Proteomic analysis of exosome-enriched fractions derived from cerebrospinal fluid of amyotrophic lateral sclerosis patients

Hayashi, Noriko; Doi, Hiroshi; Kurata, Yoichi; Kagawa, Hiroyuki; Atobe, Yoshitoshi; Funakoshi, Kengo; Tada, Mikiko; Katsumoto, Atsuko; Tanaka, Kenichi; Kunii, Misako; Nakamura, Hajime; Takahashi, Keita; Takeuchi, Hideyuki; Koyano, Shigeru; Kimura, Yayoi; Hirano, Hisashi; Tanaka, Fumiaki

doi:10.1016/j.neures.2019.10.010

Cited by 52 publications

(49 citation statements)

References 30 publications

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“…Though the findings of this study are corroborated by previously published data in in CSF exosomal mRNA and pathological data from post mortem tissue [25,27], further validation is required in larger cohorts to understand the clinical associations and robustness of these specific findings. One previous study of the CSF EV proteome in ALS did not detect these changes, though only 334 proteins were identified in that analysis and only univariate comparison of ALS and control proteomes was performed without exploring systematic differences within the entire proteome [14].…”

Section: Discussionmentioning

confidence: 99%

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CSF extracellular vesicle proteomics demonstrates altered protein homeostasis in amyotrophic lateral sclerosis

et al. 2020

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Background: Extracellular vesicles (EVs) released by neurons and glia reach the cerebrospinal fluid (CSF). Studying the proteome of CSF-derived EVs offers a novel perspective on the key intracellular processes associated with the pathogenesis of the neurodegenerative disease amyotrophic lateral sclerosis (ALS) and a potential source from which to develop biomarkers. Methods: CSF EVs were extracted using ultrafiltration liquid chromatography from ALS patients and controls. EV size distribution and concentration was measured using nanoparticle tracking analysis and liquid chromatographytandem mass spectrometry proteomic analysis performed. Results: CSF EV concentration and size distribution did not differ between ALS and control groups, nor between a subgroup of ALS patients with or without an associated hexanucleotide repeat expansion (HRE) in C9orf72. Univariate proteomic analysis identified downregulation of the pentameric proteasome-like protein Bleomycin hydrolase in ALS patients, whilst Gene Ontology enrichment analysis demonstrated downregulation of proteasome core complex proteins (8/8 proteins, normalized enrichment ratio-1.77, FDR-adjusted p = 0.057) in the ALS group. The subgroup of ALS patients associated with the C9orf72 HRE showed upregulation in Ubiquitin-like modifying-activating protein 1 (UBA1) compared to non-C9orf72 cases. Conclusions: Proteomic analysis of CSF EVs in ALS detects intracellular alterations in protein homeostatic mechanisms, previously only identified in pathological tissues. This supports the wider use of CSF EVs as a source of novel biomarkers reflecting key and potentially druggable pathological intracellular pathway alterations in ALS.

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Section: Discussionmentioning

confidence: 99%

“…studies have yet taken this approach [14]. This study used an optimised high-yield, high-purity CSF EV extraction method combined with quantification and subsequent proteomic analysis of EVs extracted from CSF of patients with ALS and controls to identify novel candidate biomarkers of ALS.…”

Section: Introductionmentioning

confidence: 99%

CSF extracellular vesicle proteomics demonstrates altered protein homeostasis in amyotrophic lateral sclerosis

et al. 2020

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“…Consistent with the role of inflammation in ALS, Chen and colleagues have found elevated levels of interleukin-6 (IL-6) in astrocyte-derived exosomes retrieved from plasma of sALS patients, in addition to a positive correlation between their levels and rate of disease progression [84]. A proteomic study performed on exosome-enriched fraction of CSF has revealed increased concentration of novel INHAT repressor (NIR), involved in nucleolar function, in ALS patients [85]; interestingly NIR has shown altered subcellular localization in MNs of affected patients.…”

Section: Evs As Biomarkers Of Diagnosis and Prognosismentioning

confidence: 84%

Extracellular vesicles and amyotrophic lateral sclerosis: from misfolded protein vehicles to promising clinical biomarkers

Gagliardi

Bresolin

Comi

et al. 2020

Cell. Mol. Life Sci.

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Extracellular vesicles (EVs) are small reservoirs of different molecules and important mediators of cell-to-cell communication. As putative vehicles of misfolded protein propagation between cells, they have drawn substantial attention in the field of amyotrophic lateral sclerosis (ALS) and other neurodegenerative disorders. Moreover, exosome-mediated non-coding RNA delivery may play a crucial role in ALS, given the relevance of RNA homeostasis in disease pathogenesis. Since EVs can enter the systemic circulation and are easily detectable in patients' biological fluids, they have generated broad interest both as diagnostic and prognostic biomarkers and as valuable tools in understanding disease pathogenesis. Here, after a brief introduction on biogenesis and functions of EVs, we aim to investigate their role in neurodegenerative disorders, especially ALS. Specifically, we focus on the main findings supporting EV-mediated protein and RNA transmission in ALS in vitro and in vivo models. Then, we provide an overview of clinical applications of EVs, summarizing the most relevant studies able to detect EVs in blood and cerebrospinal fluid (CSF) of ALS patients, underlying their potential use in aiding diagnosis and prognosis. Finally, we explore the therapeutic applications of EVs in ALS, either as targets or as vehicles of proteins, nucleic acids and molecular drugs.

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“…To facilitate the clinical application of exosomes, efficient separation and storage methods are essential. The current methods for isolating exosomes primarily comprise ultracentrifugation (Baranyai et al, 2015), the microfluidic chip method (Zhang et al, 2019), the antibody affinity capture method (Mathivanan et al, 2010), the polymer precipitation method (Cao F. et al, 2019), and gel exclusion chromatography (Hayashi et al, 2019). Of these, the most commonly used method is ultracentrifugation.…”

Section: Exosome Isolation and Storagementioning

confidence: 99%

Circulating Exosomal miRNA as Diagnostic Biomarkers of Neurodegenerative Diseases

Wang

Zhang

2020

Front. Mol. Neurosci.

115

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Neurodegenerative diseases (NDDs) are a group of diseases caused by chronic and progressive degeneration of neural tissue. The main pathological manifestations are neuronal degeneration and loss in the brain and/or spinal cord. Common NDDs include Alzheimer disease (AD), Parkinson disease (PD), Huntington disease (HD), and amyotrophic lateral sclerosis (ALS). The complicated pathological characteristics and different clinical manifestations of NDDs result in a lack of sensitive and efficient diagnostic methods. In addition, no sensitive biomarkers are available to monitor the course of NDDs, predict their prognosis, and monitor the therapeutic response. Despite extensive research in recent years, analysis of amyloid β (Aβ) and α-synuclein has failed to effectively improve NDD diagnosis. Although recent studies have indicated circulating miRNAs as promising diagnostic biomarkers of NDDs, the miRNA in the peripheral circulation is susceptible to interference by other components, making circulating miRNA results less consistent. Exosomes are small membrane vesicles with a diameter of approximately 30-100 nm that transport proteins, lipids, mRNA, and miRNA. Because recent studies have shown that exosomes have a double-membrane structure that can resist ribonuclease in the blood, giving exosomal miRNA high stability and making them resistant to degradation, they may become an ideal biomarker of circulating fluids. In this review, we discuss the applicability of circulating exosomal miRNAs as biomarkers, highlight the technical aspects of exosomal miRNA analysis, and review studies that have used circulating exosomal miRNAs as candidate diagnostic biomarkers of NDDs.

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Proteomic analysis of exosome-enriched fractions derived from cerebrospinal fluid of amyotrophic lateral sclerosis patients

Cited by 52 publications

References 30 publications

CSF extracellular vesicle proteomics demonstrates altered protein homeostasis in amyotrophic lateral sclerosis

CSF extracellular vesicle proteomics demonstrates altered protein homeostasis in amyotrophic lateral sclerosis

Extracellular vesicles and amyotrophic lateral sclerosis: from misfolded protein vehicles to promising clinical biomarkers

Circulating Exosomal miRNA as Diagnostic Biomarkers of Neurodegenerative Diseases

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