Mycoplasma affects baseline gene expression and the response to glucocorticoids in vocal fold fibroblasts.
Mycoplasma affects baseline gene expression and the response to glucocorticoids in vocal fold fibroblasts.
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DOI：
10.1099/jmm.0.001362
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发表时间：
2021-05
影响因子：
3
通讯作者：
Branski RC
中科院分区：
文献类型：
--
作者：
Doyle C;Nakamura R;Bing R;Rousseau B;Branski RC
体外实验是有意设计的，以隔离在深刻的生物复杂性的背景下的特定现象。上呼吸道中的支原体可能导致这种复杂性，并在健康和疾病中发挥很大程度上未知的作用。同样，支原体在体外研究中的存在和作用在很大程度上是未知的。我们假设人声带成纤维细胞（VFF）中的支原体会影响基础基因表达模式以及细胞对外源性刺激的反应。我们试图确定支原体存在声带成纤维细胞培养，作为支原体的功能，基础的转录变化，并在支原体阳性和阴性VFF外源性糖皮质激素的反应。在永生化人VFF系以及大鼠和兔原代VFF培养物和提取的大鼠喉组织中进行基于PCR的支原体检测。在基线和地塞米松治疗后，对支原体阳性和阴性人细胞进行RNA测序。在人细胞系以及兔原代培养物中鉴别出支原体。在大鼠声带组织或原代培养物中未检出支原体。支原体处理后，人VFF中的基础mRNA表达显着不同。此外，作为这些细胞中支原体存在的函数，在多个途径中观察到对地塞米松的不同反应。包括细胞凋亡，DNA损伤修复和G1到S细胞周期信号转导的途径显着丰富的支原体阳性细胞。在培养条件下支原体存在的变异性和对外源性刺激的不同反应作为支原体存在的函数，对于上呼吸消化道体外实验的翻译是潜在的问题。目前尚不清楚这些发现是否代表污染或这种特殊组织的基线状态。
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. 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. 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. 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. 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. 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.
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影响因子：
2.6
作者：
Chhetri, Dinesh K.;Berke, Gerald S.
通讯作者：
Berke, Gerald S.
影响因子：
4.6
作者：
Nakamura R;Mukudai S;Bing R;Garabedian MJ;Branski RC
通讯作者：
Branski RC
影响因子：
7.3
作者：
Akasaka, Yoshikiyo;Ono, Ichiro;Ishii, Toshiharu
通讯作者：
Ishii, Toshiharu
影响因子：
1.4
作者：
Branski, Ryan C.;Barbieri, Silvia S.;Felsen, Diane
通讯作者：
Felsen, Diane
DOI：
10.1111/j.1574-695x.1998.tb01212.x
发表时间：
1998-11-01
影响因子：
--
作者：
Watanabe, T;Shibata, K;Totsuka, Y
通讯作者：
Totsuka, Y