#METABOLOMICS WORKBENCH jwalejko_20200818_081055 DATATRACK_ID:2123 STUDY_ID:ST001448 ANALYSIS_ID:AN002419 PROJECT_ID:PR000995
VERSION             	1
CREATED_ON             	August 20, 2020, 11:40 am
#PROJECT
PR:PROJECT_TITLE                 	Maternal Hypercortisolemia alters placental metabolism
PR:PROJECT_SUMMARY               	Previous studies have suggested that increases in maternal cortisol or maternal
PR:PROJECT_SUMMARY               	stress in late pregnancy increase the risk of stillbirth at term. In an ovine
PR:PROJECT_SUMMARY               	model with increased maternal cortisol over the last 0.20 of gestation, we have
PR:PROJECT_SUMMARY               	previously found evidence of disruption of fetal serum and cardiac metabolomics,
PR:PROJECT_SUMMARY               	and altered expression of genes related to mitochondrial function and metabolism
PR:PROJECT_SUMMARY               	in biceps femoris, diaphragm and cardiac muscle. The present studies were
PR:PROJECT_SUMMARY               	designed to test for effects of chronically increased maternal cortisol on gene
PR:PROJECT_SUMMARY               	expression and metabolomics in placentomes near term. We hypothesized that
PR:PROJECT_SUMMARY               	changes in placenta may underlie or contribute to the alterations in fetal serum
PR:PROJECT_SUMMARY               	metabolomics, and thereby contribute to changes in striated muscle metabolism.
PR:PROJECT_SUMMARY               	Placentomes were collected from pregnancies in early labor (143±1 d gestation)
PR:PROJECT_SUMMARY               	of control ewes (n=7) or ewes treated with cortisol (1 mg/kg/d iv; n=5) starting
PR:PROJECT_SUMMARY               	at day 115 of gestation. Transcriptomics and metabolomics were performed using
PR:PROJECT_SUMMARY               	an ovine gene expression microarray (Agilent 019921) and HR-MAS NMR,
PR:PROJECT_SUMMARY               	respectively. Multi-omic analysis indicates that amino acid metabolism,
PR:PROJECT_SUMMARY               	particularly of branched chain amino acids and glutamate occur in placenta;
PR:PROJECT_SUMMARY               	changes in amino acid metabolism, degradation or biosynthesis in placenta were
PR:PROJECT_SUMMARY               	consistent with changes in valine, isoleucine, leucine and glycine in fetal
PR:PROJECT_SUMMARY               	serum. The analysis also indicates changes in glycerophospholipid metabolism and
PR:PROJECT_SUMMARY               	suggests changes in ER stress and antioxidant status in the placenta. These
PR:PROJECT_SUMMARY               	findings suggest that changes in placental function occurring with excess
PR:PROJECT_SUMMARY               	maternal cortisol in late gestation may contribute to metabolic dysfunction at
PR:PROJECT_SUMMARY               	birth.
PR:INSTITUTE                     	University of Florida
PR:DEPARTMENT                    	Pharmacodynamics
PR:LAST_NAME                     	Keller-Wood
PR:FIRST_NAME                    	Maureen
PR:ADDRESS                       	1345 SW Archer Rd, PO 100487, Gainesville, FL 32610
PR:EMAIL                         	kellerwd@cop.ufl.edu
PR:PHONE                         	352-273-7687
#STUDY
ST:STUDY_TITLE                   	Maternal Hypercortisolemia alters placental metabolism: NMR metabolomics
ST:STUDY_SUMMARY                 	NMR metabolomic studies of placental tissue from sheep with excess maternal
ST:STUDY_SUMMARY                 	cortisol during late gestation
ST:INSTITUTE                     	University of Florida
ST:LAST_NAME                     	Walejko
ST:FIRST_NAME                    	Jacquelyn
ST:ADDRESS                       	300 N Duke St Durham NC 27701
ST:EMAIL                         	jacquelyn.walejko@duke.edu
ST:PHONE                         	9194792304
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Ovis aries
SU:TAXONOMY_ID                   	9940
#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           	-	656	Treatment:Cortisol | Sex:Female	RAW_FILE_NAME=1
SUBJECT_SAMPLE_FACTORS           	-	784	Treatment:Control | Sex:Male	RAW_FILE_NAME=3
SUBJECT_SAMPLE_FACTORS           	-	665	Treatment:Cortisol | Sex:Female	RAW_FILE_NAME=5
SUBJECT_SAMPLE_FACTORS           	-	749	Treatment:Control | Sex:Female	RAW_FILE_NAME=6
SUBJECT_SAMPLE_FACTORS           	-	521	Treatment:Control | Sex:Male	RAW_FILE_NAME=7
SUBJECT_SAMPLE_FACTORS           	-	874	Treatment:Cortisol | Sex:Female	RAW_FILE_NAME=8
SUBJECT_SAMPLE_FACTORS           	-	993A	Treatment:Control | Sex:Male	RAW_FILE_NAME=9
SUBJECT_SAMPLE_FACTORS           	-	713	Treatment:Cortisol | Sex:Female	RAW_FILE_NAME=10
SUBJECT_SAMPLE_FACTORS           	-	993B	Treatment:Control | Sex:Female	RAW_FILE_NAME=11
SUBJECT_SAMPLE_FACTORS           	-	568	Treatment:Control | Sex:Female	RAW_FILE_NAME=12
SUBJECT_SAMPLE_FACTORS           	-	594	Treatment:Control | Sex:Male	RAW_FILE_NAME=13
SUBJECT_SAMPLE_FACTORS           	-	768	Treatment:Cortisol | Sex:Female	RAW_FILE_NAME=14
#COLLECTION
CO:COLLECTION_SUMMARY            	Placentomes were collected at necropsy from control ewes and ewes treated with
CO:COLLECTION_SUMMARY            	cortisol (CORT: 1mg/kg/d from day 115 of pregnancy) at days 141-144 of
CO:COLLECTION_SUMMARY            	gestation. CORT ewes received an intravenous infusion of cortisol (Solu-Cortef,
CO:COLLECTION_SUMMARY            	hydrocortisone sodium succinate in sodium phosphate; Pfizer, New York, NY);
CO:COLLECTION_SUMMARY            	control animals did not receive any treatment. Animals were sacrificed for
CO:COLLECTION_SUMMARY            	collection of tissues at the first signs of labor, as evidenced by changes in
CO:COLLECTION_SUMMARY            	uterine blood flow; placentomes were collected from sheep in early stages of
CO:COLLECTION_SUMMARY            	labor at between 141-144 days and 142-144 days of gestation in the
CO:COLLECTION_SUMMARY            	cortisol-treated and control groups, respectively. In this cohort of ewes, there
CO:COLLECTION_SUMMARY            	were 7 control ewes, of which data were collected from placentomes of 7 live
CO:COLLECTION_SUMMARY            	fetuses (including one set of twins); there was one lamb born, and that placenta
CO:COLLECTION_SUMMARY            	was not collected. In this cohort there were 11 cortisol- treated ewes, from
CO:COLLECTION_SUMMARY            	which we collected placentomes from 5 live fetuses. There was one live lamb
CO:COLLECTION_SUMMARY            	born, and 5 stillborn lambs, from which placentas were not collected.
CO:COLLECTION_SUMMARY            	Placentomes were classified as A-D placentomes, snap frozen in liquid nitrogen,
CO:COLLECTION_SUMMARY            	and stored in -80C until metabolomic analysis.
CO:SAMPLE_TYPE                   	Placenta
#TREATMENT
TR:TREATMENT_SUMMARY             	Placentomes were collected at necropsy from control ewes and ewes treated with
TR:TREATMENT_SUMMARY             	cortisol (CORT: 1mg/kg/d from day 115 of pregnancy) at days 141-144 of
TR:TREATMENT_SUMMARY             	gestation. CORT ewes received an intravenous infusion of cortisol (Solu-Cortef,
TR:TREATMENT_SUMMARY             	hydrocortisone sodium succinate in sodium phosphate; Pfizer, New York, NY);
TR:TREATMENT_SUMMARY             	control animals did not receive any treatment.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	High-resolution magic angle spinning (HR-MAS) proton nuclear magnetic resonance
SP:SAMPLEPREP_SUMMARY            	(1H-NMR) was used to determine metabolites present in placental tissue. Briefly,
SP:SAMPLEPREP_SUMMARY            	30 μL of deuterium oxide (D2O) with 3.3 mM 3-(Trimethylsilyl)-1-propanesulfonic
SP:SAMPLEPREP_SUMMARY            	acid (DSS) as a reference standard was added to 27.7-52.1 mg (mean 35.3 mg) of
SP:SAMPLEPREP_SUMMARY            	placental tissue before being placed into a 4 mm HR-MAS rotor (Bruker Biospin)
SP:SAMPLEPREP_SUMMARY            	and spun at 6 kHz with a spectral width of 10 ppm. Data were acquired on an
SP:SAMPLEPREP_SUMMARY            	Avance III 600 MHz Bruker NMR spectrometer equipped with a 4 mm HR-MAS probe at
SP:SAMPLEPREP_SUMMARY            	the University of Georgia Complex Carbohydrate Research Center. Data were
SP:SAMPLEPREP_SUMMARY            	acquired using a one-dimensional experiment with T2 filter using
SP:SAMPLEPREP_SUMMARY            	Carr-Purcell-Meiboom-Gill (CPMG) pulse sequence with water presaturation
SP:SAMPLEPREP_SUMMARY            	(CPMGPR1D).
#ANALYSIS
AN:ANALYSIS_TYPE                 	NMR
#NMR
NM:INSTRUMENT_NAME               	Bruker Avance III HD
NM:INSTRUMENT_TYPE               	FT-NMR
NM:NMR_EXPERIMENT_TYPE           	1D-1H
NM:SPECTROMETER_FREQUENCY        	600 MHz
#NMR_METABOLITE_DATA
NMR_METABOLITE_DATA:UNITS	Area Under Curve
NMR_METABOLITE_DATA_START
Samples	656	665	874	713	768	784	749	521	993A	993B	568	594
Factors	Treatment:Cortisol | Sex:Female	Treatment:Cortisol | Sex:Female	Treatment:Cortisol | Sex:Female	Treatment:Cortisol | Sex:Female	Treatment:Cortisol | Sex:Female	Treatment:Control | Sex:Male	Treatment:Control | Sex:Female	Treatment:Control | Sex:Male	Treatment:Control | Sex:Male	Treatment:Control | Sex:Female	Treatment:Control | Sex:Female	Treatment:Control | Sex:Male
Lipid (-CH3)	2.242389176	3.809119519	3.950549632	3.396744874	3.655016682	4.138817585	4.25989936	3.514435259	4.55458362	3.268513682	3.538174986	2.633201729
Leucine	4.299739627	2.285780203	2.565743146	2.577903522	2.180277276	2.263470942	2.311929583	1.931117806	2.311563857	2.931377311	1.832554758	2.273570102
Isoleucine	0.794372466	0.454519294	0.437997067	0.49692665	0.401696967	0.432044676	0.43948745	0.330387715	0.442449931	0.540842677	0.348272318	0.428864983
Valine	1.37102458	0.595426446	0.732510268	0.673351146	0.539095761	0.542604821	0.605476821	0.444890302	0.595956857	0.851294909	0.476916156	0.616046206
Lipid (-CH)	2.151627931	7.618917475	7.198638109	5.03204128	5.97616203	6.957224374	8.189074535	5.494842384	8.752739029	5.207249128	6.643386518	4.185869228
Alanine	2.345442355	1.644259554	1.597410534	2.01311568	1.559373932	1.706268296	1.807180903	1.287549471	1.669190904	2.030010551	1.754121876	1.6439098
Lipid (Beta-CH2)	0.744009241	1.140799275	1.183847165	0.742519474	0.991843252	1.181927832	1.427785203	0.667738618	1.184395545	1.013666337	1.324415431	0.792787305
Acetate	0.659579979	0.494082232	0.400918887	0.391565064	0.45627498	0.470601668	0.478647967	0.37631367	0.480458385	0.633795247	0.484801557	0.434949441
Glutamate	1.277005783	1.00378418	0.768801494	0.782834966	0.895942788	1.285719377	1.098331256	0.991273294	1.039629152	1.451644542	1.141807501	1.163723643
Glutamine	0.418230197	0.372575265	0.264231756	0.21465936	0.348815321	0.549392019	0.51483962	0.21620621	0.331322904	0.5745901	0.391412043	0.378787836
Glutathione	0.193648648	0.217998595	0.183238029	0.105833648	0.171155398	0.356070925	0.250237334	0.1991357	0.222959112	0.316220518	0.226829608	0.331995569
Methionine	0.026302857	0.019267512	0.014984792	0.008334918	0.007607154	0.038558538	0.04093619	-0.026593911	0.006017306	0.05254543	0.021600042	0.051010083
Aspartate	0.03904523	0.094370031	0.126118128	0.068019275	0.1017084	0.057611096	0.106786565	0.025635939	0.098884719	0.062016138	0.112649903	0.093672828
Asparagine	0.277818685	0.157796407	0.394412548	0.220016148	0.26643748	0.35146956	0.319615787	0.301808355	0.196738036	0.325908917	0.329475948	0.452363009
Lysine	0.458273233	0.195481383	0.361293805	0.321635425	0.31668478	0.428887475	0.412711996	0.310095938	0.213878465	0.44492525	0.343061966	0.454891857
Creatine	0.247824619	0.122777032	0.192405855	0.227370258	0.195691824	0.230859314	0.212638688	0.298822066	0.170363881	0.260054776	0.236886705	0.309058819
Citrulline	0.173371345	0.079198602	0.155735177	0.182221461	0.166235472	0.187712744	0.17835926	0.216001999	0.095668788	0.176071225	0.200864364	0.192748351
Ethanolamine	0.244289509	0.089812417	0.252444042	0.206457973	0.236097441	0.28122313	0.38891794	0.290953595	0.191569078	0.310790184	0.472176639	0.322156624
Choline/Phosphocholine	0.618561466	0.541288363	0.703739064	0.974814614	1.159084115	0.975135935	0.934245546	1.24277794	1.126000719	1.30116655	1.269952513	0.783616129
Glycerophosphocholine	2.443749854	3.219526726	3.970876212	3.52611345	4.76797191	4.453432287	3.920177866	5.464910377	3.074437278	3.316314477	3.69046827	3.074642108
Proline	0.048447217	0.049735477	0.047445648	0.05128176	0.044542256	0.05288908	0.055253819	0.063343132	0.045499847	0.057211642	0.040695459	0.059964736
Taurine	0.749618285	0.675089376	0.754965567	1.038565829	0.675809754	0.817921033	0.698680329	1.303274826	0.882079174	0.878619167	0.860958332	1.1332409
Glycine	1.683605721	1.870524114	1.784684872	2.097562025	2.350489154	2.159715145	2.042294406	2.610485925	2.46699209	2.162102118	2.208831717	1.950770385
Glycerol	0.231278767	0.162129213	0.208723823	0.22050995	0.179615066	0.148876694	0.193345036	0.233249231	0.187360323	0.166565049	0.180569094	0.206557889
Betaine	0.613755433	0.829675526	0.627234326	0.837961731	0.811402286	0.736253166	0.485531011	0.828994216	0.683175418	0.593758057	0.613635743	0.5880968
Serine	0.852330779	0.655197443	0.623871014	0.800257822	0.618749534	0.597283499	0.564267224	0.755890194	0.643671595	0.703642742	0.658714893	0.751270317
Myo-inositol	0.844819633	0.872252606	0.804992851	1.00412676	0.919143422	0.78846167	0.761147692	1.036530071	1.126417223	1.017673907	0.938111878	0.867510265
Lactate	2.732545944	2.429187895	2.561082699	4.471084327	2.425466251	2.407846583	2.505302188	2.66403392	2.982596509	2.785578646	2.416090008	2.904588835
Phosphorylcholine	1.341951725	2.195885888	1.706068139	1.585747377	2.04107288	1.874164685	1.729538573	2.116383272	1.751710247	1.555504842	1.534353621	1.316810155
Threonine	0.661472796	0.656890868	0.73883513	0.64251404	0.625810738	0.629719395	0.757254297	0.682132627	0.575230119	0.613952441	0.617802723	0.643376618
Ascorbate	0.227792617	0.295165904	0.261258425	0.253565982	0.23520622	0.320598814	0.287114603	0.410699774	0.299950677	0.263006707	0.281428084	0.324791655
Lipid (-CH=CH-)	0.493439074	1.698877297	2.391124903	1.260792208	1.739888775	2.10193387	2.373382	1.576506138	2.71863695	1.518887431	2.341554524	1.202448307
Fumarate	0.003642458	-0.00175148	0.003298625	0.008688803	0.003924634	0.002146943	0.001617812	0.008953854	0.004843811	0.005215251	0.00500807	0.009014798
1-Methyhistidine/Histidine	0.049217533	0.105155526	0.019304743	0.083801019	0.125307956	0.034629361	-0.000826305	0.037518108	0.058383749	0.054295987	0.018788935	0.039356566
Tyrosine	0.152698368	0.111743917	0.121381889	0.113832298	0.090401946	0.100100729	0.136878418	0.089862116	0.0935251	0.167243113	0.064844048	0.162604109
Phenylalanine	0.752031727	0.415012303	0.40537087	0.429767729	0.390555387	0.469763181	0.486214106	0.269391831	0.399798635	0.776721862	0.325755725	0.541910383
Uracil	0.043676492	0.038968046	0.028400809	0.040002165	0.04911261	0.04189195	0.045324677	0.036725013	0.05526046	0.060442039	0.042148192	0.043800303
Inosine	0.123765307	0.175609736	0.186814852	0.146646264	0.135934711	0.175452827	0.152331892	0.242812743	0.166712098	0.126423522	0.176440428	0.210683636
NMR_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	PubChem	KEGG
Lipid (-CH3)	-	-
Leucine	6106	C00123
Isoleucine	6306	C00407
Valine	6287	C00183
Lipid (-CH)	-	-
Alanine	5950	C00041
Lipid (Beta-CH2)	-	-
Acetate	176	C00033
Glutamate	33032	C00025
Glutamine	5961	C00064
Glutathione	3353	C00051
Methionine	6137	C00073
Aspartate	5960	C00049
Asparagine	6267	C00152
Lysine	5962	C00047
Creatine	586	C00300
Citrulline	9750	C00327
Ethanolamine	700	C00189
Choline/Phosphocholine	305/1014	C00114/C00588
Glycerophosphocholine	71920	C00670
Proline	145742	C00148
Taurine	1123	C00245
Glycine	750	C00037
Glycerol	753	C00116
Betaine	247	C00719
Serine	3982	C00716
Myo-inositol	-	C00137
Lactate	107689	C00256
Phosphorylcholine	1014	C00588
Threonine	6288	C00188
Ascorbate	3372	C00072
Lipid (-CH=CH-)	-	-
Fumarate	444972	C00122
1-Methyhistidine/Histidine	92105/6274	C01152/C00135
Tyrosine	6057	C00082
Phenylalanine	6140	C00079
Uracil	1174	C00106
Inosine	135398641	-
METABOLITES_END
#END