Summary of Study ST001733
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001109. The data can be accessed directly via it's Project DOI: 10.21228/M8BX1P This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST001733 |
| 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 |
| maria.delgadodolset@beca.ceu.es | |
| Phone | +34 913724700 4665 |
| Submit Date | 2021-03-17 |
| Num Groups | 2 |
| Total Subjects | 22 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | d |
| Analysis Type Detail | LC-MS |
| Release Date | 2021-06-17 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001109 |
| Project DOI: | doi: 10.21228/M8BX1P |
| 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 |
Subject:
| Subject ID: | SU001810 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
Subject type: Human; Subject species: Homo sapiens (Factor headings shown in green)
| mb_sample_id | local_sample_id | group |
|---|---|---|
| SA162645 | POL_21 | allergic |
| SA162646 | POL_25 | allergic |
| SA162647 | POL_28 | allergic |
| SA162648 | POL_11 | allergic |
| SA162649 | POL_19 | allergic |
| SA162650 | POL_12 | allergic |
| SA162651 | POL_15 | allergic |
| SA162652 | POL_24 | allergic |
| SA162653 | POL_4 | non-allergic |
| SA162654 | POL_6 | non-allergic |
| SA162655 | POL_8 | non-allergic |
| SA162656 | POL_31 | non-allergic |
| SA162657 | POL_5 | non-allergic |
| SA162658 | POL_23 | non-allergic |
| SA162659 | POL_14 | non-allergic |
| SA162660 | POL_2 | non-allergic |
| SA162661 | POL_20 | non-allergic |
| SA162662 | POL_22 | non-allergic |
| SA162663 | POL_29 | non-allergic |
| Showing results 1 to 19 of 19 |
Collection:
| Collection ID: | CO001803 |
| 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 ID: | TR001823 |
| Treatment Summary: | Plasma from heparinized blood was collected using a Ficoll-Paque (GE Healthcare™) density gradient centrifugation. |
Sample Preparation:
| Sampleprep ID: | SP001816 |
| 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. |
Combined analysis:
| Analysis ID | AN002821 | AN002822 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Agilent 1200 | Agilent 1200 |
| Column | Discovery HS C18 (150 x 2.1mm,3.0um) | Discovery HS C18 (150 x 2.1mm,3.0um) |
| MS Type | ESI | ESI |
| MS instrument type | QTOF | QTOF |
| MS instrument name | Agilent 6520 QTOF | Agilent 6520 QTOF |
| Ion Mode | NEGATIVE | POSITIVE |
| Units | peak area | peak area |
Chromatography:
| Chromatography ID: | CH002086 |
| 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. |
| Instrument Name: | Agilent 1200 |
| Column Name: | Discovery HS C18 (150 x 2.1mm,3.0um) |
| Column Temperature: | 40ºC |
| Flow Rate: | 0.6 mL/min |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% acetonitrile; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS002615 |
| Analysis ID: | AN002821 |
| Instrument Name: | Agilent 6520 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | The capillary voltage was set at 4,000 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 TFA NH4 (119.0363) and HP-0921 (966.0007). 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. 698 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 429 signals. |
| Ion Mode: | NEGATIVE |
| MS ID: | MS002616 |
| Analysis ID: | AN002822 |
| Instrument Name: | Agilent 6520 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| 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. |
| Ion Mode: | POSITIVE |
