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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要加入 NIDCD 国家人耳资源网络的提议汇集了国际技术人员的技能由在获得性和遗传性组织病理学研究方面拥有数十年经验的研究人员组成的联盟听力和平衡障碍，以及最大的人类颞骨收藏之一的策展（TB）世界各地的标本。因为耳朵无法进行活检，而且无创成像还无法提供细胞水平的分辨率，深入了解耳聋和平衡障碍的病理学来自尸检时取出的结核病的研究，这些结核病通常经过脱钙、塑料嵌入和连续切片，然后用苏木精和伊红 (H&E) 染色进行病理研究。在目标 1 中，我们继续收集和处理人类结核病，记录组织病理学和医学/耳科病史，并将去识别化的数据连同数字化数据一起上传到可搜索的数据库所有 H&E 染色幻灯片组的图像，以推动人类研究中更广泛的合作耳病理学。我们还将继续将结核病档案中未染色的部分分发给研究人员，以供研究人员使用。对特定疾病的额外形态学、分子生物学或遗传学研究。在目标 2 中，我们通过三个关键方式增强人类结核病研究技术：首先（目标 2a），通过完善包埋（甲基丙烯酸甲酯）和切片（激光切片）的新颖方法，成本更低，比赛璐珞更快，可以对未脱钙骨和耳石进行形态和化学分析，并且对蛋白质和 RNA/DNA 的破坏性较小；第二（目标 2b），通过追求完整的离体成像通过高分辨率 X 射线断层扫描对耳蜗进行观察，并通过将细胞类型特异性免疫标记物与重金属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），通过开发和应用 DNA 提取技术对供体组织样本和/或赛璐珞结核进行全外显子组测序，以鉴定致病变异我们档案中的许多遗传性听力损失病例在生前没有进行基因分型。在目标 3 中，我们努力通过赞助奖学金来加快人类耳病理学的发现步伐，组织全国研讨会并通过实践课程制作和传播培训材料，采购、加工所需的经典和新颖技术的在线教程、技术手册以及人类颞骨的组织病理学分析。