#METABOLOMICS WORKBENCH dowdytm_20210611_111543 DATATRACK_ID:2684 STUDY_ID:ST001838 ANALYSIS_ID:AN002980 PROJECT_ID:PR001160
VERSION             	1
CREATED_ON             	June 24, 2021, 2:45 pm
#PROJECT
PR:PROJECT_TITLE                 	Reversing Epigenetic Gene Silencing to Overcome Immune Evasion in CNS
PR:PROJECT_TITLE                 	Malignancies
PR:PROJECT_SUMMARY               	Glioblastoma is an aggressive brain malignancy with a dismal prognosis. With
PR:PROJECT_SUMMARY               	emerging evidence that disproves the immune privileged environment in the brain,
PR:PROJECT_SUMMARY               	there is much interest in examining various immunotherapy strategies to treat
PR:PROJECT_SUMMARY               	these incurable cancers. Unfortunately, to date, clinical studies investigating
PR:PROJECT_SUMMARY               	immunotherapy regimens have not provided much evidence of efficacy, leading to
PR:PROJECT_SUMMARY               	questions about the suitability of immunotherapy strategies for these tumors.
PR:PROJECT_SUMMARY               	Inadequate inherent populations of lymphocytes in tumor (TILs) and limited
PR:PROJECT_SUMMARY               	trafficking of systemic circulating T cells into the central nervous system
PR:PROJECT_SUMMARY               	(CNS) likely contribute to the poor response to immunotherapy treatment for
PR:PROJECT_SUMMARY               	primary CNS cancers. This paucity of TILs is in concert with the finding of
PR:PROJECT_SUMMARY               	epigenetic silencing of genes that promote immune cell movement (chemotaxis) to
PR:PROJECT_SUMMARY               	the tumor. In this study we evaluated the ability of GSK126, a blood-brain
PR:PROJECT_SUMMARY               	barrier permeable small molecule inhibitor of EZH2, to reverse the epigenetic
PR:PROJECT_SUMMARY               	silencing of chemokines like CXCL9 and CXCL10. When combined with anti-PD-1
PR:PROJECT_SUMMARY               	treatment, these IFN driven chemokines promote T cell infiltration, resulting in
PR:PROJECT_SUMMARY               	decreased tumor growth and enhanced survival in immunocompetent murine
PR:PROJECT_SUMMARY               	sub-cutaneous and intracranial tumor syngeneic models of GBM. Examination of the
PR:PROJECT_SUMMARY               	tumor micro-environment revealed that the decrease in tumor growth in the mice
PR:PROJECT_SUMMARY               	treated with the drug combination was accompanied by increased tumor CD8 T cell
PR:PROJECT_SUMMARY               	infiltration along with higher IFN expression. Additionally, a significant
PR:PROJECT_SUMMARY               	increase in CXCR3+ T cells in the draining lymph nodes was also found. Taken
PR:PROJECT_SUMMARY               	together, our data suggests that in glioblastoma, epigenetic modulation using
PR:PROJECT_SUMMARY               	GSK126 could improve current immunotherapy strategies by reversing the
PR:PROJECT_SUMMARY               	epigenetic changes that enable immune cell evasion leading to enhanced immune
PR:PROJECT_SUMMARY               	c-ll trafficking to the tumor.
PR:INSTITUTE                     	National Cancer Institute
PR:DEPARTMENT                    	Neuro-Oncology Branch
PR:LABORATORY                    	Cancer Metabolism
PR:LAST_NAME                     	Dowdy
PR:FIRST_NAME                    	Tyrone
PR:ADDRESS                       	37 convent dr, Bldg 37 rm 1142
PR:EMAIL                         	tyrone.dowdy@nih.gov
PR:PHONE                         	2407607066
PR:CONTRIBUTORS                  	Nivedita M. Ratnam1, Heather M. Sonnemann1, Stephen C. Frederico1, Huanwen
PR:CONTRIBUTORS                  	Chen1, Marsha-Kay N.D. Hutchinson1, Tyrone Dowdy1, Caitlin M. Reid1, Jinkyu
PR:CONTRIBUTORS                  	Jung1, Wei Zhang1, Hua Song1, Meili Zhang1, Dionne Davis1, Mioara Larion1, Amber
PR:CONTRIBUTORS                  	J. Giles1 and Mark R. Gilbert1*.
#STUDY
ST:STUDY_TITLE                   	Reversing Epigenetic Gene Silencing to Overcome Immune Evasion in CNS
ST:STUDY_TITLE                   	Malignancies
ST:STUDY_SUMMARY                 	Glioblastoma is an aggressive brain malignancy with a dismal prognosis. With
ST:STUDY_SUMMARY                 	emerging evidence that disproves the immune privileged environment in the brain,
ST:STUDY_SUMMARY                 	there is much interest in examining various immunotherapy strategies to treat
ST:STUDY_SUMMARY                 	these incurable cancers. Unfortunately, to date, clinical studies investigating
ST:STUDY_SUMMARY                 	immunotherapy regimens have not provided much evidence of efficacy, leading to
ST:STUDY_SUMMARY                 	questions about the suitability of immunotherapy strategies for these tumors.
ST:STUDY_SUMMARY                 	Inadequate inherent populations of lymphocytes in tumor (TILs) and limited
ST:STUDY_SUMMARY                 	trafficking of systemic circulating T cells into the central nervous system
ST:STUDY_SUMMARY                 	(CNS) likely contribute to the poor response to immunotherapy treatment for
ST:STUDY_SUMMARY                 	primary CNS cancers. This paucity of TILs is in concert with the finding of
ST:STUDY_SUMMARY                 	epigenetic silencing of genes that promote immune cell movement (chemotaxis) to
ST:STUDY_SUMMARY                 	the tumor. In this study we evaluated the ability of GSK126, a blood-brain
ST:STUDY_SUMMARY                 	barrier permeable small molecule inhibitor of EZH2, to reverse the epigenetic
ST:STUDY_SUMMARY                 	silencing of chemokines like CXCL9 and CXCL10. When combined with anti-PD-1
ST:STUDY_SUMMARY                 	treatment, these IFN driven chemokines promote T cell infiltration, resulting in
ST:STUDY_SUMMARY                 	decreased tumor growth and enhanced survival in immunocompetent murine
ST:STUDY_SUMMARY                 	sub-cutaneous and intracranial tumor syngeneic models of GBM. Examination of the
ST:STUDY_SUMMARY                 	tumor micro-environment revealed that the decrease in tumor growth in the mice
ST:STUDY_SUMMARY                 	treated with the drug combination was accompanied by increased tumor CD8 T cell
ST:STUDY_SUMMARY                 	infiltration along with higher IFN expression. Additionally, a significant
ST:STUDY_SUMMARY                 	increase in CXCR3+ T cells in the draining lymph nodes was also found. Taken
ST:STUDY_SUMMARY                 	together, our data suggests that in glioblastoma, epigenetic modulation using
ST:STUDY_SUMMARY                 	GSK126 could improve current immunotherapy strategies by reversing the
ST:STUDY_SUMMARY                 	epigenetic changes that enable immune cell evasion leading to enhanced immune
ST:STUDY_SUMMARY                 	cell trafficking to the tumor.
ST:INSTITUTE                     	National Cancer Institute
ST:DEPARTMENT                    	Neuro-Oncology Branch
ST:LABORATORY                    	Cancer Metabolism
ST:LAST_NAME                     	Dowdy
ST:FIRST_NAME                    	Tyrone
ST:ADDRESS                       	37 convent dr, Bldg 37 rm 1142
ST:EMAIL                         	tyrone.dowdy@nih.gov
ST:PHONE                         	2407607066
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
SU:GENOTYPE_STRAIN               	albino C57BL/6
SU:AGE_OR_AGE_RANGE              	6-8 weeks
SU:GENDER                        	Female
SU:ANIMAL_HOUSING                	NCI-Bethesda Animal Facility
SU:ANIMAL_ANIMAL_SUPPLIER        	Jackson Laboratories (Bar Harbor, ME)
#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           	blank	Blank_S14.d	Treatment:blank | Gsk Exposure Time (h):-	Specimen Group=blank; Mice Tumor SI#=blank; sample  quantity)=-; (units)=blank; RAW_FILE_NAME=1108.19 GSK126_quant527_3129_Nivi_S14.d
SUBJECT_SAMPLE_FACTORS           	blank	blank_S22.d	Treatment:blank | Gsk Exposure Time (h):-	Specimen Group=blank; Mice Tumor SI#=blank; sample  quantity)=-; (units)=blank; RAW_FILE_NAME=1108.19 GSK126_quant527_3129_Nivi_S22.d
SUBJECT_SAMPLE_FACTORS           	blank	blank_Sr62.d	Treatment:blank | Gsk Exposure Time (h):-	Specimen Group=blank; Mice Tumor SI#=blank; sample  quantity)=-; (units)=blank; RAW_FILE_NAME=1108.19 GSK126_quant527_3129_Nivi_Sr62blk.d
SUBJECT_SAMPLE_FACTORS           	Intra1_BRN_g1_2h_mg19.335	BRN_g1 _Intra1_2h	Treatment:GSK | Gsk Exposure Time (h):2.0	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_SAMPLE_FACTORS           	Intra2_BRN_g1_2h_mg19.835	BRN_g1 _Intra2_2h	Treatment:GSK | Gsk Exposure Time (h):2.0	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_SAMPLE_FACTORS           	Intra3_BRN_g1_2h_mg20.96	BRN_g1 _Intra3_2h	Treatment:GSK | Gsk Exposure Time (h):2.0	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_SAMPLE_FACTORS           	Intra4_BRN_g1_6h_mg25.175	BRN_g1 _Intra4_6h	Treatment:GSK | Gsk Exposure Time (h):6.0	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_SAMPLE_FACTORS           	Intra5_BRN_g1_6h_mg16.81	BRN_g1 _Intra5_6h	Treatment:GSK | Gsk Exposure Time (h):6.0	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_SAMPLE_FACTORS           	Intra7_BRN_g1_10h_mg18.28	BRN_g1 _Intra7_10h	Treatment:GSK | Gsk Exposure Time (h):10.0	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_SAMPLE_FACTORS           	Intra8_BRN_g1_10h_mg16	BRN_g1 _Intra8_10h	Treatment:GSK | Gsk Exposure Time (h):10.0	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_SAMPLE_FACTORS           	Intra10_BRN_g2_2h_mg22.06	BRN_ctrl_g2_Intra10_2h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra11_BRN_g2_2h_mg16.44	BRN_ctrl_g2_Intra11_2h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra12_BRN_g2_6h_mg18.26	BRN_ctrl_g2_Intra12_6h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra13_BRN_g2_6h_mg18.35	BRN_ctrl_g2_Intra13_6h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra14_BRN_g2_6h_mg16	BRN_ctrl_g2_Intra14_6h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra15_BRN_g2_10h_mg21.2	BRN_ctrl_g2_Intra15_10h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra16_BRN_g2_10h_mg18.6	BRN_ctrl_g2_Intra16_10h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	SubCu1_DLN_g1_2h_mg1.92	SubCu1_DLN_g1_2h	Treatment:GSK | Gsk Exposure Time (h):2.00	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_SAMPLE_FACTORS           	SubCu2_DLN_g1_2h_mg8.04	SubCu2_DLN_g1_2h	Treatment:GSK | Gsk Exposure Time (h):2.00	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_SAMPLE_FACTORS           	SubCu3_DLN_g1_2h_mg2.945	SubCu3_DLN_g1_2h	Treatment:GSK | Gsk Exposure Time (h):2.00	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_SAMPLE_FACTORS           	SubCu4_DLN_g1_6h_mg1.82	SubCu4_DLN_g1_6h	Treatment:GSK | Gsk Exposure Time (h):6.00	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_SAMPLE_FACTORS           	SubCu5_DLN_g1_6h_mg3.145	SubCu5_DLN_g1_6h	Treatment:GSK | Gsk Exposure Time (h):6.00	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_SAMPLE_FACTORS           	SubCu6_DLN_g1_6h_mg2.21	SubCu6_DLN_g1_6h	Treatment:GSK | Gsk Exposure Time (h):6.00	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_SAMPLE_FACTORS           	SubCu7_DLN_g1_10h_mg1.5	SubCu7_DLN_g1_10	Treatment:GSK | Gsk Exposure Time (h):10	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_SAMPLE_FACTORS           	SubCu8_DLN_g1_10h_mg3	SubCu8_DLN_g1_10	Treatment:GSK | Gsk Exposure Time (h):10	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_SAMPLE_FACTORS           	SubCu9_DLN_g2_2h_mg10.475	SubCu9_DLN_ctrl_g2_2h_	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu10_DLN_g2_2h_mg3.07	SubCu10_DLN_ctrl_g2_2h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu11_DLN_g2_2h_mg2.49	SubCu11_DLN_ctrl_g2_2h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu12_DLN_g2_6h_mg4.505	SubCu12_DLN_ctrl_g2_6h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu13_DLN_g2_6h_mg12.535	SubCu13_DLN_ctrl_g2_6h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu15_DLN_g2_10h_mg6.26	SubCu15_DLN_ctrl_g2_10h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu16_DLN_g2_10h_mg5.39	SubCu16_DLN_ctrl_g2_10h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	Intra1_LLN_g1_2h_mg3.05	LLN_g1 _Intra1_2h	Treatment:GSK | Gsk Exposure Time (h):2.0	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_SAMPLE_FACTORS           	Intra2_LLN_g1_2h_mg5	LLN_g1 _Intra2_2h	Treatment:GSK | Gsk Exposure Time (h):2.0	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_SAMPLE_FACTORS           	Intra3_LLN_g1_2h_mg1.975	LLN_g1 _Intra3_2h	Treatment:GSK | Gsk Exposure Time (h):2.0	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_SAMPLE_FACTORS           	Intra4_LLN_g1_6h_mg1.03	LLN_g1 _Intra4_6h	Treatment:GSK | Gsk Exposure Time (h):6.0	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_SAMPLE_FACTORS           	Intra6_LLN_g1_6h_mg0.975	LLN_g1 _Intra6_6h	Treatment:GSK | Gsk Exposure Time (h):6.0	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_SAMPLE_FACTORS           	Intra7_LLN_g1_10h_mg0.975	LLN_g1 _Intra7_10h	Treatment:GSK | Gsk Exposure Time (h):10.0	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_SAMPLE_FACTORS           	Intra8_LLN_g1_10h_mg1.935	LLN_g1 _Intra8_10h	Treatment:GSK | Gsk Exposure Time (h):10.0	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_SAMPLE_FACTORS           	Intra10_LLN_g2_2h_mg0.87	LLN_ctrl_g2_Intra10_2h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra11_LLN_g2_2h_mg1.65	LLN_ctrl_g2_Intra11_2h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra12_LLN_g2_6h_mg0.965	LLN_ctrl_g2_Intra12_6h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra13_LLN_g2_6h_mg2.885	LLN_ctrl_g2_Intra13_6h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra15_LLN_g2_10h_mg1.345	LLN_ctrl_g2_Intra15_10h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra16_LLN_g2_10h_mg1.25	LLN_ctrl_g2_Intra16_10h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra9_LLN_g2_2h_mg0.8	LLN_ctrl_g2 _Intra9_2h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	SubCu1_NLN_g1_2h_mg2.08	SubCu1_NLN_g1_2h	Treatment:GSK | Gsk Exposure Time (h):2.00	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_SAMPLE_FACTORS           	SubCu2_NLN_g1_2h_mg5.82	SubCu2_NLN_g1_2h	Treatment:GSK | Gsk Exposure Time (h):2.00	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_SAMPLE_FACTORS           	SubCu3_NLN_g1_2h_mg2.03	SubCu3_NLN_g1_2h	Treatment:GSK | Gsk Exposure Time (h):2.00	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_SAMPLE_FACTORS           	SubCu4_NLN_g1_6h_mg3.64	SubCu4_NLN_g1_6h	Treatment:GSK | Gsk Exposure Time (h):6.00	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_SAMPLE_FACTORS           	SubCu5_NLN_g1_6h_mg1.8	SubCu5_NLN_g1_6h	Treatment:GSK | Gsk Exposure Time (h):6.00	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_SAMPLE_FACTORS           	SubCu6_NLN_g1_6h_mg1.62	SubCu6_NLN_g1_6h	Treatment:GSK | Gsk Exposure Time (h):6.00	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_SAMPLE_FACTORS           	SubCu7_NLN_g1_10h_mg2.36	SubCu7_NLN_g1_10	Treatment:GSK | Gsk Exposure Time (h):10	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_SAMPLE_FACTORS           	SubCu8_NLN_g1_10h_mg3.48	SubCu8_NLN_g1_10	Treatment:GSK | Gsk Exposure Time (h):10	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_SAMPLE_FACTORS           	SubCu9_NLN_g2_2h_mg4.45	SubCu9_NLN_ctrl_g2_2h_	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu10_NLN_g2_2h_mg2.58	SubCu10_NLN_ctrl_g2_2h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu11_NLN_g2_2h_mg4.5	SubCu11_NLN_ctrl_g2_2h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu12_NLN_g2_6h_mg7.29	SubCu12_NLN_ctrl_g2_6h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu13_NLN_g2_6h_mg3.37	SubCu13_NLN_ctrl_g2_6h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu14_NLN_g2_6h_mg8.03	SubCu14_NLN_ctrl_g2_6h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu15_NLN_g2_10h_mg2.81	SubCu15_NLN_ctrl_g2_10h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu16_NLN_g2_10h_mg3.16	SubCu16_NLN_ctrl_g2_10h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	QcBRN	QcBRN_S45.d	Treatment:QC | Gsk Exposure Time (h):-	Specimen Group=QcBRN; Mice Tumor SI#=QcBRN; sample  quantity)=-; (units)=Qc; RAW_FILE_NAME=1108.19 GSK126_quant527_3129_Nivi_S45.d
SUBJECT_SAMPLE_FACTORS           	QcBRN	QcBRN_Sr91.d	Treatment:QC | Gsk Exposure Time (h):-	Specimen Group=QcBRN; Mice Tumor SI#=QcBRN; sample  quantity)=-; (units)=Qc; RAW_FILE_NAME=1108.19 GSK126_quant527_3129_Nivi_Sr91.d
SUBJECT_SAMPLE_FACTORS           	QcBRN	QcBRN_Sr111.d	Treatment:QC | Gsk Exposure Time (h):-	Specimen Group=QcBRN; Mice Tumor SI#=QcBRN; sample  quantity)=-; (units)=Qc; RAW_FILE_NAME=1108.19 GSK126_quant527_3129_Nivi_Sr111.d
SUBJECT_SAMPLE_FACTORS           	QcBRN	QcBRN_Sr65.d	Treatment:QC | Gsk Exposure Time (h):-	Specimen Group=QcBRN; Mice Tumor SI#=QcBRN; sample  quantity)=-; (units)=Qc; RAW_FILE_NAME=1108.19 GSK126_quant527_3129_Nivi_Sr65.d
SUBJECT_SAMPLE_FACTORS           	QcLLN	QcLLN_L1002.d	Treatment:QC | Gsk Exposure Time (h):-	Specimen Group=QcLLN_Intra; Mice Tumor SI#=QcLLN; sample  quantity)=-; (units)=Qc; RAW_FILE_NAME=1108.19 GSK126_quant527_3129_Nivi_L1002.d
SUBJECT_SAMPLE_FACTORS           	QCpool g1 D_NLN	QCpool g1 D_NLN_subcu_055.d	Treatment:QC | Gsk Exposure Time (h):-	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_SAMPLE_FACTORS           	QCpool g1 D_NLN	QCpool g1 D_NLN_subcu_081.d	Treatment:QC | Gsk Exposure Time (h):-	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_SAMPLE_FACTORS           	QCpool g1 D_NLN	QCpool g1 D_NLN_subcu_135.d	Treatment:QC | Gsk Exposure Time (h):-	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_SAMPLE_FACTORS           	QCpool g1 D_NLN	QCpool g1 D_NLN_subcu_012.d	Treatment:QC | Gsk Exposure Time (h):-	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_SAMPLE_FACTORS           	QCpool g1 D_NLN	QCpool g1 D_NLN_subcu_028.d	Treatment:QC | Gsk Exposure Time (h):-	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_SAMPLE_FACTORS           	QCpool g1 D_NLN	QCpool g1 D_NLN_subcu_037.d	Treatment:QC | Gsk Exposure Time (h):-	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_SAMPLE_FACTORS           	QCpool g1 SubTum	QCpool g1 SubTum_038.d	Treatment:QC | Gsk Exposure Time (h):-	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_SAMPLE_FACTORS           	QCpool g1 SubTum	QCpool g1 SubTum_056.d	Treatment:QC | Gsk Exposure Time (h):-	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_SAMPLE_FACTORS           	QCpool g1 SubTum	QCpool g1 SubTum_082.d	Treatment:QC | Gsk Exposure Time (h):-	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_SAMPLE_FACTORS           	QCpool g1 SubTum	QCpool g1 SubTum_136.d	Treatment:QC | Gsk Exposure Time (h):-	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_SAMPLE_FACTORS           	QcSer	QcSer_S24.d	Treatment:QC | Gsk Exposure Time (h):-	Specimen Group=QcSer; Mice Tumor SI#=QcSer_; sample  quantity)=-; (units)=Qc; RAW_FILE_NAME=1108.19 GSK126_quant527_3129_Nivi_S24.d
SUBJECT_SAMPLE_FACTORS           	QcSer	QcSer_S44.d	Treatment:QC | Gsk Exposure Time (h):-	Specimen Group=QcSer; Mice Tumor SI#=QcSer; sample  quantity)=-; (units)=Qc; RAW_FILE_NAME=1108.19 GSK126_quant527_3129_Nivi_S44.d
SUBJECT_SAMPLE_FACTORS           	Subcutaneous1_SER_g1_2h_mL0.05	SER_g1 _Subcutaneous1_2h	Treatment:GSK | Gsk Exposure Time (h):2.0	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_SAMPLE_FACTORS           	Subcutaneous2_SER_g1_2h_mL0.05	SER_g1 _Subcutaneous2_2h	Treatment:GSK | Gsk Exposure Time (h):2.0	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_SAMPLE_FACTORS           	Subcutaneous3_SER_g1_2h_mL0.05	SER_g1 _Subcutaneous3_2h	Treatment:GSK | Gsk Exposure Time (h):2.0	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_SAMPLE_FACTORS           	Subcutaneous4_SER_g1_6h_mL0.05	SER_g1 _Subcutaneous4_6h	Treatment:GSK | Gsk Exposure Time (h):6.0	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_SAMPLE_FACTORS           	Subcutaneous5_SER_g1_6h_mL0.05	SER_g1 _Subcutaneous5_6h	Treatment:GSK | Gsk Exposure Time (h):6.0	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_SAMPLE_FACTORS           	Subcutaneous6_SER_g1_6h_mL0.05	SER_g1 _Subcutaneous6_6h	Treatment:GSK | Gsk Exposure Time (h):6.0	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_SAMPLE_FACTORS           	Subcutaneous7_SER_g1_10h_mL0.05	SER_g1 _Subcutaneous7_10h	Treatment:GSK | Gsk Exposure Time (h):10.0	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_SAMPLE_FACTORS           	Subcutaneous8_SER_g1_10h_mL0.05	SER_g1 _Subcutaneous8_10h	Treatment:GSK | Gsk Exposure Time (h):10.0	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_SAMPLE_FACTORS           	Intra1_SER_g1_2h_mL0.05	SER_g1 _Intra1_2h	Treatment:GSK | Gsk Exposure Time (h):2.0	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_SAMPLE_FACTORS           	Intra2_SER_g1_2h_mL0.05	SER_g1 _Intra2_2h	Treatment:GSK | Gsk Exposure Time (h):2.0	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_SAMPLE_FACTORS           	Intra3_SER_g1_2h_mL0.05	SER_g1 _Intra3_2h	Treatment:GSK | Gsk Exposure Time (h):2.0	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_SAMPLE_FACTORS           	Intra4_SER_g1_6h_mL0.05	SER_g1 _Intra4_6h	Treatment:GSK | Gsk Exposure Time (h):6.0	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_SAMPLE_FACTORS           	Intra7_SER_g1_10h_mL0.05	SER_g1 _Intra7_10h	Treatment:GSK | Gsk Exposure Time (h):10.0	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_SAMPLE_FACTORS           	Intra8_SER_g1_10h_mL0.05	SER_g1 _Intra8_10h	Treatment:GSK | Gsk Exposure Time (h):10.0	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_SAMPLE_FACTORS           	Intra5_SER_g1_6h_mL0.05	SER_g1 _Intra5_6h	Treatment:GSK | Gsk Exposure Time (h):6.0	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_SAMPLE_FACTORS           	Intra6_SER_g1_6h_mL0.05	SER_g1 _Intra6_6h	Treatment:GSK | Gsk Exposure Time (h):6.0	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_SAMPLE_FACTORS           	Subcutaneous10_SER_g2_2h_mL0.05	SER_ctrl_g2_Subcutaneous10_2h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Subcutaneous12_SER_g2_6h_mL0.05	SER_ctrl_g2_Subcutaneous12_6h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Subcutaneous13_SER_g2_6h_mL0.05	SER_ctrl_g2_Subcutaneous13_6h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Subcutaneous14_SER_g2_6h_mL0.05	SER_ctrl_g2_Subcutaneous14_6h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Subcutaneous15_SER_g2_10h_mL0.05	SER_ctrl_g2_Subcutaneous15_10h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Subcutaneous16_SER_g2_10h_mL0.05	SER_ctrl_g2_Subcutaneous16_10h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra10_SER_g2_2h_mL0.05	SER_ctrl_g2_Intra10_2h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra11_SER_g2_2h_mL0.05	SER_ctrl_g2_Intra11_2h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra12_SER_g2_6h_mL0.05	SER_ctrl_g2_Intra12_6h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra13_SER_g2_6h_mL0.05	SER_ctrl_g2_Intra13_6h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra14_SER_g2_6h_mL0.05	SER_ctrl_g2_Intra14_6h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra15_SER_g2_10h_mL0.05	SER_ctrl_g2_Intra15_10h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra16_SER_g2_10h_mL0.05	SER_ctrl_g2_Intra16_10h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	Intra9_SER_g2_2h_mL0.05	SER_ctrl_g2 _Intra9_2h	Treatment:Vehicle | Gsk Exposure Time (h):0.0	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_SAMPLE_FACTORS           	SubCu1_Tum_g1_2h_mg18.135	SubCu1_Tum_g1_2h	Treatment:GSK | Gsk Exposure Time (h):2.00	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_SAMPLE_FACTORS           	SubCu2_Tum_g1_2h_mg16.89	SubCu2_Tum_g1_2h	Treatment:GSK | Gsk Exposure Time (h):2.00	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_SAMPLE_FACTORS           	SubCu3_Tum_g1_2h_mg19.13	SubCu3_Tum_g1_2h	Treatment:GSK | Gsk Exposure Time (h):2.00	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_SAMPLE_FACTORS           	SubCu4_Tum_g1_6h_mg20.02	SubCu4_Tum_g1_6h	Treatment:GSK | Gsk Exposure Time (h):6.00	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_SAMPLE_FACTORS           	SubCu5_Tum_g1_6h_mg21.3	SubCu5_Tum_g1_6h	Treatment:GSK | Gsk Exposure Time (h):6.00	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_SAMPLE_FACTORS           	SubCu6_Tum_g1_6h_mg18.17	SubCu6_Tum_g1_6h	Treatment:GSK | Gsk Exposure Time (h):6.00	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_SAMPLE_FACTORS           	SubCu7_Tum_g1_10h_mg20.63	SubCu7_Tum_g1_10	Treatment:GSK | Gsk Exposure Time (h):10	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_SAMPLE_FACTORS           	SubCu8_Tum_g1_10h_mg22	SubCu8_Tum_g1_10	Treatment:GSK | Gsk Exposure Time (h):10	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_SAMPLE_FACTORS           	SubCu9_Tum_g2_2h_mg20.28	SubCu9_Tum_ctrl_g2_2h_	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu10_Tum_g2_2h_mg17.5	SubCu10_Tum_ctrl_g2_2h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu11_Tum_g2_2h_mg26.15	SubCu11_Tum_ctrl_g2_2h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu12_Tum_g2_6h_mg22.8	SubCu12_Tum_ctrl_g2_6h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu13_Tum_g2_6h_mg23	SubCu13_Tum_ctrl_g2_6h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu14_Tum_g2_6h_mg19.2	SubCu14_Tum_ctrl_g2_6h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu15_Tum_g2_10h_mg25.13	SubCu15_Tum_ctrl_g2_10h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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_SAMPLE_FACTORS           	SubCu16_Tum_g2_10h_mg18.89	SubCu16_Tum_ctrl_g2_10h	Treatment:Vehicle | Gsk Exposure Time (h):0.00	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
CO:COLLECTION_SUMMARY            	All animal experiments were performed following the guidelines stipulated by the
CO:COLLECTION_SUMMARY            	NCI-Bethesda Animal Care and Use Committee. All murine studies were performed
CO:COLLECTION_SUMMARY            	using female albino C57BL/6 mice, 6-8 weeks of age, procured from Jackson
CO:COLLECTION_SUMMARY            	Laboratories (Bar Harbor, ME). For sub-cutaneous tumor studies, 6x10^6 cells of
CO:COLLECTION_SUMMARY            	stably transduced CT2A glioma cells with mCherry-firefly luciferase were
CO:COLLECTION_SUMMARY            	injected in 100 μL PBS. For intracranial tumor studies, 1x10^2 CT2A cells with
CO:COLLECTION_SUMMARY            	mCherry- firefly luciferase were injected in 2uL PBS. GSK126 for in vivo studies
CO:COLLECTION_SUMMARY            	was obtained from the NCI- Drug Synthesis and Chemistry Branch and dissolved in
CO:COLLECTION_SUMMARY            	20% SBE-Cyclodextrin (MedChemExpress, HY-17031) pH 4-4.5 with 1N acetic acid.
CO:COLLECTION_SUMMARY            	Vehicle was 20% SBE-Cyclodextrin pH 4-4.5 with 1N acetic acid. Water-soluble
CO:COLLECTION_SUMMARY            	dexamethasone (Sigma Aldrich; D2915) was administered at 1mg/kg/day also by
CO:COLLECTION_SUMMARY            	intraperitoneal injection. Anti PD-1(InVivoMAb; BE0146) or isotype control, rat
CO:COLLECTION_SUMMARY            	IgG2a (InVivoMAb; BE0089) were also injected intraperitoneally. Subcutaneous
CO:COLLECTION_SUMMARY            	tumor growth was measured using calipers and thereafter tumor volume was
CO:COLLECTION_SUMMARY            	calculated using the formula for the volume of an ellipsoid given below. In the
CO:COLLECTION_SUMMARY            	case of intracranial tumors, tumor growth was measured using the luminescence
CO:COLLECTION_SUMMARY            	reader IVIS Ilumina and analyzed using LivingImage Software. Samples were
CO:COLLECTION_SUMMARY            	collected from mice with three biological replicates. Serum (50 µL) was
CO:COLLECTION_SUMMARY            	transferred to 200 μL ice-chilled (4°C) MilliQ H2O. Tissue (~16 mg) was
CO:COLLECTION_SUMMARY            	measured from subcutaneous and intracranial samples followed by addition of 250
CO:COLLECTION_SUMMARY            	μL MilliQ H2O. Then, samples were sonicated at 40 amps (~30 s) until
CO:COLLECTION_SUMMARY            	homogeneous. 80 μL of at 0.150 µg/mL debrisoquine in 60% methanol (MeOH)/40%
CO:COLLECTION_SUMMARY            	water(aq) reagent was added. 500 μL chilled (-20°C) MeOH was added, vortexed
CO:COLLECTION_SUMMARY            	(med) and incubated 15 min on ice. 250 μL chilled (-20°C) Chloroform was
CO:COLLECTION_SUMMARY            	added, vortexed (high) and incubated 20 min in ice on rotating mixer. Mixture
CO:COLLECTION_SUMMARY            	was centrifuged (13,000x g) for 18 min at 4°C. 705 μL of hydrophilic upper
CO:COLLECTION_SUMMARY            	layer was aspirated and transferred to separate 1.5 mL microtubes, dried to
CO:COLLECTION_SUMMARY            	completion under N2 gas sample concentrator, and stored at -80°C until LC/MS
CO:COLLECTION_SUMMARY            	quantification of GSK126.
CO:SAMPLE_TYPE                   	Tumor cells
CO:COLLECTION_METHOD             	Tumor Tissue/Serum Extract
CO:COLLECTION_LOCATION           	Intracranial/Subcutaneous/Serum from Mice
CO:COLLECTION_FREQUENCY          	Timepoints
CO:COLLECTION_DURATION           	0, 2h, 6h and 10h
CO:VOLUMEORAMOUNT_COLLECTED      	Serum (50 µL)/Tumor (~16 mg)
CO:STORAGE_CONDITIONS            	-80℃
CO:COLLECTION_VIALS              	15 mL
CO:STORAGE_VIALS                 	1.7 mL
CO:COLLECTION_TUBE_TEMP          	On Ice
CO:ADDITIVES                     	Extraction reagent
#TREATMENT
TR:TREATMENT_SUMMARY             	Treatment details • The mice were euthanized 2h, 6h and10h after drug
TR:TREATMENT_SUMMARY             	treatment 27days after tumor injections and 12 days of continuous drug
TR:TREATMENT_SUMMARY             	treatment. • The mice injected with intracranial tumors weighed approximately
TR:TREATMENT_SUMMARY             	16g and received 100mg/kg dose of GSK126/ vehicle injected intra-peritoneally in
TR:TREATMENT_SUMMARY             	100uL volume. • The mice injected with subcutaneous tumors weighed
TR:TREATMENT_SUMMARY             	approximately 19g and received 100mg/kg dose of GSK 126/ vehicle injected
TR:TREATMENT_SUMMARY             	intra-peritoneally in 120uL volume. • For euthanasia, the mice were injected
TR:TREATMENT_SUMMARY             	with 200mg/kg ketamine (200uL/mouse). • Tissues harvested (frozen immediately)
TR:TREATMENT_SUMMARY             	include: Intracranial mice a) Brain bearing tumor b) Lumbar lymph node c) Serum
TR:TREATMENT_SUMMARY             	from cardiac bleed Subcutaneous mice a) Sub-cutaneous tumor b) Tumor draining
TR:TREATMENT_SUMMARY             	inguinal lymph node (DLN) c) Non- tumor draining inguinal lymph node (NDLN) d)
TR:TREATMENT_SUMMARY             	Serum from cardiac bleed Drug Preparation • Dissolved 1g of GSK 126 powder in
TR:TREATMENT_SUMMARY             	8.325mL DMSO to get a conc. of 120.12mg/mL, aliquot and store in -80C. •
TR:TREATMENT_SUMMARY             	Dilute the GSK126 stock in 7 parts of Cyclodextrin (2-hydroxy-beta-cyclodextrin)
TR:TREATMENT_SUMMARY             	to get a concentration of 15mg/mL of drug (done fresh everyday) and then inject
TR:TREATMENT_SUMMARY             	100uL/120uL per mouse as indicated above. • Cyclodextrin was prepared by
TR:TREATMENT_SUMMARY             	dissolving 45g in 100mL ddH20 as indicated by manufacturer’s description. •
TR:TREATMENT_SUMMARY             	For the vehicle control, dilute 1 part of DMSO in 7 parts of Cyclodextrin (done
TR:TREATMENT_SUMMARY             	fresh everyday) and then inject 100uL/120uL per mouse as indicated above.
TR:TREATMENT_PROTOCOL_ID         	Treatment
TR:TREATMENT                     	Dissolved 1g of GSK 126 powder in 8.325mL DMSO to get a conc. of 120.12mg/mL,
TR:TREATMENT                     	aliquot and store in -80C. • Dilute the GSK126 stock in 7 parts of
TR:TREATMENT                     	Cyclodextrin (2-hydroxy-beta-cyclodextrin) to get a concentration of 15mg/mL of
TR:TREATMENT                     	drug (done fresh everyday) and then inject 100uL/120uL per mouse
TR:TREATMENT_ROUTE               	Intra-peritoneal injection of GSK126 and/or vehicle
TR:TREATMENT_DOSE                	100mg/kg dose of GSK 126
TR:TREATMENT_DOSEVOLUME          	100mg/kg dose of GSK 126/120 uL
TR:TREATMENT_DOSEDURATION        	0, 2, 6, 10h
TR:TREATMENT_VEHICLE             	1 part of DMSO in 7 parts of Cyclodextrin (done fresh everyday) and then inject
TR:TREATMENT_VEHICLE             	100uL/120uL per mouse
TR:ANIMAL_ENDP_EUTHANASIA        	Mice were injected with 200mg/kg ketamine (200uL/mouse).
TR:ANIMAL_ENDP_TISSUE_COLL_LIST  	Intracranial mice a) Brain bearing tumor b) Lumbar lymph node c) Serum from
TR:ANIMAL_ENDP_TISSUE_COLL_LIST  	cardiac bleed Subcutaneous mice a) Sub-cutaneous tumor b) Tumor draining
TR:ANIMAL_ENDP_TISSUE_COLL_LIST  	inguinal lymph node (DLN) c) Non- tumor draining inguinal lymph node (NDLN) d)
TR:ANIMAL_ENDP_TISSUE_COLL_LIST  	Serum from cardiac bleed
TR:TREATMENT_COMPOUND            	GSK126
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Serum (50 µL) was transferred to 200 μL ice-chilled (4°C) MilliQ H2O. Tissue
SP:SAMPLEPREP_SUMMARY            	(~16 mg) was measured from subcutaneous and intracranial samples followed by
SP:SAMPLEPREP_SUMMARY            	addition of 250 μL MilliQ H2O. Then, samples were sonicated at 40 amps (~30 s)
SP:SAMPLEPREP_SUMMARY            	until homogeneous. 80 μL of at 0.150 µg/mL debrisoquine in 60% methanol
SP:SAMPLEPREP_SUMMARY            	(MeOH)/40% water(aq) reagent was added. 500 μL chilled (-20°C) MeOH was added,
SP:SAMPLEPREP_SUMMARY            	vortexed (med) and incubated 15 min on ice. 250 μL chilled (-20°C) Chloroform
SP:SAMPLEPREP_SUMMARY            	was added, vortexed (high) and incubated 20 min in ice on rotating mixer.
SP:SAMPLEPREP_SUMMARY            	Mixture was centrifuged (13,000x g) for 18 min at 4°C. 705 μL of hydrophilic
SP:SAMPLEPREP_SUMMARY            	upper layer was aspirated and transferred to separate 1.5 mL microtubes, dried
SP:SAMPLEPREP_SUMMARY            	to completion under N2 gas sample concentrator, and stored at -80°C
SP:SAMPLEPREP_PROTOCOL_ID        	Extraction
SP:PROCESSING_STORAGE_CONDITIONS 	-80℃
SP:EXTRACTION_METHOD             	LCMS Extraction
SP:EXTRACT_CLEANUP               	Bligh-Dyer biphasic separation, discard protein disk and dried to completion
SP:EXTRACT_CLEANUP               	under N2 gas
SP:SAMPLE_RESUSPENSION           	80 uL 60% MeOH (aq)
SP:SAMPLE_SPIKING                	Internal standard 0.150 µg/mL debrisoquine(IS) was added to each
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Samples were injected at 8 µL over an 8.3 min gradient on the AdvanceBio Glycan
CH:CHROMATOGRAPHY_SUMMARY        	Map 2.1 x 100 mm 2.7µm column at 35°C with a flow rate of 0.220 mL/min. The LC
CH:CHROMATOGRAPHY_SUMMARY        	gradient only utilized LC/MS grade reagents when preparing mobile phases, A
CH:CHROMATOGRAPHY_SUMMARY        	(88:12 H2O/acetonitrile (ACN) and B 90% ACN (aq). Both mobile phases were
CH:CHROMATOGRAPHY_SUMMARY        	composed with 10 mM ammonium acetate and titrated to pH 6.85 using formic acid
CH:CHROMATOGRAPHY_SUMMARY        	and ammonium hydroxide. The LC gradient was initially 100% B for 0.25 min and
CH:CHROMATOGRAPHY_SUMMARY        	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
CH:CHROMATOGRAPHY_SUMMARY        	min; held for 0.5 min; 15% B at 7 min; 100%B at 8.3 min followed by
CH:CHROMATOGRAPHY_SUMMARY        	equilibration for 1.2 min.
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Agilent 1290 Infinity II
CH:COLUMN_NAME                   	Agilent AdvanceBio Glycan Map 2.1 x 100 mm 2.7µm column
CH:FLOW_GRADIENT                 	The LC gradient was initially 100% B for 0.25 min and then ramped to 55% B at
CH:FLOW_GRADIENT                 	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;
CH:FLOW_GRADIENT                 	15% B at 7 min; 100%B at 8.3 min followed by equilibration for 1.2 min.
CH:FLOW_RATE                     	0.220 mL/min
CH:COLUMN_TEMPERATURE            	35°C
CH:SOLVENT_A                     	88:12 H2O/acetonitrile (ACN) with 10 mM ammonium acetate and titrated to pH 6.85
CH:SOLVENT_B                     	90% ACN (aq) with 10 mM ammonium acetate and titrated to pH 6.85
CH:COLUMN_PRESSURE               	600 bar
CH:INJECTION_TEMPERATURE         	4°C
CH:INTERNAL_STANDARD             	Continuous accurate mass correction was achieved by infusing proprietary Agilent
CH:INTERNAL_STANDARD             	Technologies API-TOF reference mass standard solution
CH:RETENTION_TIME                	2.5-2.7 min
CH:SAMPLE_INJECTION              	8 uL
CH:SAMPLING_CONE                 	2kV
CH:ANALYTICAL_TIME               	8.3 min
CH:CAPILLARY_VOLTAGE             	3 kV
CH:SHEATH_LIQUID                 	N2
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
AN:LABORATORY_NAME               	NOB Cancer Metabolism
AN:OPERATOR_NAME                 	Dowdy
AN:SOFTWARE_VERSION              	8.0
AN:DATA_FORMAT                   	.d.zip
#MS
MS:INSTRUMENT_NAME               	Agilent 6545
MS:INSTRUMENT_TYPE               	QTOF
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	MS/MS Using Masshunter Qtof Quant-My-Way 10.0 software, GSK126 was detected at
MS:MS_COMMENTS                   	elution time 2.6 min using precursor ion m/z 527.3129 and transition m/z
MS:MS_COMMENTS                   	375.2183 generated via N2 gas collision-induced fragmentation (CID) at a
MS:MS_COMMENTS                   	collision energy (CE) of 12 V. Internal standard (IS) debrisoquine detected at
MS:MS_COMMENTS                   	elution time of 2.5 mins with precursor m/z 176.1182 and transition m/z 134.0964
MS:MS_COMMENTS                   	generated at CE 12V.
MS:CAPILLARY_TEMPERATURE         	325°C
MS:CAPILLARY_VOLTAGE             	3kV
MS:COLLISION_ENERGY              	12V
MS:COLLISION_GAS                 	N2
MS:DRY_GAS_FLOW                  	9 L/min
MS:DRY_GAS_TEMP                  	250°C
MS:FRAGMENT_VOLTAGE              	100 V
MS:FRAGMENTATION_METHOD          	MS/MS
MS:ION_SOURCE_TEMPERATURE        	325°C
MS:MASS_ACCURACY                 	2 kDa
MS:SOURCE_TEMPERATURE            	325°C
MS:CDL_TEMPERATURE               	RT
MS:DESOLVATION_TEMPERATURE       	325°C
MS:NEBULIZER                     	45 psig
MS:OCTPOLE_VOLTAGE               	750V
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	ng
MS_METABOLITE_DATA_START
Samples	Blank_S14.d	blank_S22.d	blank_Sr62.d	BRN_g1 _Intra1_2h	BRN_g1 _Intra2_2h	BRN_g1 _Intra3_2h	BRN_g1 _Intra4_6h	BRN_g1 _Intra5_6h	BRN_g1 _Intra7_10h	BRN_g1 _Intra8_10h	BRN_ctrl_g2_Intra10_2h	BRN_ctrl_g2_Intra11_2h	BRN_ctrl_g2_Intra12_6h	BRN_ctrl_g2_Intra13_6h	BRN_ctrl_g2_Intra14_6h	BRN_ctrl_g2_Intra15_10h	BRN_ctrl_g2_Intra16_10h	SubCu1_DLN_g1_2h	SubCu2_DLN_g1_2h	SubCu3_DLN_g1_2h	SubCu4_DLN_g1_6h	SubCu5_DLN_g1_6h	SubCu6_DLN_g1_6h	SubCu7_DLN_g1_10	SubCu8_DLN_g1_10	SubCu9_DLN_ctrl_g2_2h_	SubCu10_DLN_ctrl_g2_2h	SubCu11_DLN_ctrl_g2_2h	SubCu12_DLN_ctrl_g2_6h	SubCu13_DLN_ctrl_g2_6h	SubCu15_DLN_ctrl_g2_10h	SubCu16_DLN_ctrl_g2_10h	LLN_g1 _Intra1_2h	LLN_g1 _Intra2_2h	LLN_g1 _Intra3_2h	LLN_g1 _Intra4_6h	LLN_g1 _Intra6_6h	LLN_g1 _Intra7_10h	LLN_g1 _Intra8_10h	LLN_ctrl_g2_Intra10_2h	LLN_ctrl_g2_Intra11_2h	LLN_ctrl_g2_Intra12_6h	LLN_ctrl_g2_Intra13_6h	LLN_ctrl_g2_Intra15_10h	LLN_ctrl_g2_Intra16_10h	LLN_ctrl_g2 _Intra9_2h	SubCu1_NLN_g1_2h	SubCu2_NLN_g1_2h	SubCu3_NLN_g1_2h	SubCu4_NLN_g1_6h	SubCu5_NLN_g1_6h	SubCu6_NLN_g1_6h	SubCu7_NLN_g1_10	SubCu8_NLN_g1_10	SubCu9_NLN_ctrl_g2_2h_	SubCu10_NLN_ctrl_g2_2h	SubCu11_NLN_ctrl_g2_2h	SubCu12_NLN_ctrl_g2_6h	SubCu13_NLN_ctrl_g2_6h	SubCu14_NLN_ctrl_g2_6h	SubCu15_NLN_ctrl_g2_10h	SubCu16_NLN_ctrl_g2_10h	QcBRN_S45.d	QcBRN_Sr91.d	QcBRN_Sr111.d	QcBRN_Sr65.d	QcLLN_L1002.d	QCpool g1 D_NLN_subcu_055.d	QCpool g1 D_NLN_subcu_081.d	QCpool g1 D_NLN_subcu_135.d	QCpool g1 D_NLN_subcu_012.d	QCpool g1 D_NLN_subcu_028.d	QCpool g1 D_NLN_subcu_037.d	QCpool g1 SubTum_038.d	QCpool g1 SubTum_056.d	QCpool g1 SubTum_082.d	QCpool g1 SubTum_136.d	QcSer_S24.d	QcSer_S44.d	SER_g1 _Subcutaneous1_2h	SER_g1 _Subcutaneous2_2h	SER_g1 _Subcutaneous3_2h	SER_g1 _Subcutaneous4_6h	SER_g1 _Subcutaneous5_6h	SER_g1 _Subcutaneous6_6h	SER_g1 _Subcutaneous7_10h	SER_g1 _Subcutaneous8_10h	SER_g1 _Intra1_2h	SER_g1 _Intra2_2h	SER_g1 _Intra3_2h	SER_g1 _Intra4_6h	SER_g1 _Intra7_10h	SER_g1 _Intra8_10h	SER_g1 _Intra5_6h	SER_g1 _Intra6_6h	SER_ctrl_g2_Subcutaneous10_2h	SER_ctrl_g2_Subcutaneous12_6h	SER_ctrl_g2_Subcutaneous13_6h	SER_ctrl_g2_Subcutaneous14_6h	SER_ctrl_g2_Subcutaneous15_10h	SER_ctrl_g2_Subcutaneous16_10h	SER_ctrl_g2_Intra10_2h	SER_ctrl_g2_Intra11_2h	SER_ctrl_g2_Intra12_6h	SER_ctrl_g2_Intra13_6h	SER_ctrl_g2_Intra14_6h	SER_ctrl_g2_Intra15_10h	SER_ctrl_g2_Intra16_10h	SER_ctrl_g2 _Intra9_2h	SubCu1_Tum_g1_2h	SubCu2_Tum_g1_2h	SubCu3_Tum_g1_2h	SubCu4_Tum_g1_6h	SubCu5_Tum_g1_6h	SubCu6_Tum_g1_6h	SubCu7_Tum_g1_10	SubCu8_Tum_g1_10	SubCu9_Tum_ctrl_g2_2h_	SubCu10_Tum_ctrl_g2_2h	SubCu11_Tum_ctrl_g2_2h	SubCu12_Tum_ctrl_g2_6h	SubCu13_Tum_ctrl_g2_6h	SubCu14_Tum_ctrl_g2_6h	SubCu15_Tum_ctrl_g2_10h	SubCu16_Tum_ctrl_g2_10h
Factors	Treatment:blank | Gsk Exposure Time (h):-	Treatment:blank | Gsk Exposure Time (h):-	Treatment:blank | Gsk Exposure Time (h):-	Treatment:GSK | Gsk Exposure Time (h):2.0	Treatment:GSK | Gsk Exposure Time (h):2.0	Treatment:GSK | Gsk Exposure Time (h):2.0	Treatment:GSK | Gsk Exposure Time (h):6.0	Treatment:GSK | Gsk Exposure Time (h):6.0	Treatment:GSK | Gsk Exposure Time (h):10.0	Treatment:GSK | Gsk Exposure Time (h):10.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:GSK | Gsk Exposure Time (h):2.00	Treatment:GSK | Gsk Exposure Time (h):2.00	Treatment:GSK | Gsk Exposure Time (h):2.00	Treatment:GSK | Gsk Exposure Time (h):6.00	Treatment:GSK | Gsk Exposure Time (h):6.00	Treatment:GSK | Gsk Exposure Time (h):6.00	Treatment:GSK | Gsk Exposure Time (h):10	Treatment:GSK | Gsk Exposure Time (h):10	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:GSK | Gsk Exposure Time (h):2.0	Treatment:GSK | Gsk Exposure Time (h):2.0	Treatment:GSK | Gsk Exposure Time (h):2.0	Treatment:GSK | Gsk Exposure Time (h):6.0	Treatment:GSK | Gsk Exposure Time (h):6.0	Treatment:GSK | Gsk Exposure Time (h):10.0	Treatment:GSK | Gsk Exposure Time (h):10.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:GSK | Gsk Exposure Time (h):2.00	Treatment:GSK | Gsk Exposure Time (h):2.00	Treatment:GSK | Gsk Exposure Time (h):2.00	Treatment:GSK | Gsk Exposure Time (h):6.00	Treatment:GSK | Gsk Exposure Time (h):6.00	Treatment:GSK | Gsk Exposure Time (h):6.00	Treatment:GSK | Gsk Exposure Time (h):10	Treatment:GSK | Gsk Exposure Time (h):10	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:QC | Gsk Exposure Time (h):-	Treatment:QC | Gsk Exposure Time (h):-	Treatment:QC | Gsk Exposure Time (h):-	Treatment:QC | Gsk Exposure Time (h):-	Treatment:QC | Gsk Exposure Time (h):-	Treatment:QC | Gsk Exposure Time (h):-	Treatment:QC | Gsk Exposure Time (h):-	Treatment:QC | Gsk Exposure Time (h):-	Treatment:QC | Gsk Exposure Time (h):-	Treatment:QC | Gsk Exposure Time (h):-	Treatment:QC | Gsk Exposure Time (h):-	Treatment:QC | Gsk Exposure Time (h):-	Treatment:QC | Gsk Exposure Time (h):-	Treatment:QC | Gsk Exposure Time (h):-	Treatment:QC | Gsk Exposure Time (h):-	Treatment:QC | Gsk Exposure Time (h):-	Treatment:QC | Gsk Exposure Time (h):-	Treatment:GSK | Gsk Exposure Time (h):2.0	Treatment:GSK | Gsk Exposure Time (h):2.0	Treatment:GSK | Gsk Exposure Time (h):2.0	Treatment:GSK | Gsk Exposure Time (h):6.0	Treatment:GSK | Gsk Exposure Time (h):6.0	Treatment:GSK | Gsk Exposure Time (h):6.0	Treatment:GSK | Gsk Exposure Time (h):10.0	Treatment:GSK | Gsk Exposure Time (h):10.0	Treatment:GSK | Gsk Exposure Time (h):2.0	Treatment:GSK | Gsk Exposure Time (h):2.0	Treatment:GSK | Gsk Exposure Time (h):2.0	Treatment:GSK | Gsk Exposure Time (h):6.0	Treatment:GSK | Gsk Exposure Time (h):10.0	Treatment:GSK | Gsk Exposure Time (h):10.0	Treatment:GSK | Gsk Exposure Time (h):6.0	Treatment:GSK | Gsk Exposure Time (h):6.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:Vehicle | Gsk Exposure Time (h):0.0	Treatment:GSK | Gsk Exposure Time (h):2.00	Treatment:GSK | Gsk Exposure Time (h):2.00	Treatment:GSK | Gsk Exposure Time (h):2.00	Treatment:GSK | Gsk Exposure Time (h):6.00	Treatment:GSK | Gsk Exposure Time (h):6.00	Treatment:GSK | Gsk Exposure Time (h):6.00	Treatment:GSK | Gsk Exposure Time (h):10	Treatment:GSK | Gsk Exposure Time (h):10	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00	Treatment:Vehicle | Gsk Exposure Time (h):0.00
GSK126	0	0	0	0	41.768	25.344	0	13.712	13.744	0	0	0	0	0	0	0	0	126.35	449.75	677.39	2245.46	765.45	530.67	502.53	746.97	0	0	0	0	0	0	0	311.784	605.704	156.192	31.888	20.512	22.808	42.184	0	0	0	0	0	0	0	189.42	255.78	295.4	1089.55	415.1	553.98	1230.67	459.97	0	0	0	0	0	0	0	0	0	0	0	0	69.28	634.83	637.42	640.01	605.64	597.24	615.65	361.27	453.39	354.97	416.29	41.216	45.976	72.528	22.848	91.624	24.312	46.088	55.04	0	32	160.768	220.832	149.024	0	41.792	14.352	147.568	15.256	0	0	0	0	0	0	0	0	0	0	0	0	0	0	694.4	0	248.36	267.19	225.05	1329.09	107.24	223.72	0	0	0	0	0	0	0	0
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	retention index	quantified m/z	PubChem ID	KEGG ID
GSK126		527.3129	68210102	K11430
METABOLITES_END
#END