Massachusetts Eye and Ear/ Harvard Initiative for the NIDCD National Human Ear Resource Network

马萨诸塞州眼与耳/哈佛 NIDCD 国家人耳资源网络倡议

• 批准号: 10705823
• 负责人: M. Charles Liberman
• 金额: $ 59.5万
• 依托单位: MASSACHUSETTS EYE AND EAR INFIRMARY
• 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 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10705823
• 关键词: Acceleration; Adopted; Archives; Auditory; Autopsy; Biological; Biopsy; Cell Survival; Celloidin; Chemicals; Cochlea; Collaborations; Collection; Complex; Coupling; DNA; DNA Fragmentation; Data; Databases; Decalcification; Development; Disease; Documentation; Ear; Education and Outreach; Educational Curriculum; Educational process of instructing; Educational workshop; Equilibrium; Etiology; Excision; External Ear; Eye; Fellowship; Fellowship Program; Genetic; Genetic study; Genomics; Hearing problem; Heavy Metals; Hematoxylin and Eosin Staining Method; Hereditary Disease; Histopathology; Human; Hybrids; Image; Impairment; Improve Access; In Situ Hybridization; Inherited; Institution; Instruction; International; Laboratories; Labyrinth; Lasers; Libraries; Life; Machine Learning; Manuals; Massachusetts; Medical; Medical Records; Mentors; Methods; Methylmethacrylate; Microtomy; Modification; Molecular; Molecular Analysis; Morphology; Muscle; National Institute on Deafness and Other Communication Disorders; Online Systems; Osmium; Pathogenicity; Pathologic; Pathology; Pattern; Plastic Embedding; Preparation; Process; Proteins; Protocols documentation; RNA; Recording of previous events; Registries; Research; Research Personnel; Residencies; Resolution; Resources; Sampling; Scanning; Science; Scientist; Slide; Specimen; Speed; Stains; Structure; Surveys; Techniques; Temporal bone structure; Three-Dimensional Image; Tissue Donors; Tissue Sample; Tissues; Training; Underrepresented Minority; United States; Variant; Work; X-Ray Tomography; algorithm development; bone; cell injury; data de-identification; deafness; digital; equilibration disorder; ex vivo imaging; exome sequencing; experience; gene therapy; genetic analysis; genetic information; hereditary hearing loss; improved; insight; knowledge base; light microscopy; meetings; middle ear; new technology; next generation; non-invasive imaging; novel; novel strategies; online tutorial; otoconia; outreach; preservation; programs; public health relevance; reconstruction; sample fixation; searchable database; skills; software development; success; symposium; therapy development; tool; translational study; virtual; web portal

Project Summary This proposal to join the NIDCD National Human Ear Resource Network pools the skills of an international consortium of researchers with decades of experience in the histopathological study of acquired and hereditary disorders of hearing and balance, and in the curation of one of the largest collections of human temporal-bone (TB) specimens in the world. Because the ear cannot be biopsied, and non-invasive imaging cannot yet provide cellular level resolution, insight into the pathologies underlying deafness and balance disorders derives from the study of TBs removed at autopsy, which classically have been decalcified, plastic-embedded and serially sectioned, with a subset then stained with hematoxylin and eosin (H&E) for pathological study. In Aim 1, we continue to accrue and process human TBs, document the histopathology and medical/otologic history, and upload the de-identified data to a searchable database, along with digitized images of all the H&E-stained slide sets, to jumpstart more widespread collaborations in the study of human otopathology. We will also continue to distribute unstained sections from our TB archives to researchers for additional morphological, molecular biological or genetic studies of particular disorders. In Aim 2, we enhance techniques for human TB study in three key ways: first (Aim 2a), by perfecting a novel approach to embedding (methyl methacrylate) and sectioning (laser microtomy) that is cheaper and faster than celloidin, allows morphological and chemical analysis of undecalcified bone and otoconia, and is less destructive to protein and RNA/DNA; second (Aim 2b), by pursuing the ex vivo imaging of intact cochleas via high-resolution x-ray tomography, enhanced by coupling celltype-specific immunomarkers to heavy metals and by developing software to automate extraction of quantitative histopathological data from these 3D image stacks; and third (Aim 2c), by developing and applying techniques for DNA extraction and whole-exome sequencing from donor tissue samples and/or celloidin TBs, to identify the pathogenic variants responsible for the numerous hereditary hearing loss cases in our archives that were not genotyped during life. In Aim 3, we strive to accelerate the pace of discovery in human otopathology by sponsoring fellowships, organizing national symposia and by creating and disseminating training materials, via hands-on courses, online tutorials, technical manuals on the classic and novel techniques needed for the procurement, processing and histopathological analysis of human temporal bones.

项目摘要 这项加入NIDCD国家人耳资源网络的提议汇集了一名国际 在获得性和遗传性组织病理学研究方面有数十年经验的研究人员组成的联盟 听力和平衡障碍，以及人类最大的颞骨收藏之一的管理 (TB)世界标本。因为耳朵不能做活组织检查，非侵入性成像也不能 提供细胞水平的分辨率，洞察耳聋和平衡障碍的潜在病理 从尸检时取出的TBS的研究中，经典的TBS已被脱钙，塑料嵌入和 连续切片，取一组标本，苏木精-伊红(H&E)染色，作病理观察。 在目标1中，我们继续积累和处理人类TBS，记录组织病理学和 医疗/耳科病史，并将未识别的数据与数字化的数据一起上传到可搜索的数据库 所有H&E染色幻灯片的图像，以启动在人类研究中更广泛的合作 耳科病理学。我们还将继续从我们的结核病档案中分发未染色的切片给研究人员 特殊疾病的其他形态、分子生物学或遗传学研究。 在目标2中，我们在三个主要方面加强了人类结核病研究的技术：第一(目标2a)，通过完善 包埋(甲基丙烯酸甲酯)和切片(激光显微切割)的新方法，成本更低， 比火棉胶快，允许对未脱钙的骨骼和耳石进行形态和化学分析，并且是 对蛋白质和RNA/DNA的破坏性较小；第二(目标2b)，通过对完整的 通过高分辨率x射线断层扫描，通过将细胞类型特异性免疫标记物偶联到 并通过开发软件自动提取定量组织病理学数据 这些3D图像堆栈；第三(目标2c)，开发和应用DNA提取和 从供体组织样本和/或火棉蛋白TBS中进行全外显子组测序，以鉴定致病变异 对我们档案中的许多遗传性耳聋病例负有责任，这些病例在有生之年没有进行基因分型。 在目标3中，我们努力通过赞助奖学金来加快人类耳科病理学的发现步伐， 组织国家专题讨论会，并通过实践课程编写和分发培训材料， 关于采购、加工所需的经典技术和新技术的在线教程、技术手册 以及人类颞骨的组织病理学分析。