Zika Virus Cell Tropism and Pathogenesis in the Developing Inner Ear

黄病毒家族的寨卡病毒(ZIKV)对新生儿的健康构成重大威胁 婴幼儿。寨卡病毒感染主要影响神经系统的发育和其 神经官能症已经根深蒂固。2016年9月，ZIKV被认为与 出生前接触过的新生儿，但基本上对发育情况一无所知 病理存在于听觉通路中。寨卡病毒暴露婴儿耳声发射异常的报道 提示至少部分感觉神经缺陷可能起源于耳蜗区。 内耳。这项研究将集中在胚胎和围产期ZIKV暴露时间的相关性。 使用两个动物模型(鸡和小鼠)，对感染的内耳细胞的类型进行研究。初步 资料显示，ZIKV可感染耳蜗区，特别是感觉区、血管纹(被盖) 鸡耳蜗内的血管)和听神经节。拟议的研究将涉及交付ZIKV 直接到胚胎内耳和耳蜗处评估细胞趋向性和耳聋相关的发病机制 和ZIKV一起。目的1通过直接注射确定ZIKV在卵子中的细胞趋向性和感染时间 病毒从胚胎3-5天进入鸡耳囊。病毒的分布、细胞增殖、细胞凋亡和 与先天免疫反应相关的基因表达将在感染后几天进行检测。目标2将 ZIKV在胚胎(E12.5、E15.5)和新生儿体内体外感染的细胞嗜性和时间的确定 (出生后第1天)小鼠耳蜗培养。毛细胞、支持细胞或中间细胞的遗传标记 在长达6天的培养中，血管纹将有助于跟随这些细胞类型的命运。病毒传播， 细胞增殖、细胞凋亡、毛细胞超微结构和毛细胞机械转导将在几个方面进行评估 在接触ZIKV几天后。将通过抗体阻断实验来探索骨膜期的作用 间充质作为ZIKV的直接目标。这些研究应该有助于理解寨卡病毒感染是如何 感觉神经末梢的发育可能导致婴儿的感音神经性听力损失。

The Zika virus (ZIKV) of the flaviviridae family of viruses poses a significant threat to the health of newborn infants and young children. ZIKV infection predominantly affects the development of the nervous system and its neurotopism has been well-established. In September 2016, ZIKV was linked to sensorineural hearing loss in newborn infants that were prenatally exposed, but essentially nothing is known about where the developmental pathologies lie in the auditory pathway. Reports of abnormal otoacoustic emissions in ZIKV-exposed infants suggest that at least some of the sensorineural defects may originate within the cochlea, the auditory organ of the inner ear. This study will focus on correlating the timing of ZIKV exposure at embryonic and perinatal stages to the types of inner ear cells that become infected, using two animal models (chicken and mice). Preliminary data show that ZIKV can infect the cochlea, in particular the sensory domain, the stria vascularis (tegmentum vasculosum in the chicken cochlea) and the auditory ganglion. The proposed studies will involve delivery of ZIKV directly to the embryonic inner ear and the cochlea to assess cellular tropism and otic pathogenesis associated with ZIKV. Aim 1 will determine the cellular tropism and timing of ZIKV infection in ovo by direction injection of virus into the chicken otocyst from embryonic day (E) 3–5. Virus distribution, cell proliferation, apoptosis, and gene expression related to innate immune responses will be assayed several days after infection. Aim 2 will determine the cellular tropism and timing of ZIKV infection in vitro in embryonic (E12.5, E15.5) and neonatal (postnatal day 1) mouse cochlea cultures. Genetic labeling of hair cells, supporting cells, or intermediate cells of the stria vascularis will aid in following the fates of these cell types for up to 6 days in culture. Virus distribution, cell proliferation, apoptosis, hair cell ultrastructure, and hair cell mechanotransduction will be evaluated several days after ZIKV exposure. Antibody blocking experiments will be used to explore the role of the periotic mesenchyme as a direct target of ZIKV. These studies should aid in understanding how ZIKV infection of the developing sensory periphery might contribute to sensorineural hearing loss in human infants.