Descriptive proteomics of paired human vocal fold and buccal mucosa tissue

Grossmann, Tanja; Darnhofer, Barbara; Birner‐Gruenberger, Ruth; Kirsch, Andrijana; Gugatschka, Markus

doi:10.1002/prca.202100050

Cited by 3 publications

(4 citation statements)

References 23 publications

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“…These proteins include structural components of the extracellular matrix such as collagens, fibrillin 1 and 2, fibulins, elastins, laminins, proteoglycans, decorin and fibronectin; and muscle proteins such as myosins and actins. These proteins are also present in human vocal folds [ 57 ], corroborating our protein extraction and fractionation methods and the LC-MS/MS approach used to analyze the proteome of rabbit vocal folds.…”

Section: Discussionsupporting

confidence: 65%

“…These results indicate that the vocal folds have independent mechanisms of protein regulation related to the aging process, response to hydration status, and a combination of age and level of hydration (interaction). Despite the identification of unique proteins regulated by the individual effects considered in this study, the enrichment pathway analysis showed a common association of these proteins to extracellular matrix organization and muscle structure/contraction, suggesting that age and hydration, as single or interacting factors, interfere with the physiological biomechanical and viscoelastic properties of the vocal folds responsible for structural support and voice production [ 57 – 62 ].…”

Section: Discussionmentioning

confidence: 99%

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Proteomic analysis reveals that aging rabbit vocal folds are more vulnerable to changes caused by systemic dehydration

Nascimento¹,

Bailey²,

Santos³

et al. 2022

BMC Genomics

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Background Older adults are more prone to develop systemic dehydration. Systemic dehydration has implications for vocal fold biology by affecting gene and protein expression. The objective of this study was to quantify vocal fold protein changes between two age groups and hydration status, and to investigate the interaction of age and hydration status on protein expression, which has not been investigated in the context of vocal folds before. Comparative proteomics was used to analyze the vocal fold proteome of 6.5-month-old and > 3-year-old rabbits subjected to water ad libitum or water volume restriction protocol. Results Young and older adult rabbits (n = 22) were either euhydrated (water ad libitum) or dehydrated by water volume restriction. Dehydration was confirmed by body weight loss of − 5.4% and − 4.6% in young and older groups, respectively, and a 1.7-fold increase of kidney renin gene expression in the young rabbits. LC-MS/MS identified 2286 proteins in the rabbit vocal folds of young and older adult rabbits combined. Of these, 177, 169, and 81 proteins were significantly (p ≤ 0.05) affected by age, hydration status, or the interaction of both factors, respectively. Analysis of the interaction effect revealed 32 proteins with opposite change patterns after dehydration between older and young rabbit vocal folds, while 31 proteins were differentially regulated only in the older adult rabbits and ten only in the young rabbits in response to systemic dehydration. The magnitude of changes for either up or downregulated proteins was higher in the older rabbits. These proteins are predominantly related to structural components of the extracellular matrix and muscle layer, suggesting a disturbance in the viscoelastic properties of aging vocal fold tissue, especially when subjected to systemic dehydration. Conclusions Water restriction is a laboratory protocol to assess systemic dehydration-related changes in the vocal fold tissue that is translatable to human subjects. Our findings showed a higher number of proteins differentially regulated with a greater magnitude of change in the vocal folds of older adult rabbits in the presence of systemic dehydration compared to younger rabbits. The association of these proteins with vocal fold structure and biomechanical properties suggests that older human subjects may be more vulnerable to the effects of systemic dehydration on vocal function. The clinical implications of these protein changes warrant more investigation, but age should be taken into consideration when evaluating vocal treatment recommendations that interfere with body fluid balance.

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Section: Discussionsupporting

confidence: 65%

Section: Discussionmentioning

confidence: 99%

Proteomic analysis reveals that aging rabbit vocal folds are more vulnerable to changes caused by systemic dehydration

Nascimento¹,

Bailey²,

Santos³

et al. 2022

BMC Genomics

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“…Since BM epithelial cells are more easily available compared to hVFE, we decided to include this cell type in a 3D co-culture setting with hVFF. This approach was supported by the results of a previous study from our group, which showed a high similarity between mucosal VF and BM tissue at the proteomic level [4]. The VF epithelium is the first cellular layer and consists of five to ten closely packed non-keratinized stratified squamous cell layers, in contrast to keratinized cell layers, such as those found in the skin [18].…”

Section: Plos Onesupporting

confidence: 61%

“…the mucosa can be harvested during arytenoidectomy or from postmortem biopsies), human VF epithelium which is required for these experiments, is only available in exceedingly small numbers [3], so alternatives for this cell type are urgently needed. Epithelial cells of buccal mucosal origin could be such a candidate with similar properties, as they are easy to harvest, available in sufficient numbers, and biopsies heal without scarring [4].…”

Section: Introductionmentioning

confidence: 99%

Introducing a new type of alternative laryngeal mucosa model

et al. 2023

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Research of human vocal fold (VF) biology is hampered by several factors. The sensitive microstructure of the VF mucosa is one of them and limits the in vivo research, as biopsies carry a very high risk of scarring. A laryngeal organotypic model consisting of VF epithelial cells and VF fibroblasts (VFF) may overcome some of these limitations. In contrast to human VFF, which are available in several forms, availability of VF epithelial cells is scarce. Buccal mucosa might be a good alternative source for epithelial cells, as it is easily accessible, and biopsies heal without scarring. For this project, we thus generated alternative constructs consisting of immortalized human VF fibroblasts and primary human buccal epithelial cells. The constructs (n = 3) were compared to native laryngeal mucosa at the histological and proteomic level. The engineered constructs reassembled into a mucosa-like structure after a cultivation period of 35 days. Immunohistochemical staining confirmed a multi-layered stratified epithelium, a collagen type IV positive barrier-like structure resembling the basement membrane, and an underlying layer containing VFF. Proteomic analysis resulted in a total number of 1961 identified and quantified proteins. Of these, 83.8% were detected in both native VF and constructs, with only 53 proteins having significantly different abundance. 15.3% of detected proteins were identified in native VF mucosa only, most likely due to endothelial, immune and muscle cells within the VF samples, while 0.9% were found only in the constructs. Based on easily available cell sources, we demonstrate that our laryngeal mucosa model shares many characteristics with native VF mucosa. It provides an alternative and reproducible in vitro model and offers many research opportunities ranging from the study of VF biology to the testing of interventions (e.g. drug testing).

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Descriptive proteomics of paired human vocal fold and buccal mucosa tissue

Grossmann

Darnhofer

Birner‐Gruenberger

et al. 2021

Proteomics Clinical Apps

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The vast majority of voice disorders is associated with changes of the unique, but delicate, human vocal fold mucosa. The ability to develop new effective treatment methods is significantly limited by the physical inaccessibility and the extremely rare occasions under which healthy tissue biopsies can be obtained. Therefore, the interest in laryngological research has shifted to human oral (buccal) mucosa, a similar and more easily available tissue. The harvesting process is less invasive and accompanied with faster healing and less scarring, compared to vocal fold mucosa. Here we report a descriptive proteomic comparison of paired human buccal and vocal fold mucosa by high‐resolution mass spectrometry (CID‐MS/MS). Our study identified a total of 1575 proteins detected within both tissues that are highly consistent in several crucial biological processes, cellular components, and molecular functions. Hence, our proteomic analysis will provide a fundamental resource for the laryngological research community.

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Descriptive proteomics of paired human vocal fold and buccal mucosa tissue

Cited by 3 publications

References 23 publications

Proteomic analysis reveals that aging rabbit vocal folds are more vulnerable to changes caused by systemic dehydration

Proteomic analysis reveals that aging rabbit vocal folds are more vulnerable to changes caused by systemic dehydration

Introducing a new type of alternative laryngeal mucosa model

Descriptive proteomics of paired human vocal fold and buccal mucosa tissue

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