Detection of BDNF-Related Proteins in Human Perilymph in Patients With Hearing Loss
Detection of BDNF-Related Proteins in Human Perilymph in Patients With Hearing Loss
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DOI：
10.3389/fnins.2019.00214
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发表时间：
2019-03-26
影响因子：
4.3
通讯作者：
Wamecke, Athanasia
中科院分区：
文献类型：
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作者：
de Vries, Ines;Schmitt, Heike;Wamecke, Athanasia
人工耳蜗植入的结果取决于多个变量，包括耳蜗的基础健康状况。脑源性神经营养因子（BDNF）已被证明支持螺旋神经节神经元，并在动物模型中改善植入功能。内源性BDNF或BDNF调节蛋白是否可以作为生物标志物来预测耳蜗健康和植入结果尚未研究。在颅底手术期间从正常听力患者（n = 3）获得的人耳蜗组织中鉴定BDNF和下游信号分子的基因表达。基于基因表达数据，利用生物信息学分析预测BDNF对蛋白质的调控。通过纳米级液相色谱串联质谱法（纳米LC-MS/ MS），研究了在耳蜗植入或颅底手术切除前庭神经鞘瘤期间从听力受损患者（n = 38）获得的外淋巴液（n = 41）中对应于这些基因的蛋白质的存在。通过质谱法分析了41个外淋巴样品，尽管有三个患者接受双侧人工耳蜗植入。这些特定的BDNF调节蛋白在任何外淋巴样品中都检测不到。随后，外淋巴蛋白质组数据与损伤途径分析（IPA）的有针对性的分析，确定了进一步的蛋白质在人类外淋巴液中，可以由BDNF调节。这些BDNF调节蛋白与植入前残余听力（RH）的存在以及1年后人工耳蜗植入的性能数据相关。总体而言，与部分RH患者相比，重度听力受损患者的BDNF调节蛋白表达水平降低。磷脂转运蛋白与患者术前听力水平呈正相关。我们的数据表明，基因表达阵列和生物信息学分析的组合可以帮助预测与BDNF通路相关的下游信号蛋白。外淋巴液的蛋白质组学分析可能有助于确定这些分子在患病器官中的存在或不存在。这种预测算法对诊断和治疗的影响需要在进一步的研究中建立。
The outcome of cochlear implantation depends on multiple variables including the underlying health of the cochlea. Brain derived neurotrophic factor (BDNF) has been shown to support spiral ganglion neurons and to improve implant function in animal models. Whether endogenous BDNF or BDNF-regulated proteins can be used as biomarkers to predict cochlear health and implant outcome has not been investigated yet. Gene expression of BDNF and downstream signaling molecules were identified in tissue of human cochleae obtained from normal hearing patients (n = 3) during skull base surgeries. Based on the gene expression data, bioinformatic analysis was utilized to predict the regulation of proteins by BDNF. The presence of proteins corresponding to these genes was investigated in perilymph (n = 41) obtained from hearing-impaired patients (n = 38) during cochlear implantation or skull base surgery for removal of vestibular schwannoma by nanoscale liquid chromatography coupled to tandem mass spectrometry (nano LC-MS/ MS). Analyzed by mass spectrometry were 41 perilymph samples despite three patients undergoing bilateral cochlear implantation. These particular BDNF regulated proteins were not detectable in any of the perilymph samples. Subsequently, targeted analysis of the perilymph proteome data with Ingenuity Pathway Analysis (IPA) identified further proteins in human perilymph that could be regulated by BDNF. These BDNF regulated proteins were correlated to the presence of residual hearing (RH) prior to implantation and to the performance data with the cochlear implant after 1 year. There was overall a decreased level of expression of BDNF-regulated proteins in profoundly hearing-impaired patients compared to patients with some RH. Phospholipid transfer protein was positively correlated to the preoperative hearing level of the patients. Our data show that combination of gene expression arrays and bioinformatic analysis can aid in the prediction of downstream signaling proteins related to the BDNF pathway. Proteomic analysis of perilymph may help to identify the presence or absence of these molecules in the diseased organ. The impact of such prediction algorithms on diagnosis and treatment needs to be established in further studies.