#METABOLOMICS WORKBENCH emilygill_20191023_135935_mwtab.txt DATATRACK_ID:1837 STUDY_ID:ST001281 ANALYSIS_ID:AN002125 PROJECT_ID:PR000866
VERSION             	1
CREATED_ON             	November 19, 2019, 10:16 am
#PROJECT
PR:PROJECT_TITLE                 	Transgenic Parkinson's Mice Following Immunotherapy
PR:PROJECT_SUMMARY               	An UHPLC-HRMS Metabolomics and Lipidomics Study of Stool from Transgenic
PR:PROJECT_SUMMARY               	Parkinson's disease Mice Following Immunotherapy. Parkinson’s disease (PD) is
PR:PROJECT_SUMMARY               	characterized by the loss of dopaminergic neurons in the substantia nigra pars
PR:PROJECT_SUMMARY               	compacta of the brain as well as degeneration of motor and non-motor circuitry.
PR:PROJECT_SUMMARY               	The cause of neuronal death is currently unknown, although chronic
PR:PROJECT_SUMMARY               	neuroinflammation, aggregated α-synuclein, mitochondrial dysfunction and
PR:PROJECT_SUMMARY               	oxidative stress have all been implicated. Gliosis has been shown to exacerbate
PR:PROJECT_SUMMARY               	neuroinflammation via secretion of pro-inflammatory cytokines, and there is a
PR:PROJECT_SUMMARY               	subsequent infiltration of T lymphocytes (T-cells), into the brain of PD
PR:PROJECT_SUMMARY               	patients. Using liquid chromatography-high resolution mass spectrometry
PR:PROJECT_SUMMARY               	(LC-HRMS), we have observed metabolomic changes in stool samples, thought to be
PR:PROJECT_SUMMARY               	associated with the potential disease-modifying effect of an immunotherapy
PR:PROJECT_SUMMARY               	administered to transgenic Parkinsonian (A53T) mice. Significant elevations
PR:PROJECT_SUMMARY               	(p<0.05) in metabolites associated with immune response (taurine, histamine and
PR:PROJECT_SUMMARY               	its methylated product, 3-methylhistamine) are identified as being higher in the
PR:PROJECT_SUMMARY               	mice undergoing immunotherapy. Furthermore, a reduction in triacylglycerols (TG)
PR:PROJECT_SUMMARY               	and diacylglyceols (DG) expression in stool following immunotherapy suggests a
PR:PROJECT_SUMMARY               	regulation of lipid breakdown or biosynthesis with the vaccine. These
PR:PROJECT_SUMMARY               	“omics” markers (among others reported in this article) along with weight
PR:PROJECT_SUMMARY               	gain and increased life expectancy suggest that the immunotherapy is positively
PR:PROJECT_SUMMARY               	modifying the disease state.
PR:INSTITUTE                     	UNIVERSITY OF FLORIDA
PR:DEPARTMENT                    	CHEMISTRY
PR:LAST_NAME                     	GILL
PR:FIRST_NAME                    	EMILY
PR:ADDRESS                       	BUCKMAN DRIVE, GAINESVILLE, FL, 32611, USA
PR:EMAIL                         	emilygill2014@ufl.edu
PR:PHONE                         	(352) 222-9749
PR:FUNDING_SOURCE                	Southeast Center for Intergrated Metabolomics
PR:PUBLICATIONS                  	J Proteome Res, 2019.
PR:CONTRIBUTORS                  	Emily L. Gill, Jeremy P. Koelmel, Laurel Meke, Richard A. Yost, Timothy J.
PR:CONTRIBUTORS                  	Garrett, Michael S. Okun, Catherine Flores, and Vinata Vedam-Mai
#STUDY
ST:STUDY_TITLE                   	Transgenic Parkinson's Mice Following Immunotherapy
ST:STUDY_SUMMARY                 	An UHPLC-HRMS Metabolomics and Lipidomics Study of Stool from Transgenic
ST:STUDY_SUMMARY                 	Parkinson's disease Mice Following Immunotherapy. Parkinson’s disease (PD) is
ST:STUDY_SUMMARY                 	characterized by the loss of dopaminergic neurons in the substantia nigra pars
ST:STUDY_SUMMARY                 	compacta of the brain as well as degeneration of motor and non-motor circuitry.
ST:STUDY_SUMMARY                 	The cause of neuronal death is currently unknown, although chronic
ST:STUDY_SUMMARY                 	neuroinflammation, aggregated α-synuclein, mitochondrial dysfunction and
ST:STUDY_SUMMARY                 	oxidative stress have all been implicated. Gliosis has been shown to exacerbate
ST:STUDY_SUMMARY                 	neuroinflammation via secretion of pro-inflammatory cytokines, and there is a
ST:STUDY_SUMMARY                 	subsequent infiltration of T lymphocytes (T-cells), into the brain of PD
ST:STUDY_SUMMARY                 	patients. Using liquid chromatography-high resolution mass spectrometry
ST:STUDY_SUMMARY                 	(LC-HRMS), we have observed metabolomic changes in stool samples, thought to be
ST:STUDY_SUMMARY                 	associated with the potential disease-modifying effect of an immunotherapy
ST:STUDY_SUMMARY                 	administered to transgenic Parkinsonian (A53T) mice. Significant elevations
ST:STUDY_SUMMARY                 	(p<0.05) in metabolites associated with immune response (taurine, histamine and
ST:STUDY_SUMMARY                 	its methylated product, 3-methylhistamine) are identified as being higher in the
ST:STUDY_SUMMARY                 	mice undergoing immunotherapy. Furthermore, a reduction in triacylglycerols (TG)
ST:STUDY_SUMMARY                 	and diacylglyceols (DG) expression in stool following immunotherapy suggests a
ST:STUDY_SUMMARY                 	regulation of lipid breakdown or biosynthesis with the vaccine. These
ST:STUDY_SUMMARY                 	“omics” markers (among others reported in this article) along with weight
ST:STUDY_SUMMARY                 	gain and increased life expectancy suggest that the immunotherapy is positively
ST:STUDY_SUMMARY                 	modifying the disease state.
ST:INSTITUTE                     	UNIVERSITY OF FLORIDA
ST:DEPARTMENT                    	CHEMISTRY
ST:LAST_NAME                     	GILL
ST:FIRST_NAME                    	EMILY
ST:ADDRESS                       	BUCKMAN DRIVE, GAINESVILLE,FL ,32611, USA
ST:EMAIL                         	emilygill2014@ufl.edu
ST:PHONE                         	(352) 222-9749
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS           	-	Tg1	Treatment:Control	
SUBJECT_SAMPLE_FACTORS           	-	Tg2	Treatment:Control	
SUBJECT_SAMPLE_FACTORS           	-	Tg3	Treatment:Control	
SUBJECT_SAMPLE_FACTORS           	-	T1	Treatment:Vaccinated	
SUBJECT_SAMPLE_FACTORS           	-	T2	Treatment:Vaccinated	
SUBJECT_SAMPLE_FACTORS           	-	T3	Treatment:Vaccinated	
SUBJECT_SAMPLE_FACTORS           	-	Blank	Treatment:Blank	
#COLLECTION
CO:COLLECTION_SUMMARY            	Stool samples were collected at 12 months, while the mice were under clinical
CO:COLLECTION_SUMMARY            	observation. Stool was the chosen sample of analysis because of its ease of
CO:COLLECTION_SUMMARY            	collection.
CO:SAMPLE_TYPE                   	Feces
CO:STORAGE_CONDITIONS            	-80℃
#TREATMENT
TR:TREATMENT_SUMMARY             	All animal procedures were approved by UF IACUC, at the University of Florida.
TR:TREATMENT_SUMMARY             	Animals were housed in a 12h light-12h dark schedule, and were provided with a
TR:TREATMENT_SUMMARY             	diet of Envigo Mouse Chow (Huntingdon, England) and water ad libitum. Animals
TR:TREATMENT_SUMMARY             	were housed in groups, including 3 PD and 3 vaccinated PD mice. Female,
TR:TREATMENT_SUMMARY             	age-matched heterozygous M83 mice were used in all experiments. The
TR:TREATMENT_SUMMARY             	immunotherapy was administered at 6 months, prior to the development of any
TR:TREATMENT_SUMMARY             	overt physical symptoms of PD. Briefly, dendritic cells (DC) were pulsed with
TR:TREATMENT_SUMMARY             	α-syn (A53T) RNA to generate DC vaccine, and A53T α-syn-specific T cells were
TR:TREATMENT_SUMMARY             	generated ex vivo for adoptive transfer into M83 mice expressing the α-syn
TR:TREATMENT_SUMMARY             	mutation. A53T α-syn-specific T cells were adoptively transferred intravenously
TR:TREATMENT_SUMMARY             	after 5-7d of co-culture via a single injection of 107 autologous ex
TR:TREATMENT_SUMMARY             	vivo–expanded T cells.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Folch extraction for lipidomics and protein precipitation for metabolomics.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Lipidomics
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Thermo Dionex Ultimate 3000
CH:COLUMN_NAME                   	Waters Acquity BEH C18 (50 x 2.1mm, 1.7 um)
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	Lipidomics
MS:MS_RESULTS_FILE               	ST001281_AN002125_Results.txt	UNITS:Peak Height	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#END