Ear Level Cooling for Otoprotection

摘要 我们以前证明了显着的保护顺铂诱导的听力损失，使用冷水耳 渠灌然而，该研究仅限于单次推注顺铂和急性时间段 （Spankovich等人，2016）。最近，我们研究了耳道局部冷却的应用， 顺铂的重复剂量，在一个扩大的时间段，并使用替代的冷却方法（耳 由Peltier装置冷却的棒）。在这里，我们建议推进我们的局部耳冷却的新应用， 缓解药物引起的听力损失;我们称之为Cool OtOprotective ear Lumen（COOL）疗法。 假设：我们假设（1）我们的COOL疗法将在大鼠模型中显示保护作用;（2）保护作用 从我们的COOL疗法是介导的减少耳蜗摄取顺铂，表达或冷休克 蛋白质，并减少上游和下游细胞凋亡细胞死亡途径;（3）我们将观察到 保护前庭损伤与我们的COOL疗法。 具体目标：目标1。为了确定COOL治疗是否减少顺铂摄取和前庭神经病变， 病理学，增加编码冷休克蛋白的转录本的表达，并减少细胞凋亡。 压力路径。 研究设计：对于目标1，豚鼠和Long Evans大鼠在接受顺铂治疗时将暴露于顺铂。 COOL治疗，并与假手术对照组和温热动物进行比较。目标1（a）将决定我们的研究结果是否 翻译到另一个物种（大鼠），并探索前庭效应。目的1（B）检测抗体 顺铂DNA加合物的免疫染色，质谱法测量耳蜗中的铂浓度 组织和用荧光标记的顺铂处理的动物的共聚焦图像。目标1（c，d）将审查 冷休克蛋白的表达以及脂质过氧化和半胱天冬酶-3激活的证据。

ABSTRACT We previously demonstrated significant protection from cisplatin induced hearing loss using cool water ear canal irrigation. However, the study was limited to a single bolus injection of cisplatin and acute time period (Spankovich et al. 2016). More recently, we examined application of localized cooling of the ear canal with repeated doses of cisplatin, over an expanded period of time, and using an alternative method of cooling (ear bar cooled by a Peltier device). Here, we propose to advance our novel application of localized ear cooling in mitigation of drug induced hearing loss; which we call Cool OtOprotective ear Lumen (COOL) therapy. Hypotheses: We hypothesize that (1) our COOL therapy will show protection in a rat model; (2) protection from our COOL therapy is mediated by reduced cochlear uptake of cisplatin, expression or cold shock proteins, and reduced upstream and downstream cell apoptotic cell death pathways; (3) that we will observe protection from vestibular damage with our COOL therapy. Specific Aims: Aim 1. To determine whether COOL therapy reduces cisplatin uptake and auditory-vestibular pathology, increases the expression of transcripts that code for cold-shock proteins, and reduced cellular stress pathways. Study Design: For Aim 1, Guinea pig and Long Evans rats will be exposed to cisplatin while undergoing COOL therapy and compared to sham controls and warmed animals. Aim 1(a) will determine if our findings translate to another species (rat) and explore vestibular effects. Aim 1(b) will examine antibody immunostaining for cisplatin DNA-adducts, mass spectrometry measured platinum concentrations in cochlear tissue, and confocal images of animals treated with fluorescent tagged cisplatin. Aim 1(c,d) will examine expression of cold shock proteins and evidence of lipid perioxidation and caspase-3 activation.