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| Status |
Public on Nov 14, 2016 |
| Title |
The impact of low molecular weight hyaluronan on the lipidomic profile and global gene expression of peripheral blood mononuclear cells in asthma |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Hyaluronan (HA) is the major glycosaminoglycan in the extracellular matrix involved in the pathogenesis of asthma and other chronic inflammatory diseases. During inflammation there is an increased breakdown of HA, resulting in the local and systemic accumulation of low molecular weight (LMW) HA. Eicosanoids, derived from cytosolic phospholipase A2 group IVA activation, are potent lipid mediators also attributed to acute and chronic inflammation. We investigated the effect of LMW HA on lipidomic profile and global gene expression in PBMCs of patients with mild-to-moderate and severe asthma. We found that LMW HA increased production of 68 unique lipid species, among which PGE2, PGA2, PGD2, PGF2a, 15-HETE, TxB2, 11,12-EET, 14,15 EET, 13-HOTrE(y) and 16 (17) EpDPE were significantly upregulated in severe asthmatics. We also performed a systematic genome-wide expression analysis of LMW HA signaling, confirming its highly immunostimulatory potential. However, in severe asthmatics the LMW HA-induced global gene expression profile showed a comprehensive impairment in interferon signaling, cell apoptosis and cell movement, leading to diminished antiviral responses. Likewise, LMW HA-induced production of IL12 p40, CXCL10, CXCL11 and CCL8was markedly reduced in severe asthmatics. Our findings suggest a previously unforeseen link between extracellular matrix, global lipid production and antiviral responses in the pathogenesis of severe asthma.
We used microarray to perform a systematic genome-wide expression analysis of LMW HA effect in peripheral blood mononuclear cells in control subjects, mild-to moderate asthma and severe asthma patients
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| Overall design |
Blood specimens from patients with mild-to moderate asthma and severe asthma, and control subjects were drawn after obtaining informed consent. Blood samples were processed immediately using lymphocyte separation medium (LSM) (Lonza, Walkersville, MD), according to the manufacturer’s protocol to obtain peripheral blood monuclear cells (PBMCs). 4x106 PBMCs were resuspended in 1 ml of RPMI 1640 medium with 2 mM of L-glutamine, supplemented with 10% heat-inactivated FBS and treated with purified LMW HA (MP Biomedicals; Solon, OH) (stimulated samples) or vehicle (unstimulated samples) for 6 h. Each experiment was performed in duplicates. LMW HA used in this study was a purified mixture of fragments between 50 kDa to 600 kDa. Each experiment was performed in the presence of polymyxin B sulfate (Calbiochem) (10 ug/ml) to exclude an effect of LPS contamination. Supernatants were collected and kept frozen in -80oC before further analyses. Cell pellets were lysed in RLT buffer, homogenized using QIAshredder columns (Qiagen, Valencia, CA) and stored at -80C before total RNA extraction. Only one unstimulated and one stimulated sample was used for microarray per each study subject. There were 3 groups of subjects: control nonasthmatics (n=6), mild-to-moderate asthmatics (n=7), and severe asthmatics (n=6). In total we run 38 chips.
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| Contributor(s) |
Sokolowska M, Shelhamer JH |
| Citation(s) |
27826094 |
| Submission date |
Jul 02, 2014 |
| Last update date |
Dec 12, 2023 |
| Contact name |
Gabriela A Ferreyra |
| E-mail(s) |
gferreyra@cc.nih.gov
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| Phone |
301-435-2317
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| Organization name |
Clinical Center
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| Department |
CCMD
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| Street address |
9000 Rockville Pike - Building 10 room 4D03
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| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20892 |
| Country |
USA |
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| Platforms (1) |
| GPL15207 |
[PrimeView] Affymetrix Human Gene Expression Array |
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| Samples (38)
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| Relations |
| BioProject |
PRJNA254147 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE59019_RAW.tar |
73.2 Mb |
(http)(custom) |
TAR (of CEL) |
| Processed data included within Sample table |
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