Molecular etiology of virus-induced sensorineural hearing loss

病毒引起的感音神经性听力损失的分子病因学

• 批准号: 10705840
• 负责人: Marie Kubota
• 金额: $ 19.74万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-16 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10705840
• 关键词: Acute; Address; Affect; Anatomy; Animal Model; Antibodies; Antiviral Agents; Apical; Apoptosis; Apoptotic; Auditory; Auditory Brainstem Responses; Auditory system; Betaherpesvirinae; Biological Assay; Biology; Birth; C57BL/6 Mouse; Cell Count; Cells; Chronic; Cochlea; Collaborations; Confocal Microscopy; Cytomegalovirus; Cytomegalovirus Infections; Development; Disease; Double Stranded DNA Virus; Embryonic Development; Equipment and supply inventories; Etiology; Evaluation; Experimental Animal Model; Family; Foundations; Future; Gangliosides; Generations; Goals; Hair Cells; Herpesviridae; Histocytochemistry; Histopathology; Human; IFNAR1 gene; Immune; Immune response; Inbred BALB C Mice; Infection; Injections; Inner Ear Infection; Integration Host Factors; Interferon Type I; Knowledge; Labyrinth; Lassa virus; Lead; Lesion; Location; Measures; Medical; Methods; Modeling; Molecular; Mumps virus; Murid herpesvirus 1; Mus; Neonatal; Neuroglia; Neurons; Paramyxovirus; Pathogenesis; Pathologic; Pathology; Patients; Predisposition; Prevention; Preventive treatment; Principal Investigator; Protocols documentation; RNA Viruses; Reporting; Research; Research Personnel; Research Training; Rodent; Role; Scientist; Sensorineural Hearing Loss; Steroids; Structure of posterior semicircular canal; Supporting Cell; Systemic infection; Testing; Texas; Tissues; Universities; Viral; Viral Pathogenesis; Viral Proteins; Viral load measurement; Virulence; Virus; Virus Diseases; Virus Receptors; Work; animal model development; base; biomarker identification; career; cell injury; cell type; experience; ganglion cell; hearing impairment; immune cell infiltrate; improved outcome; in vivo; in vivo Model; interferon alpha receptor; mouse model; neonatal mice; novel; otoacoustic emission; ototoxicity; pathogen; postnatal; spiral ganglion; viral detection; virology

PROJECT ABSTRACT Viral infection is a common cause of sensorineural hearing loss (SNHL). However, the underlying mechanism is poorly understood. For most viruses, it is unclear whether SNHL is caused by direct viral infection of inner ear tissue or by a secondary effect through an immune response, or a combination of both. Antiviral drugs and steroids only improve the outcome for a small group of patients. It is therefore imperative to understand the molecular etiology of virus-induced SNHL for the development of effective preventions and treatments. The research proposes a systemic evaluation of viral effects on the inner ear using mumps virus (MuV), a human virus that causes SNHL after birth (acquired SNHL), and murine cytomegalovirus (mCMV) to understand human CMV disease that causes SNHL during embryonic development (congenital SNHL). There is no small animal model to assess SNHL caused by MuV. In Aim1, it is proposed to establish novel in vivo mouse models to assess how mCMV and MuV infections affect inner ear cells. Infection will be done locally and controlled via the posterior semicircular canal. The onset and extent of hearing loss will be measured by auditory brainstem response and distortion product otoacoustic emissions. Histochemistry will quantify apoptotic cells, virus-infected cells, and immune cells in defined anatomical locations of the cochlea from apex to base. Because the model is expected to be highly controllable, it will be optimized so that the detrimental mechanisms of different viruses can be compared to advance the mechanistic understanding of virus-induced SNHL. Aim2 focuses on understanding the etiology of the tissue damage -direct viral infection or immune response- in mCMV- and MuV- administrated cochleae, and to analyze the mechanisms at the cellular and molecular level. Susceptible cell subtypes will be quantified in P2 and P21 mice injected with mCMV and MuV before and after the onset of hearing loss by using known and recently identified markers. These include new pericochlear cell subtypes in the neonatal cochlea that were recently reported by the principal investigator (PI), as well as the new subtypes of type I spiral ganglion cells. The expression of a ganglioside called GM2, a MuV receptor which the PI identified in a previous study, and various viral host factors will be also quantified in each cochlear cell subtype to understand their correlation with cellular damage. Further, the types and numbers of infiltrating immune cells post PSC injection of mCMV and MuV in P2 and P21 mice will be assessed with flow cytometric analysis with cell type-specific markers. Together, completion of these aims will provide essential details of the mechanism of SNHL caused by mCMV and MuV, and will establish a protocol that can be applied to other hearing loss-causing viruses such as Lassa virus, which is not suitable for animal model development because of its high biosafety level. The PI envisions this project as the first step of her future research journey as a clinician-scientist with a focus on prevention and treatment of viral-induced SNHL.

项目摘要 病毒感染是感音神经性听力损失（SNHL）的常见原因。然而，其根本机制是 不太了解。对于大多数病毒来说，尚不清楚SNHL是否是由内耳病毒直接感染引起的 组织或通过免疫反应产生的次级效应，或两者的组合。抗病毒药物和 类固醇只能改善一小部分患者的治疗结果。因此，有必要了解 病毒引起的 SNHL 的分子病因学，以开发有效的预防和治疗方法。这 研究提出使用人类腮腺炎病毒（MuV）对病毒对内耳的影响进行系统评估 导致出生后 SNHL（获得性 SNHL）的病毒和了解人类的鼠巨细胞病毒 (mCMV) 在胚胎发育过程中导致 SNHL（先天性 SNHL）的 CMV 疾病。没有小动物 评估 MuV 引起的 SNHL 的模型。在 Aim1 中，建议建立新型体内小鼠模型 评估 mCMV 和 MuV 感染如何影响内耳细胞。感染将在本地进行并通过 后半规管。听力损失的发生和程度将通过听觉脑干来测量 响应和失真产物耳声发射。组织化学将量化病毒感染的凋亡细胞 细胞和免疫细胞位于耳蜗从顶端到基部的特定解剖位置。 因为模型是 预计高度可控，会进行优化，使得不同病毒的有害机制 可以进行比较，以推进对病毒诱导的 SNHL 机制的理解。目标2重点关注 了解 mCMV- 和 MuV- 组织损伤的病因 - 直接病毒感染或免疫反应 - 管理耳蜗，并在细胞和分子水平上分析其机制。易感细胞 将在发病前后注射 mCMV 和 MuV 的 P2 和 P21 小鼠中对亚型进行量化 使用已知和最近识别的标记导致听力损失。其中包括新的耳蜗周围细胞亚型 主要研究者（PI）最近报告的新生儿耳蜗，以及新的亚型 I 型螺旋神经节细胞。一种称为 GM2 的神经节苷脂的表达，这是 PI 鉴定出的一种 MuV 受体 在之前的一项研究中，各种病毒宿主因子也将在每个耳蜗细胞亚型中进行量化，以 了解它们与细胞损伤的相关性。此外，浸润免疫细胞的类型和数量 PSC 在 P2 和 P21 小鼠中注射 mCMV 和 MuV 后，将通过流式细胞术分析进行评估 细胞类型特异性标记。总之，完成这些目标将提供机制的基本细节 由 mCMV 和 MuV 引起的 SNHL，并将建立可应用于其他导致听力损失的协议 拉沙病毒等病毒生物安全性高，不适合动物模型开发 等级。 PI 将该项目视为她作为一名临床医生科学家未来研究之旅的第一步 重点关注病毒性SNHL的预防和治疗。