Summary of Study ST001922
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 PR001213. The data can be accessed directly via it's Project DOI: 10.21228/M8X70P This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST001922 |
| Study Title | Sublytic membrane attack complex drives glycolysis and mitochondrial dysfunction with inflammatory consequences in human monocyte-derived macrophages |
| Study Summary | The terminal stage in the complement activation pathways, the membrane attack complex (MAC), is upregulated in diabetic and rheumatoid arthritis patients, contributing pathologically by increasing inflammation. Previous research has highlighted that a sublytic dose of MAC can initiate NLRP3 inflammasome activation via calcium influx and loss of mitochondrial membrane potential. Here, we show that sublytic concentrations of MAC mediate a previously undescribed perturbation in cellular energy metabolism in human monocyte-derived macrophages, by phenotypic skewing towards glycolysis and upregulation of glycolysis-promoting genes. Sublytic MAC concentrations drive mitochondrial dysfunction, characterised by a fragmented mitochondrial morphology, loss of maximal respiratory response, depleted mitochondrial membrane potential as well as increased mitochondrial reactive oxygen species production. The consequences of these alterations in glycolytic metabolism and mitochondrial dysfunction lead to NLRP3 inflammasome activation, driving gasdermin D formation and IL-18 release. This novel link between sublytic MAC and immunometabolism, with direct consequences for downstream inflammatory processes, is important for development of novel therapeutics for areas where MAC may mediate disease. |
| Institute | GSK |
| Department | Discovery Analytical |
| Laboratory | MST-MedDesign |
| Last Name | Kozole |
| First Name | Joseph |
| Address | 1250 Collegeville Ave, Upper Providence, PA, US |
| joseph.x.kozole@gsk.com | |
| Phone | 8144410679 |
| Submit Date | 2021-09-23 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2021-10-18 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Combined analysis:
| Analysis ID | AN003123 | AN003124 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | Reversed phase |
| Chromatography system | Thermo Dionex Ultimate 3000 RS | Thermo Dionex Ultimate 3000 RS |
| Column | Phenomenex Luna NH2 (150 x 2.1mm,3um) | Waters Acquity BEH C18 (150 x 2mm,1.7um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
| Ion Mode | UNSPECIFIED | POSITIVE |
| Units | counts | counts |
Chromatography:
| Chromatography ID: | CH002308 |
| Chromatography Summary: | For HILIC LC-MS/MS analysis, samples were analysed in positive and negative ion mode using a Phenomenex Luna NH2 analytical column (100 mm x 2 mm, 3 µm) held at room temperature with 10 minute linear gradient (A - 5% MeCN/20mM NH4CH2CO2/20mM NH4OH; B - MeCN) from 95 to 0 % MeCN followed by 5 minute hold at a flow rate of 0.400 mL/min. |
| Instrument Name: | Thermo Dionex Ultimate 3000 RS |
| Column Name: | Phenomenex Luna NH2 (150 x 2.1mm,3um) |
| Chromatography Type: | HILIC |
| Chromatography ID: | CH002309 |
| Chromatography Summary: | For reverse phase LC-MS/MS analysis, samples were analysed in positive ion mode only using a Waters Acquity BEH C18 analytical column (100 mm x 2.1 mm, 1.7 µm) held at 40oC with 4 minute linear gradient (A - H2O + 0.1% formic acid; B - MeOH + 0.1% formic acid) from 0.5 to 70% MeOH followed by a 5 minute hold at a flow rate of 0.350 mL/min. |
| Instrument Name: | Thermo Dionex Ultimate 3000 RS |
| Column Name: | Waters Acquity BEH C18 (150 x 2mm,1.7um) |
| Column Temperature: | 40 |
| Flow Gradient: | 4 minute linear gradient from 0.5 to 70% MeOH followed by a 5 minute hold |
| Flow Rate: | 0.350 mL/min |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% methanol; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
