Dataset Information

Compartmentalisation proteomics revealed endolysosomal protein network changes in a goat model of atrial fibrillation

ABSTRACT: The emerging roles of endolysosomes (EL) in AF have yet to be discovered, especially how the EL contribute to the development of AF within a slow time frame. To identify the EL role in AF, we applied proteomics, transcriptomics, integrated analysis, electron tomography, western blotting and enzyme assays. To understand the cellular and EL driven dominant pathways, in AF, we applied an organelle protein isolation method, that was previously developed by us (Ayagama et al,.2021) on to an AF goat model. Our findings include upregulation of AMPK pathway, EL-specific proteins that were not identified in the TL, such as GAA, DYNLRB1, CLTB, SIRT3, CCT2, and muscle specific HSPB2. The structural anomalies such as autophago-vacuole formation, irregularly stacked mitochondria, and glycogen deposition provide an insight into of the contribution of EL contribution and the related pathways and molecular mechanisms.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Capra Hircus X Ovis Aries

TISSUE(S): Heart

SUBMITTER: Georgina Berridge

LAB HEAD: Rebecca AB Burton

PROVIDER: PXD041056 | Pride | 2024-06-16

REPOSITORIES: Pride

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Compartmentalization proteomics revealed endolysosomal protein network changes in a goat model of atrial fibrillation.

Ayagama Thamali T Charles Philip D PD Bose Samuel J SJ Boland Barry B Priestman David A DA Aston Daniel D Berridge Georgina G Fischer Roman R Cribbs Adam P AP Song Qianqian Q Mirams Gary R GR Amponsah Kwabena K Heather Lisa L Galione Antony A Herring Neil N Kramer Holger H Capel Rebecca A RA Platt Frances M FM Schotten Ulrich U Verheule Sander S Burton Rebecca A B RAB

iScience 20240328 6

Endolysosomes (EL) are known for their role in regulating both intracellular trafficking and proteostasis. EL facilitate the elimination of damaged membranes, protein aggregates, membranous organelles and play an important role in calcium signaling. The specific role of EL in cardiac atrial fibrillation (AF) is not well understood. We isolated atrial EL organelles from AF goat biopsies and conducted a comprehensive integrated omics analysis to study the EL-specific proteins and pathways. We also ...[more]

PMID: 38827406

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Project description:Single-walled carbon nanotubes (SWCNTs) are being explored and used for a wide range of applications in industrial and medical sectors, and the increasing exposure of SWCNTs necessitate the studies of their potential environmental and health effects. Considerable efforts have been made to improve the dispersion of SWCNTs by chemical modifications. The present study was designed to determine if the acid functionalization of SWCNTs enhanced the toxicity and more efforts were made to understand the molecular mechanisms. RAW264.7 cells were exposed to 0-100μg/mL of SWCNTs and AF-SWCNTs for 24 hours and cell proliferation, viability, and gene expression profiles were assessed and compared. Results showed that the magnitude of AF-SWCNT-induced cell proliferation inhibition was higher than that of SWCNTs. 10μg/mL of AF-SWCNTs was determined as non-toxic to cells by viability analysis. AF-SWCNTs could enter and aggregate in cell cytoplasm and nuclear areas. Microarray, real-time reverse-transcriptase polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assays (ELISA) demonstrated that AF-SWCNTs (1, 10, and 50μg/mL) altered gene and protein expression patterns. The most differentially expressed genes were related to pro-inflammatory cytokines and chemokines (such as CCL-3 and CCL-4), apoptosis (e.g., Caspase-1, -4, and Dffa), protective response (such as Hmox-1and GSTM3), and cell surface molecules such as ICAM-1, Itgb2, indicating a possible involvement of SWCNT in inflammation, anti-apoptotic and DNA fragmentation, carcinogenesis and biased T cell polarization. Furthermore, a substantial amount of down-regulated genes were found related to ribosomal assembly and mitochondrial respiration chain, implicating a possible mechanism of SWCNT-induced oxidative stress and gene expression inhibition. RAW264.7 gene expression after exposure to 1-50ug/ml AF-SWCNT-24 hours. 0ug/ml of AF-SWCNT was used as control sample. The data were obtained from three biological replicates.

Project description:Atrial fibrillation (AF) is the most common sustained arrhythmia characterized by rapid and multiple irregular excitations within the atria. AF is associated with serious morbidity and increased mortality, and its prevalence is prospected to increase as society ages. The limited therapeutic efficacy of AF treatment as well as its high socioeconomic burden makes AF a major clinical challenge. Despite our expanding knowledge of individual proteins and pathways involved in the complex pathophysiology of atrial fibrillation (AF), an unbiased overview of proteins and functionally enriched biological processes as well as their crosstalk is lacking. Here, we performed an explorative proteomics analysis to reveal the global abundance of proteins in cardiac tissue of patients, and deciphered functionally grouped gene ontologies (GO) to uncover a perspective of the disease biology driving or driven by AF. A total of 2703 proteins were identified by liquid chromatography coupled to tandem mass spectrometry. Among them, 150 proteins (accounting for 5.6% of 2703) had a significantly altered abundance (100 proteins increased and 50 decreased) in AF. A significant biological connection was found between those (protein-protein interaction enrichment p-value=1.0e-16). GO enrichment analysis showed that these 150 proteins were mainly located in extracellular/cytoplasmic vesicles, mitochondrion, and cytoskeletal compartments. Correspondingly, the 100 proteins increased in AF were significantly enriched in the GO terms related to immune system, metabolic process, iron process, ECM disassembly, mitochondrial translation and apoptotic signaling. Partially clustered proteins with dense functional link were found in immune system and metabolic process, and were respectively annotated in neutrophil degranulation, and oxoacid metabolic process coupled to the subunits of mitochondrial dehydrogenase NADH. Those processes enriched in AF had crosstalk via the proteins involved in neutrophil degranulation. Selected proteins such as LCN2 (neutrophil degranulation), CA3 (immune system), NDUFS2 (complex I) and MYH10 (actin motor protein) were validated by western blot or qPCR in an independent cohort. The 50 proteins decreased in AF were collectively enriched in vesicle-mediated transport and actin filament-based movement. We demonstrate that important biological processes underlying persistent AF as well as their crosstalk via the components of neutrophil degranulation. Our study provides a novel insight for a more efficient targeting strategy for AF treatment.

Dataset Information

Compartmentalisation proteomics revealed endolysosomal protein network changes in a goat model of atrial fibrillation

Publications

Compartmentalization proteomics revealed endolysosomal protein network changes in a goat model of atrial fibrillation.

Similar Datasets

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