{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001733","ANALYSIS_ID":"AN002822","VERSION":"1","CREATED_ON":"March 25, 2021, 11:06 am"},

"PROJECT":{"PROJECT_TITLE":"LC-MS Nasal Polyp analysis","PROJECT_SUMMARY":"Analysis of human samples from patients with nasal polyps with and without allergy.","INSTITUTE":"CEMBIO","LAST_NAME":"Delgado Dolset","FIRST_NAME":"María Isabel","ADDRESS":"Urb. Montepríncipe s/n, Ctra. Boadilla del Monte km 5.3, Madrid, Madrid, 28668, Spain","EMAIL":"maria.delgadodolset@beca.ceu.es","PHONE":"+34 913724700 4665"},

"STUDY":{"STUDY_TITLE":"Understanding systemic and local inflammation induced by nasal polyposis: role of the allergic phenotype (part-I)","STUDY_SUMMARY":"Chronic rhinosinusitis with nasal polyps (CRSwNP) is characterized by persistent symptoms associated to the development of nasal polyps. To this day, the molecular mechanisms involved are still not well defined. However, it has been suggested that a sustained inflammation as allergy is involved in its onset. In this pilot study, we aimed to look into the effect of the allergic status of the patient and in their underlying mechanisms. To achieve this, we recruited 22 patients with CRSwNP and classified them in non-allergic and allergic using ImmunoCAP ISAC molecular diagnosis. Plasma samples were analyzed using liquid chromatography coupled to mass spectrometry (LC-MS). Subsequently, the identified changed metabolites from plasma that were commercially available were then analyzed by targeted analysis in some nasal polyps. Additionally, nasal polyp and mucosa tissue samples were examined for eosinophils and neutrophils. We found that 9 out of the 22 patients were sensitized to some aeroallergens (named as allergic). The other 13 patients had no sensitizations (non-allergic). Regarding metabolomics, we found that bilirubin, cortisol, lysophosphatidylcholines (LPCs) 16:0, 18:0 and 20:4 and lysophosphatidylinositol (LPI) 20:4, metabolites that are usually related to a sustained allergic inflammation, were unexpectedly increased in the plasma of non-allergic patients with CRSwNP compared to allergic patients with CRSwNP. LPC 16:0, LPC 18:0 and LPI 20:4 metabolites followed the same trend in the nasal polyp as they did in plasma. Comparison of nasal polyps with nasal mucosa tissue showed a significant increase in eosinophils (p < 0.001) and neutrophils (p < 0.01) in allergic patients with CRSwNP. There were also more eosinophils in the polyps of non-allergic patients with CRSwNP than in their nasal mucosa (p <0.01). The polyps from non-allergic patients with CRSwNP had less eosinophils than the polyps of allergic patients with CRSwNP (p < 0.05). Our data suggests that there is a systemic inflammatory response associated to CRSwNP in the absence of allergy, which could be accountable for the nasal polyp development. Allergic patients with CRSwNP presented a higher number of eosinophils located in nasal polyps suggesting that eosinophilia might be connected to the development of nasal polyps in these patients.","INSTITUTE":"CEMBIO","LAST_NAME":"Delgado Dolset","FIRST_NAME":"María Isabel","ADDRESS":"Urb. Montepríncipe s/n, Ctra. Boadilla del Monte km 5.3, Madrid, Madrid, 28668, Spain","EMAIL":"maria.delgadodolset@beca.ceu.es","PHONE":"+34 913724700 4665","NUM_GROUPS":"2","TOTAL_SUBJECTS":"22"},

"SUBJECT":{"SUBJECT_TYPE":"Human","SUBJECT_SPECIES":"Homo sapiens","TAXONOMY_ID":"9606"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"POL_11",
"Factors":{"group":"allergic"},
"Additional sample data":{"RAW_FILE_NAME":"POL_11.d"}
},
{
"Subject ID":"-",
"Sample ID":"POL_12",
"Factors":{"group":"allergic"},
"Additional sample data":{"RAW_FILE_NAME":"POL_12.d"}
},
{
"Subject ID":"-",
"Sample ID":"POL_14",
"Factors":{"group":"non-allergic"},
"Additional sample data":{"RAW_FILE_NAME":"POL_14.d"}
},
{
"Subject ID":"-",
"Sample ID":"POL_15",
"Factors":{"group":"allergic"},
"Additional sample data":{"RAW_FILE_NAME":"POL_15.d"}
},
{
"Subject ID":"-",
"Sample ID":"POL_19",
"Factors":{"group":"allergic"},
"Additional sample data":{"RAW_FILE_NAME":"POL_19.d"}
},
{
"Subject ID":"-",
"Sample ID":"POL_2",
"Factors":{"group":"non-allergic"},
"Additional sample data":{"RAW_FILE_NAME":"POL_2.d"}
},
{
"Subject ID":"-",
"Sample ID":"POL_20",
"Factors":{"group":"non-allergic"},
"Additional sample data":{"RAW_FILE_NAME":"POL_20.d"}
},
{
"Subject ID":"-",
"Sample ID":"POL_21",
"Factors":{"group":"allergic"},
"Additional sample data":{"RAW_FILE_NAME":"POL_21.d"}
},
{
"Subject ID":"-",
"Sample ID":"POL_22",
"Factors":{"group":"non-allergic"},
"Additional sample data":{"RAW_FILE_NAME":"POL_22.d"}
},
{
"Subject ID":"-",
"Sample ID":"POL_23",
"Factors":{"group":"non-allergic"},
"Additional sample data":{"RAW_FILE_NAME":"POL_23.d"}
},
{
"Subject ID":"-",
"Sample ID":"POL_24",
"Factors":{"group":"allergic"},
"Additional sample data":{"RAW_FILE_NAME":"POL_24.d"}
},
{
"Subject ID":"-",
"Sample ID":"POL_25",
"Factors":{"group":"allergic"},
"Additional sample data":{"RAW_FILE_NAME":"POL_25.d"}
},
{
"Subject ID":"-",
"Sample ID":"POL_28",
"Factors":{"group":"allergic"},
"Additional sample data":{"RAW_FILE_NAME":"POL_28.d"}
},
{
"Subject ID":"-",
"Sample ID":"POL_29",
"Factors":{"group":"non-allergic"},
"Additional sample data":{"RAW_FILE_NAME":"POL_29.d"}
},
{
"Subject ID":"-",
"Sample ID":"POL_31",
"Factors":{"group":"non-allergic"},
"Additional sample data":{"RAW_FILE_NAME":"POL_31.d"}
},
{
"Subject ID":"-",
"Sample ID":"POL_4",
"Factors":{"group":"non-allergic"},
"Additional sample data":{"RAW_FILE_NAME":"POL_4.d"}
},
{
"Subject ID":"-",
"Sample ID":"POL_5",
"Factors":{"group":"non-allergic"},
"Additional sample data":{"RAW_FILE_NAME":"POL_5.d"}
},
{
"Subject ID":"-",
"Sample ID":"POL_6",
"Factors":{"group":"non-allergic"},
"Additional sample data":{"RAW_FILE_NAME":"POL_6.d"}
},
{
"Subject ID":"-",
"Sample ID":"POL_8",
"Factors":{"group":"non-allergic"},
"Additional sample data":{"RAW_FILE_NAME":"POL_8.d"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"We obtained 20 ml of heparinized blood from 19 out of the 22 patients. We used a Ficoll-Paque (GE Healthcare™) density gradient centrifugation to obtain plasma. Plasma samples were stored at -80ºC.","SAMPLE_TYPE":"Blood (plasma)","STORAGE_CONDITIONS":"-80℃"},

"TREATMENT":{"TREATMENT_SUMMARY":"Plasma from heparinized blood was collected using a Ficoll-Paque (GE Healthcare™) density gradient centrifugation."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Plasma proteins were removed by adding 300 µL of cold (-20 ℃) methanol:ethanol (1:1) to 100 µL of sample. Samples were then vortex-mixed and stored on ice for 5 min. Supernatant containing the metabolites was separated from the pellet by centrifugation (16,000 × g for 20 min at 4 ℃), then put into a LC vial for analysis. Quality control sample (QC) was prepared by pooling equal volumes of plasma from each sample. QC followed the same procedure applied for the experimental samples and was analysed throughout the run to provide a measurement of system stability, performance and reproducibility. All samples were randomised before metabolite extraction and for the corresponding analytical run."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Compound separation was performed on an Agilent HPLC system (1200 series, Agilent Technologies, Waldbronn, Germany) equipped with a degasser, two binary pumps, and a thermostated auto sampler. A volume of 10 μL of sample were injected into a Discovery HS C18 column (2.1 mm × 150 mm, 3.0 μm; Supelco, Sigma Aldrich, Germany), with a guard column Discovery® HS C18 (2 cm × 2.1 mm, 3 μm; Supelco, Sigma Aldrich, Germany), both maintained at 40 ℃. The flow rate was set at 0.6 mL/min. The elution gradient involved a mobile phase consisting of: (A) 0.1% formic acid (FA) in water, and (B) 0.1% FA in acetonitrile. Initial conditions were set at 25% phase B, which increased to 95% phase B in 35 min; then, it was re-equilibrated for 1 min and finally held for 9 min in the initial conditions.","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Agilent 1200","COLUMN_NAME":"Discovery HS C18 column (2.1 mm × 150 mm, 3.0 μm)","FLOW_RATE":"0.6 mL/min","COLUMN_TEMPERATURE":"40ºC","SOLVENT_A":"0.1% formic acid in water","SOLVENT_B":"0.1% FA in 100% acetonitrile"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","LABORATORY_NAME":"CEMBIO"},

"MS":{"INSTRUMENT_NAME":"Agilent 6520 QTOF","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"The capillary voltage was set at 3,500 V. The drying gas flow rate was 10.5 L/min at 330 ℃ and gas nebulizer at 52 psi; fragmentor voltage was 175 V; skimmer and octopole radio frequency voltage (OCT RF Vpp) were set to 65 and 750 V. Data were collected in the centroid mode at a scan rate of 1.2 spectra per second. Mass spectrometry detection was performed in full scan from 100 to 1200 m/z. The reference m/z ions were purine (121.0508) and HP-0921 (922.0097). These masses were continuously infused into the system to allow constant mass correction. Samples were analysed in separate runs. Acquired data were cleaned of background noises and unrelated ions using MassHunter Profinder (B.06.00, Agilent Technologies) software. “Molecular feature extraction” and “Find by ion” algorithms were applied to reduce the size and complexity of data, and to improve the reliability in finding the features. 1654 chemical signals were obtained. Then, data was filtered, and only those features detected in >50% in QCs and with a Relative Standard Deviation (RSD) <30% in Qcs were kept, resulting in 535 signals.","MS_RESULTS_FILE":"ST001733_AN002822_Results.txt UNITS:peak area Has m/z:Yes Has RT:Yes RT units:Minutes"}

}