Isolation of sensory hair cell specific exosomes in human perilymph

Zhuang, Pei; Phung, Suiching; Warnecke, Athanasia; Arambula, Alexandra; Peter, Madeleine St.; He, Mei; Staecker, Hinrich

doi:10.1016/j.neulet.2021.136282

Cited by 9 publications

(4 citation statements)

References 46 publications

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“…be EV associated. HCs are known to produce exosomes, which encase HC-specific proteins and can be detected in perilymph (30,31). Cochlear insult, such as with noise, is known to lead to increased prestin expression in the re-maining OHCs (32).…”

Section: Discussionmentioning

confidence: 99%

“…A closely related SLC26 gene family protein, Slc26a4—pendrin, has been shown to be present in exosomes from urine (29), indicating that structurally similar proteins can be EV associated. HCs are known to produce exosomes, which encase HC-specific proteins and can be detected in perilymph (30,31). Cochlear insult, such as with noise, is known to lead to increased prestin expression in the remaining OHCs (32).…”

Section: Discussionmentioning

confidence: 99%

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Automated Western Blot Analysis of Ototoxin-Induced Prestin Burst in the Blood after Cyclodextrin Exposure

Harrison,

Driscoll,

Farnsworth

et al. 2023

Otol Neurotol

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Hypothesis Ototoxin cyclodextrin (CDX) will induce a burst in serum prestin when quantified with automated Western blot analysis. Background In the clinical realm, we primarily rely on audiological measures for diagnosis and surveillance of sensorineural hearing loss (SNHL) and have limited therapeutic options. We have proposed a blood-based biomarker approach to overcome this challenge by measuring the outer hair cell’s (OHC) electromotile protein, prestin, in the blood. Previously, we demonstrated a burst in serum prestin after cisplatin exposure using enzyme-linked immunosorbent assayELISA. Methods Guinea pigs were treated with either 3,000 or 4,000 mg/kg CDX, and serum samples were obtained through 3 days after exposure. Serum prestin levels were quantified using automated blot analysis, western and hair cell counts were obtained. Results Both 3,000 and 4,000 mg/kg resulted in robust OHC loss, although more variability was seen at the lower dose. Automated Western blot analysis demonstrated that the prestin profile after CDX exposure is different than baseline. Specifically, a new ~134- kDa band accounted for the prestin burst after ototoxin ablation of OHCs at both doses. Conclusions We reproduced the prestin burst seen after cisplatin administration using CDX. Automated Western blot western analysis revealed that a ~a ~ 134- kDa species of prestin is responsible for the burst. We suggest that the induced band may be a prestin dimer, which could serve as a biomarker for early detection of ototoxicity in the clinical setting. These results add further promise to the potential of serum prestin to serve as an ototoxicity biomarker when using therapeutics with ototoxic properties.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Automated Western Blot Analysis of Ototoxin-Induced Prestin Burst in the Blood after Cyclodextrin Exposure

Harrison,

Driscoll,

Farnsworth

et al. 2023

Otol Neurotol

View full text Add to dashboard Cite

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“…The precise physiological role of exosomes is unclear, however they have been demonstrated to have pleiotropic roles in cell-cell communication, signaling, proliferation, angiogenesis, immune response and metabolism and have been shown to carry proteins, nucleic acids and metabolites ( Kalluri and LeBleu, 2020 ; Warnecke et al, 2022 ). Exosomes have been recently identified, isolated and characterized from the perilymph of adult patients with various forms of SNHL or mixed hearing loss, with the study suggesting a hair cell origin of the exosomes by their expression of MYO7A ( Zhuang et al, 2021 ). On the therapeutic side, a study from Athanasia Warnecke and colleagues demonstrated protective effects of bone marrow mesenchymal stromal cell-derived extracellular vesicles on a mouse model of NIHL, which additionally promoted neurite outgrowth in vitro ( Warnecke et al, 2020 ).…”

Section: Current and Emerging Therapeutic Optionsmentioning

confidence: 99%

Overcoming barriers: a review on innovations in drug delivery to the middle and inner ear

Delaney,

Liew,

Lye

et al. 2023

Front. Pharmacol.

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Despite significant advances in the development of therapeutics for hearing loss, drug delivery to the middle and inner ear remains a challenge. As conventional oral or intravascular administration are ineffective due to poor bioavailability and impermeability of the blood-labyrinth-barrier, localized delivery is becoming a preferable approach for certain drugs. Even then, localized delivery to the ear precludes continual drug delivery due to the invasive and potentially traumatic procedures required to access the middle and inner ear. To address this, the preclinical development of controlled release therapeutics and drug delivery devices have greatly advanced, with some now showing promise clinically. This review will discuss the existing challenges in drug development for treating the most prevalent and damaging hearing disorders, in particular otitis media, perforation of the tympanic membrane, cholesteatoma and sensorineural hearing loss. We will then address novel developments in drug delivery that address these including novel controlled release therapeutics such as hydrogel and nanotechnology and finally, novel device delivery approaches such as microfluidic systems and cochlear prosthesis-mediated delivery. The aim of this review is to investigate how drugs can reach the middle and inner ear more efficiently and how recent innovations could be applied in aiding drug delivery in certain pathologic contexts.

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“…In 2018, Wong et al (10) discovered the existence of exosomes in the inner ear and found that exosomes exert a protective effect against cisplatinand gentamicin-induced ototoxicity, thus suggesting their potential use as biomarkers. Breglio et al (11) found that exosomes also protect against aminoglycoside-induced hair cell death, and haircell-derived exosomes were found in the perilymph of patients with Meniere's disease, conductive/mixed hearing loss, and genetic SNHL (12). Furthermore, mesenchymal stromal/stem cell-derived exosomes alleviate cisplatin-induced ototoxicity (13)(14)(15).…”

Section: Introductionmentioning

confidence: 99%

Differentially expressed miRNA profiles of serum-derived exosomes in patients with sudden sensorineural hearing loss

Zhang

Yang

et al. 2023

Front. Neurol.

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ObjectivesThis study aimed to compare the expressed microRNA (miRNA) profiles of serum-derived exosomes of patients with sudden sensorineural hearing loss (SSNHL) and normal hearing controls to identify exosomal miRNAs that may be associated with SSNHL or serve as biomarkers for SSNHL.MethodsPeripheral venous blood of patients with SSNHL and healthy controls was collected to isolate exosomes. Nanoparticle tracking analysis, transmission electron microscopy, and Western blotting were used to identify the isolated exosomes, after which total RNA was extracted and used for miRNA transcriptome sequencing. Differentially expressed miRNAs (DE-miRNAs) were identified based on the thresholds of P < 0.05 and |log2fold change| > 1 and subjected to functional analyses. Finally, four exosomal DE-miRNAs, including PC-5p-38556_39, PC-5p-29163_54, PC-5p-31742_49, and hsa-miR-93-3p_R+1, were chosen for validation using quantitative real-time polymerase chain reaction (RT-qPCR).ResultsExosomes were isolated from serum and identified based on particle size, morphological examination, and expression of exosome-marker proteins. A total of 18 exosomal DE-miRNAs, including three upregulated and 15 downregulated miRNAs, were found in SSNHL cases. Gene ontology (GO) functional annotation analysis revealed that target genes in the top 20 terms were mainly related to “protein binding,” “metal ion binding,” “ATP binding,” and “intracellular signal transduction.” Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed that these target genes were functionally enriched in the “Ras,” “Hippo,” “cGMP-PKG,” and “AMPK signaling pathways.” The expression levels of PC-5p-38556_39 and PC-5p-29163_54 were significantly downregulated and that of miR-93-3p_R+1 was highly upregulated in SSNHL. Consequently, the consistency rate between sequencing and RT-qPCR was 75% and sequencing results were highly reliable.ConclusionThis study identified 18 exosomal DE-miRNAs, including PC-5p-38556_39, PC-5p-29163_54, and miR-93-3p, which may be closely related to SSNHL pathogenesis or serve as biomarkers for SSNHL.

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Isolation of sensory hair cell specific exosomes in human perilymph

Cited by 9 publications

References 46 publications

Automated Western Blot Analysis of Ototoxin-Induced Prestin Burst in the Blood after Cyclodextrin Exposure

Automated Western Blot Analysis of Ototoxin-Induced Prestin Burst in the Blood after Cyclodextrin Exposure

Overcoming barriers: a review on innovations in drug delivery to the middle and inner ear

Differentially expressed miRNA profiles of serum-derived exosomes in patients with sudden sensorineural hearing loss

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