PXD065520

PXD065520 is an original dataset announced via ProteomeXchange.

Title	Quantitative Proteomics Reveals Fh15 as an Antagonist of TLR4 Downregulating the Activation of NF-κB, Inducible Nitric Oxide, Phagosome Signaling Pathways, and Oxidative Stress of LPS-stimulated Macrophages
Description	There is a present need to develop alternative biotherapeutic drugs to mitigate the exacerbated inflammatory immune responses characteristic of sepsis. The potent endotoxin lipopolysaccharide (LPS), a major component of Gram-negative bacterial outer membrane, activates the immune system via Toll-like receptor 4 (TLR4), triggering macrophages and a persistent cascade of inflammatory mediators. Our previous studies have demonstrated that Fh15, a recombinant member of the Fasciola hepatica fatty acid binding protein family, can significantly increase the survival rate by suppressing many inflammatory mediators induced by LPS in a septic shock mouse model. Although Fh15 has been proposed as a TLR4 antagonist, the specific mechanisms underlying its immune modulatory effect remained unclear. In the present study we employed a quantitative proteomics approach using tandem mass tag (TMT) followed by LC-MS/MS analysis to identify and quantify differentially expressed proteins that participate in signaling pathways downstream TLR4 of macrophages, which can be dysregulated by Fh15. Based on significant fold change (FC) cutoff of 1.5 and p-value ≤ 0.05 criteria, we focused our attention to 114 proteins that were upregulated by LPS and downregulated by Fh15. From these proteins, TNFα, IL-1𝛼, Lck, NOS2, SOD2 and CD36 were selected for validation by Western blot on murine bone marrow derived macrophages due to their relevant roles in the NF-κB, iNOS, oxidative stress, and phagosome signaling pathways, which are closely associated to sepsis pathogenesis. These results suggest that Fh15 exerts a broad spectrum of action by simultaneously targeting multiple downstream pathways activated by TLR4, thereby modulating various aspects of the inflammatory responses during sepsis.
HostingRepository	PRIDE
AnnounceDate	2025-08-04
AnnouncementXML	Submission_2025-08-03_16:33:08.259.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD065520
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Loyda Melendez
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	TMT6plex-126 reporter+balance reagent acylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2025-06-26 09:19:25	ID requested
⏵ 1	2025-08-03 16:33:09	announced

Armina-Rodriguez A, Vald, é, s Fernandez BN, Ocasio-Malav, é C, Cantres Rosario YM, Carrasquillo Carri, ó, n K, Mel, é, ndez LM, Roche Lima A, Tosado Rodriguez EL, Espino AM, B, Inducible Nitric Oxide, Phagosome Signaling Pathways, and Oxidative Stress of LPS-Stimulated Macrophages. Int J Mol Sci, 26(14):(2025) [pubmed]

10.6019/PXD065520;

10.3390/ijms26146914;

submitter keyword: TMT-labeling

Proteomics

Fasciola hepatica

Fh15

TLR

NF-κB

iNOS

CD36

Lck

LPS

macrophages

sepsis

Loyda M Melendez
contact affiliation	Translational Proteomics Center, University of Puerto RIco, Medical Sciences Campus
contact email	lmelendezlab@gmail.com
lab head
Loyda Melendez
contact affiliation	University of Puerto Rico Medical Sciences Campus
contact email	lmelendezlab@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD065520
     1. Label: PRIDE project
     2. Name: Quantitative Proteomics Reveals Fh15 as an Antagonist of TLR4 Downregulating the Activation of NF-κB, Inducible Nitric Oxide, Phagosome Signaling Pathways, and Oxidative Stress of LPS-stimulated Macrophages