Proteomic Analysis of Two Types of Exosomes in Human Whole Saliva

Ogawa, Yuko; Miura, Yuri; Harazono, Akira; Kanai‐Azuma, Masami; Akimoto, Yoshihiro; Kawakami, Hayato; Yamaguchi, Teruhide; Toda, Tosifusa; Endo, Tamao; Tsubuki, Masayoshi; Yanoshita, Ryohei

doi:10.1248/bpb.34.13

Cited by 220 publications

(184 citation statements)

References 47 publications

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“…For example, EVs offer high physicochemical and biological stability,11 which may be due to the isolating ability of the phospholipid layer, protecting the contents of EVs from exposure to the external environment ( Figure 2 ). 12 As a result, EVs can exist in almost all body fluids, including blood plasma,13 saliva,14 urine,15 bile,16 synovial fluid,17 semen,18 and breast milk 19. By expressing different surface proteins, EVs can have different targeting abilities 20.…”

Section: Introductionmentioning

confidence: 99%

Modularized Extracellular Vesicles: The Dawn of Prospective Personalized and Precision Medicine

2018

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Extracellular vesicles (EVs) are ubiquitous nanosized membrane vesicles consisting of a lipid bilayer enclosing proteins and nucleic acids, which are active in intercellular communications. EVs are increasingly seen as a vital component of many biological functions that were once considered to require the direct participation of stem cells. Consequently, transplantation of EVs is gradually becoming considered an alternative to stem cell transplantation due to their significant advantages, including their relatively low probability of neoplastic transformation and abnormal differentiation. However, as research has progressed, it is realized that EVs derived from native‐source cells may have various shortcomings, which can be corrected by modification and optimization. To date, attempts are made to modify or improve almost all the components of EVs, including the lipid bilayer, proteins, and nucleic acids, launching a new era of modularized EV therapy through the “modular design” of EV components. One high‐yield technique, generating EV mimetic nanovesicles, will help to make industrial production of modularized EVs a reality. These modularized EVs have highly customized “modular design” components related to biological function and targeted delivery and are proposed as a promising approach to achieve personalized and precision medicine.

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Section: Introductionmentioning

confidence: 99%

Modularized Extracellular Vesicles: The Dawn of Prospective Personalized and Precision Medicine

2018

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confidence: 99%

“…13,14) Although our method has not been universally recognized for purifying exosomes, Wyss et al recently showed that ultrafiltration and size-exclusion are valid methodologies for intact exosome purification. 15) Because exosomal marker proteins (Alix, tsg101, hsp70 and CD63) were detected in both samples by Western blot analysis, we designated the vesicles as exosomes I and II.…”

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confidence: 99%

Next-Generation Sequencing of Protein-Coding and Long Non-protein-Coding RNAs in Two Types of Exosomes Derived from Human Whole Saliva

Ogawa

Tsujimoto

Yanoshita

2016

Biological & Pharmaceutical Bulletin

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Exosomes are small extracellular vesicles containing microRNAs and mRNAs that are produced by various types of cells. We previously used ultrafiltration and size-exclusion chromatography to isolate two types of human salivary exosomes (exosomes I, II) that are different in size and proteomes. We showed that salivary exosomes contain large repertoires of small RNAs. However, precise information regarding long RNAs in salivary exosomes has not been fully determined. In this study, we investigated the compositions of protein-coding RNAs (pcRNAs) and long non-protein-coding RNAs (lncRNAs) of exosome I, exosome II and whole saliva (WS) by next-generation sequencing technology. Although 11% of all RNAs were commonly detected among the three samples, the compositions of reads mapping to known RNAs were similar. The most abundant pcRNA is ribosomal RNA protein, and pcRNAs of some salivary proteins such as S100 calcium-binding protein A8 (protein S100-A8) were present in salivary exosomes. Interestingly, lncRNAs of pseudogenes (presumably, processed pseudogenes) were abundant in exosome I, exosome II and WS. Translationally controlled tumor protein gene, which plays an important role in cell proliferation, cell death and immune responses, was highly expressed as pcRNA and pseudogenes in salivary exosomes. Our results show that salivary exosomes contain various types of RNAs such as pseudogenes and small RNAs, and may mediate intercellular communication by transferring these RNAs to target cells as gene expression regulators.Key words exosome; transcriptome; human whole saliva; next-generation sequencing; non-coding RNA; pseudogene Human whole saliva (WS) contains an aqueous complex mixture of proteins, peptides, hormones, metabolites, DNAs and RNAs. WS contributes to maintain the integrity of the oral cavity through its lubricating, antibacterial, antiviral and buffering actions, and facilitates chewing and swallowing food. It plays an important role in front-line body defense. Because WS can be collected simply, cheaply and noninvasively, it has been used to monitor human health and disease. Because saliva contains ribonucleases and nucleases from various sources, which influence RNA stability, relatively few studies have analyzed human salivary RNA.1) Salivary nucleotides are protected from degradation by inclusion in extracellular vesicles such as exosomes. Recently, RNAs and DNAs in WS have been highlighted in the fields of biomarker research, disease diagnostics and forensic study. 2-4)Exosomes are small (30-100 nm) membrane vesicles of endocytic origin that are released into the extracellular environment upon fusion of multivesicular bodies with the plasma membrane. Exosomes are present in various body fluids including blood, breast milk, malignant ascites, urine, amniotic fluid and saliva. 5) Exosomes can contain the proteins and nucleic acids of their cell of origin and can transfer their contents to recipient cells at a distance. 5) Exosomes are now thought to be secreted by various cell types, and numerous comp...

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“…Although eHSP70 derived from periodontal tissue may co-mix into the saliva samples via GCF, no subject had orthodontic appliances in this study. In addition, Lancaster and Febbraio (2005) showed that exosomes contributed to circulating eHSP70 levels, and Ogawa et al (2011) reported that exosomes exists in saliva. Therefore, most eHSP70 in the collected saliva samples was likely derived from either the salivary glands, oral mucosa, or salivary exosomes.…”

Section: Discussionmentioning

confidence: 99%

Salivary extracellular heat shock protein 70 (eHSP70) levels increase after 59 min of intense exercise and correlate with resting salivary secretory immunoglobulin A (SIgA) levels at rest

Murase

Shimizu

Tanimura

et al. 2016

Cell Stress and Chaperones

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This study aimed to identify the response of a salivary stress protein, extracellular heat shock protein (eHSP70), to intense exercise and to investigate the relationship between salivary eHSP70 and salivary immunoglobulin A (SIgA) levels in response to exercise. Sixteen healthy sedentary young males (means±SD 23.8±1.5 years, 172.2±6.4 cm, 68.3±7.4 kg) performed 59 min of cycling exercise at 75 % VO2 max . Saliva and whole blood samples were collected before (Pre), immediately after (Post), and at 1, 2, 3, and 4 h after completion of the exercise (1, 2, 3, and 4 h). The salivary eHSP70 and SIgA levels were measured by enzyme-linked imunosorbent assay (ELISA), and the secretion rates were computed by multiplying the concentration by the saliva flow rate. White blood cells were analyzed using an automated cell counter with a directcurrent detection system. The salivary eHSP70 secretion rates were 1

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Proteomic Analysis of Two Types of Exosomes in Human Whole Saliva

Cited by 220 publications

References 47 publications

Modularized Extracellular Vesicles: The Dawn of Prospective Personalized and Precision Medicine

Modularized Extracellular Vesicles: The Dawn of Prospective Personalized and Precision Medicine

Next-Generation Sequencing of Protein-Coding and Long Non-protein-Coding RNAs in Two Types of Exosomes Derived from Human Whole Saliva

Salivary extracellular heat shock protein 70 (eHSP70) levels increase after 59 min of intense exercise and correlate with resting salivary secretory immunoglobulin A (SIgA) levels at rest

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