g1 _Intra3_2h":"149.024","SER_g1 _Intra4_6h":"0","SER_g1 _Intra7_10h":"41.792","SER_g1 _Intra8_10h":"14.352","SER_g1 _Intra5_6h":"147.568","SER_g1 _Intra6_6h":"15.256","SER_ctrl_g2_Subcutaneous10_2h":"0","SER_ctrl_g2_Subcutaneous12_6h":"0","SER_ctrl_g2_Subcutaneous13_6h":"0","SER_ctrl_g2_Subcutaneous14_6h":"0","SER_ctrl_g2_Subcutaneous15_10h":"0","SER_ctrl_g2_Subcutaneous16_10h":"0","SER_ctrl_g2_Intra10_2h":"0","SER_ctrl_g2_Intra11_2h":"0","SER_ctrl_g2_Intra12_6h":"0","SER_ctrl_g2_Intra13_6h":"0","SER_ctrl_g2_Intra14_6h":"0","SER_ctrl_g2_Intra15_10h":"0","SER_ctrl_g2_Intra16_10h":"0","SER_ctrl_g2 _Intra9_2h":"0","SubCu1_Tum_g1_2h":"694.4","SubCu2_Tum_g1_2h":"0","SubCu3_Tum_g1_2h":"248.36","SubCu4_Tum_g1_6h":"267.19","SubCu5_Tum_g1_6h":"225.05","SubCu6_Tum_g1_6h":"1329.09","SubCu7_Tum_g1_10":"107.24","SubCu8_Tum_g1_10":"223.72","SubCu9_Tum_ctrl_g2_2h_":"0","SubCu10_Tum_ctrl_g2_2h":"0","SubCu11_Tum_ctrl_g2_2h":"0","SubCu12_Tum_ctrl_g2_6h":"0","SubCu13_Tum_ctrl_g2_6h":"0","SubCu14_Tum_ctrl_g2_6h":"0","SubCu15_Tum_ctrl_g2_10h":"0","SubCu16_Tum_ctrl_g2_10h":"0"}],

"Metabolites":[{"Metabolite":"GSK126","retention index":"","quantified m/z":"527.3129","PubChem ID":"68210102","KEGG ID":"K11430"}]
}

}