Mycoplasma affects baseline gene expression and the response to glucocorticoids in vocal fold fibroblasts

Carina Doyle; Ryosuke Nakamura; Renjie Bing; Bernard Rousseau; Ryan C. Branski

doi:10.1099/jmm.0.001362

Volume 70, Issue 5

Short Communication

Open Access

Mycoplasma affects baseline gene expression and the response to glucocorticoids in vocal fold fibroblasts

Carina Doyle¹, Ryosuke Nakamura¹, Renjie Bing¹, Bernard Rousseau² and Ryan C. Branski^1,3
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Affiliations: ¹ Department of Rehabilitation Medicine, NYU Grossman School of Medicine, New York, NY, USA ² Communication Sciences and Disorders, School of Health and Rehabilitation Sciences, University of Pittsburgh, Pittsburgh, PA, USA ³ Department of Otolaryngology-Head and Neck Surgery, NYU Grossman School of Medicine, New York, NY, USA
*Correspondence: Ryan C. Branski, [email protected]
Published: 26 May 2021 https://doi.org/10.1099/jmm.0.001362

Abstract

Introduction. In vitro experimentation is intentionally contrived to isolate specific phenomena in the context of profound biological complexity. Mycoplasmas in the upper airway likely contribute to this complexity and play a largely unknown role in both health and disease. Similarly, the presence and role of mycoplasma in in vitro investigation are largely unknown.

Hypothesis. We hypothesize mycoplasma in human vocal fold fibroblasts (VFF) will affect both basal gene-expression patterns as well as the cell response to exogenous stimuli.

Aim. We sought to determine mycoplasma presence across vocal fold fibroblast cultures, basal transcriptional changes as a function of mycoplasma, and responsiveness to exogenous glucocorticoids in mycoplasma-positive and -negative VFF.

Methodology. PCR-based mycoplasma detection was performed in an immortalized human VFF line as well as rat and rabbit primary VFF cultures and extracted rat laryngeal tissue. RNA sequencing was performed in mycoplasma-positive and -negative human cells at baseline and in response to dexamethasone.

Results. Mycoplasma was identified in the human cell line as well as primary culture from rabbits. Mycoplasma was not detected in tissue or primary culture from rat vocal folds. Basal mRNA expression in human VFF differed significantly following mycoplasma treatment. In addition, differential responses to dexamethasone were observed across multiple pathways as a function of mycoplasma presence in these cells. Pathways including apoptosis, DNA damage repair, and G1 to S cell cycle signalling were significantly enriched in mycoplasma-positive cells.

Conclusion. Variability of mycoplasma presence across culture conditions and differential responses to exogenous stimuli as a function of mycoplasma presence are potentially problematic for the translation of in vitro experimentation in the upper aerodigestive tract. It remains unclear if these findings represent contamination or the baseline state of this specialized tissue.

Received: 10/12/2020
Accepted: 08/04/2021
Published Online: 26/05/2021

Keyword(s): dexamethasone , fibroblast , mycoplasma , steroids , vocal fold and voice

Funding

This study was supported by the:

National Institutes of Health/National Institute on Deafness and Other Communication Disorders (Award RO1 DC017397)
- Principle Award Recipient: RyanBranski

This is an open-access article distributed under the terms of the Creative Commons Attribution NonCommercial License.

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Mycoplasma affects baseline gene expression and the response to glucocorticoids in vocal fold fibroblasts

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Volume 70, Issue 5

Short Communication

Open Access

Mycoplasma affects baseline gene expression and the response to glucocorticoids in vocal fold fibroblasts

Abstract

Funding

Supplementary material 1

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