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.

项目摘要加入NIDCD国家人耳资源网络的这一建议汇集了国际的技能在获得和遗传的组织病理学研究中拥有数十年经验的研究人员的财团听力和平衡的疾病，以及最大的人类颞骨收藏之一的策划（TB）世界上的标本。因为耳朵不能进行活检，并且非侵入性成像还不能提供细胞水平的分辨率，对聋哑的病理的洞察力和平衡疾病衍生根据对尸检中除去的TBS的研究，该研究经典被脱钙，塑料包裹和串行分段，然后用苏木精和曙红（H＆E）染色的子集进行病理研究。在AIM 1中，我们继续累积和处理人类TB，记录组织病理学和医学/耳科历史记录，并将去识别的数据上传到可搜索的数据库，以及数字化所有H＆E染色幻灯片集的图像，以开始在人类研究中更广泛的合作眼科病理学。我们还将继续向研究人员分发未染色的部分，以供研究人员特定疾病的其他形态学，分子生物学或遗传研究。在AIM 2中，我们通过三个关键方式来增强人类结核病研究的技术：首先（AIM 2A），完善新颖的嵌入方法（甲基丙烯酸甲酯）和切片（激光显微切开术），便宜且比细胞素快，可以对未钙化的骨骼和耳尾症进行形态学和化学分析，并且IS 对蛋白质和RNA/DNA的破坏性较小；第二（AIM 2B），通过追求完整的离体成像通过高分辨率X射线断层扫描的耳蜗，通过将细胞类型特异性免疫标记耦合到增强重金属和开发软件以自动从中提取定量组织病理学数据这些3D图像堆栈；和第三（AIM 2C），通过开发和应用用于DNA提取的技术和从供体组织样品和/或细胞素TBS进行的全外观测序，以鉴定致病性变异负责我们档案中众多的遗传性听力损失案例，这些档案中未在生命中进行基因分型。在AIM 3中，我们努力通过赞助奖学金，加速人类眼病理学的发现速度，通过动手课程组织和传播培训材料，组织和传播培训材料，在线教程，有关采购所需的经典和新颖技术的技术手册和人类颞骨的组织病理学分析。