#METABOLOMICS WORKBENCH emilygill_20191023_135935_mwtab.txt DATATRACK_ID:1837 STUDY_ID:ST001281 ANALYSIS_ID:AN002122 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 Metabolomics CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Dionex Ultimate 3000 CH:COLUMN_NAME ACE Excel 2 C18-PFP (100 x 2.1mm, 2um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Metabolomics MS:MS_RESULTS_FILE ST001281_AN002122_Results.txt UNITS:Peak Height Has m/z:Yes Has RT:Yes RT units:Minutes #END