Summary of Study ST003281

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 PR002033. The data can be accessed directly via it's Project DOI: 10.21228/M8TN7J This work is supported by NIH grant, U2C- DK119886.

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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.

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Study IDST003281
Study TitlePhosphate availability conditions caspofungin tolerance, capsule attachment and titan cell formation in Cryptococcus neoformans
Study TypeMetabolomics and lipidomics
Study SummaryThere is a pressing need for new antifungal drugs to treat invasive fungal diseases. Unfortunately, the echinocandin drugs that are fungicidal against other important fungal pathogens are ineffective against Cryptococcus neoformans, the causative agent of life-threatening meningoencephalitis in immunocompromised people. Contributing mechanisms for echinocandin tolerance are emerging with connections to calcineurin signaling, the cell wall, and membrane composition. In this context, we discovered that a defect in phosphate uptake impairs the tolerance of C. neoformans to the echinocandin caspofungin.
Institute
University of British Columbia
DepartmentLife Sciences Institute
Last NameAlcazar Magana
First NameArmando
Address2350 Health Sciences Mall
Emailarmando.alcazarmagana@ubc.ca
Phone5416097172
Submit Date2024-06-12
Num Groups8
Total Subjects28
Raw Data AvailableYes
Raw Data File Type(s)mzML
Analysis Type DetailLC-MS
Release Date2024-06-28
Release Version1
Armando Alcazar Magana Armando Alcazar Magana
https://dx.doi.org/10.21228/M8TN7J
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR002033
Project DOI:doi: 10.21228/M8TN7J
Project Title:Phosphate availability conditions caspofungin tolerance, capsule attachment and titan cell formation in Cryptococcus neoformans
Project Summary:There is a pressing need for new antifungal drugs to treat invasive fungal diseases. Unfortunately, the echinocandin drugs that are fungicidal against other important fungal pathogens are ineffective against Cryptococcus neoformans, the causative agent of life-threatening meningoencephalitis in immunocompromised people. Contributing mechanisms for echinocandin tolerance are emerging with connections to calcineurin signaling, the cell wall, and membrane composition. In this context, we discovered that a defect in phosphate uptake impairs the tolerance of C. neoformans to the echinocandin caspofungin.
Institute:Life Sciences Institute, The University of British Columbia
Last Name:Alcazar Magana
First Name:Armando
Address:2350 Health Sciences Mall
Email:armando.alcazarmagana@ubc.ca
Phone:5416097172

Subject:

Subject ID:SU003401
Subject Type:Fungi
Subject Species:Cryptococcus neoformans

Factors:

Subject type: Fungi; Subject species: Cryptococcus neoformans (Factor headings shown in green)

mb_sample_id local_sample_id Genotype Treatment Sample source
SA355149H99-0_AH99 0 Cells
SA355150H99-0_BH99 0 Cells
SA355151H99-0_CH99 0 Cells
SA355152H99-250_CH99 250 Cells
SA355153H99-250_BH99 250 Cells
SA355154H99-250_AH99 250 Cells
SA355155H99-iron-20Pi-24h_AH99 H99-iron-20Pi-24h Cells
SA355156H99-iron-20Pi-24h_DH99 H99-iron-20Pi-24h Cells
SA355157H99-iron-20Pi-24h_CH99 H99-iron-20Pi-24h Cells
SA355158H99-iron-20Pi-24h_BH99 H99-iron-20Pi-24h Cells
SA355159H99-iron-20Pi-6h_AH99 H99-iron-20Pi-6h Cells
SA355160H99-iron-20Pi-6h_BH99 H99-iron-20Pi-6h Cells
SA355161H99-iron-20Pi-6h_CH99 H99-iron-20Pi-6h Cells
SA355162H99-iron-20Pi-6h_DH99 H99-iron-20Pi-6h Cells
SA355163H99-iron-24h_AH99 H99-iron-24h Cells
SA355164H99-iron-24h_DH99 H99-iron-24h Cells
SA355165H99-iron-24h_CH99 H99-iron-24h Cells
SA355166H99-iron-24h_BH99 H99-iron-24h Cells
SA355167H99-iron-6h_DH99 H99-iron-6h Cells
SA355168H99-iron-6h_CH99 H99-iron-6h Cells
SA355169H99-iron-6h_BH99 H99-iron-6h Cells
SA355170H99-iron-6h_AH99 H99-iron-6h Cells
SA355171H99-no iron-20Pi-6h_CH99 H99-no iron-20Pi-6h Cells
SA355172H99-no iron-20Pi-6h_DH99 H99-no iron-20Pi-6h Cells
SA355173H99-no iron-20Pi-6h_AH99 H99-no iron-20Pi-6h Cells
SA355174H99-no iron-20Pi-6h_BH99 H99-no iron-20Pi-6h Cells
SA355175H99-no iron-6h_BH99 H99-no iron-6h Cells
SA355176H99-no iron-6h_DH99 H99-no iron-6h Cells
SA355177H99-no iron-6h_AH99 H99-no iron-6h Cells
SA355178H99-no iron-6h_CH99 H99-no iron-6h Cells
SA355179QC3H99 QC Cells
SA355180QC5H99 QC Cells
SA355181QC4H99 QC Cells
SA355182QC1H99 QC Cells
SA355183QC2H99 QC Cells
SA355184Pho-0_APho 0 Cells
SA355185Pho-0_BPho 0 Cells
SA355186Pho-0_CPho 0 Cells
SA355187Pho-250_APho 250 Cells
SA355188Pho-250_BPho 250 Cells
SA355189Pho-250_CPho 250 Cells
SA355190QCL3QC QC Cells
SA355191QCL1QC QC Cells
SA355192QCL2QC QC Cells
Showing results 1 to 44 of 44

Collection:

Collection ID:CO003394
Collection Summary:The analysis was performed with cells grown overnight in YPD, transferred to MM at 30°C, and normalized to an OD600 of 2 with 0 mM or 250 mM Pi. After 24 h of incubation, 2 ml of cells were transferred (normalized to an OD600 of 2) into a 2 mL microcentrifuge tube. For metabolomics, cells were grown overnight in YPD, transferred into LIM (with 2.5 mM Pi) with iron (using dH2O instead of low iron water) for 24 h. 5х107 cells were transferred into LIM (low iron, low Pi), LIM (low iron, 20 mM Pi), LIM (iron, low Pi) and LIM (iron, 20 mM Pi) for 6h. 3.5х107 cells were collected into a 2 mL microcentrifuge tube. Cells were centrifuged at 13,000 rpm, 4oC for 10 min, washed three times with ice-cold nanopure water. Lipid extraction was performed using a biphasic system of cold methanol, methyltert-butyl ether (MTBE), and H2O, as described with some modifications (Matyash, et al., 2008)
Sample Type:Fungal cells

Treatment:

Treatment ID:TR003410
Treatment Summary:The analysis was performed with cells grown overnight in YPD, transferred to MM at 30°C, and normalized to an OD600 of 2 with 0 mM or 250 mM Pi.

Sample Preparation:

Sampleprep ID:SP003408
Sampleprep Summary:Lipid extraction was performed using a biphasic system of cold methanol, methyltert-butyl ether (MTBE), and H2O, as described with some modifications (Matyash, et al., 2008).

Combined analysis:

Analysis ID AN005375 AN005376
Analysis type MS MS
Chromatography type HILIC Reversed phase
Chromatography system Bruker Impact II Bruker Impact II
Column Agilent InfinityLab Poroshell 120 HILIC-Z column (2.7 µm particle size, 150 x 2.1 mm) Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um)
MS Type ESI ESI
MS instrument type QTOF QTOF
MS instrument name Bruker Impact HD Bruker Impact HD
Ion Mode NEGATIVE NEGATIVE
Units Rel Abundance Rel Abundance

Chromatography:

Chromatography ID:CH004074
Chromatography Summary:A novel method optimized for highly polar compounds was used to analyze samples using Hydrophilic Interaction Liquid Chromatography (HILIC)
Instrument Name:Bruker Impact II
Column Name:Agilent InfinityLab Poroshell 120 HILIC-Z column (2.7 µm particle size, 150 x 2.1 mm)
Column Temperature:30
Flow Gradient:the initial condition of 90% B was held for 2 minutes, followed by a linear gradient to 40% B over 6 minutes, then maintained at 40% B for 2 minutes, returned to 90% B over 1.1 minutes, and equilibrated for 4.9 minutes, resulting in a total run time of 16 minutes
Flow Rate:0.3 ml/min
Solvent A:100% water; 0.1% formic acid; 10 mM ammonium acetate; 5 µM medronic acid
Solvent B:90% acetonitrile/10% water ; 0.1% formic acid; 10 mM ammonium acetate; 5 µM medronic acid
Chromatography Type:HILIC
  
Chromatography ID:CH004075
Chromatography Summary:Separation of compounds was achieved using a multigradient method on an Acquity CSH
Instrument Name:Bruker Impact II
Column Name:Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um)
Column Temperature:65
Flow Gradient:0 min 15% B; 0–2 min 30% B; 2–2.5 min 50% B; 2.5–12 min 80% B; 12–12.5 min 99% B; 12.5–13.5 min 99% B; 13.5–13.7 min 15% B; 13.7-17 min 15% B.
Flow Rate:0.5 ml/min
Solvent A:60% acetonitrile/40% water; 0.1% formic acid; 10 mM ammonium formate
Solvent B:90% isopropanol/10% acetonitrile; 0.1% formic acid; 10 mM ammonium formate
Chromatography Type:Reversed phase

MS:

MS ID:MS005104
Analysis ID:AN005375
Instrument Name:Bruker Impact HD
Instrument Type:QTOF
MS Type:ESI
MS Comments:Raw data processing of metabolites employed Progenesis QI™ (V3.0.7600.27622) software with the METLIN™ plugin (V1.0.7642.33805)
Ion Mode:NEGATIVE
  
MS ID:MS005105
Analysis ID:AN005376
Instrument Name:Bruker Impact HD
Instrument Type:QTOF
MS Type:ESI
MS Comments:Raw data processing of metabolites employed Progenesis QI™ (V3.0.7600.27622) software with the METLIN™ plugin (V1.0.7642.33805)
Ion Mode:NEGATIVE
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