High resolution 3D mapping of cellular heterogeneity within multiple types of mineralized tissues

多种矿化组织内细胞异质性的高分辨率 3D 绘图

• 批准号: 10700252
• 负责人: Martin K Lotz
• 金额: $ 53.5万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-21 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10700252
• 关键词: 3-Dimensional; Affect; Biology; Cartilage; Cells; Characteristics; Collaborations; Communication; Communities; Confocal Microscopy; Data; Data Analyses; Data Files; Databases; Discipline; Educational workshop; Ensure; Environment; Faculty; Gene Expression Profile; Genetic; Genetic Transcription; Goals; Grant; Guidelines; Health; Heterogeneity; Histologic; Histology; Human; Human BioMolecular Atlas Program; Human Biology; Human Resources; Image; In Situ; In Situ Hybridization; Individual; Joints; Knee; Ligands; Maps; Mechanics; Methods; Minerals; Molecular; Molecular Biology; Normalcy; Outcome; Pathway interactions; Pattern; Process; Protocols documentation; Quality Control; Rare Diseases; Research; Research Personnel; Resolution; Resources; Role; Sampling; Signal Pathway; Signal Transduction; Skeletal system; Skeleton; Source; Structure; Systems Biology; Techniques; Technology; Therapeutic; Tissue imaging; Tissues; Tooth Tissue; Tooth structure; Training; Transcript; Translating; Videoconferencing; Visual; base; bone; cell type; experience; histological image; human tissue; insight; interest; mechanical load; meetings; member; multimodality; outreach; programs; receptor; response; skeletal; skeletal tissue; soft tissue; substantia spongiosa; three dimensional structure; tissue mapping; transcription factor; transcriptome; transcriptomics; virtual; visual map

Overall Abstract: The goal of the HuBMAP program is to appreciate the unique contextual role of individual cells within the 3D structure of a tissue at its most basic level of transcriptional activity, cellular signaling and cellular response. To date the subject tissues have not included the mineralized skeletal system due to technical issues that preclude the requirements of the HuBMAP program. We have solved those issues with a protocol that is capable of performing multimodal histology that include methods for advanced and repetitive in situ hybridization. We will develop this technology for three very different mineralized skeletal tissues: tooth, trabecular bone and cartilage structures of the knee. The Coordination Core will acquire the three tissue types from de-identified human sources. Each sample will be oriented to its source tissue, imaged by µCT to capture its mineral structure and processed into a histological stack to create a 3D representation of the tissue. Using the histological stack, the Mineralized Tissue Program will perform both in situ hybridization to identify the multiple cell types and seqFISH hybridization to capture the cell transcriptome of the identified cell types. These technically demanding steps will require direction from established HuBMAP investigators as well as UConn faculty who are experts in high resolution confocal microscopy. The Data Analysis Core will translate the image files generated by these techniques into 3D cellular maps of the target tissue and transcriptome composition of each cell type. From those data files, our contextual molecular mapping program, TOPAS, will examine the transcriptional and signaling pathways to impute how neighboring cells coordinate their activities to respond to mechanical loading and systemic factors that regulate skeletal health. The workflow and analytical platforms that we developed for the skeletal system will be aligned with the requirement of the HIVE including an outreach initiative. First, we will provide opportunities to transfer the histological technology to major academic skeletal research groups. Second, once our skeletal data becomes available from the central HIVE source, we will develop virtual workshops to inform the skeletal biology community of this valuable resource and how it can be utilized to unravel rare diseases affecting the skeleton. HuBMAP will be a transformational technology that every tissue centric group needs to incorporate. Our role is to ensure that the skeletal biology community is included in this new experimental platform, and that it is employed to solve the major genetic and therapeutic challenges affecting skeletal health.

总体摘要： Hubmap程序的目标是欣赏3D中各个单元的独特上下文作用 组织在其最基本的转录活性，细胞信号传导和细胞反应下的结构。 迄今为止，由于技术问题，主题组织尚未包括矿物化的骨骼系统 排除Hubmap程序的要求。我们已经用协议解决了这些问题 能够执行包括高级和重复原位方法的多模式组织学 杂交。我们将针对三个非常不同的矿化骨骼时机开发这项技术：牙齿， 膝盖的小梁骨和软骨结构。协调核心将获取三种组织类型 来自被识别的人类来源。每个样品将定向其源组织，由µCT成像以捕获 它的矿物结构并加工成组织学堆栈以创建组织的3D表示。使用 组织学堆栈，矿化组织程序将同时执行原位杂交，以确定 多种细胞类型和Seq Fish杂交以捕获已鉴定的细胞类型的细胞转录组。 这些技术要求的步骤将需要既定的Hubmap调查人员以及 UConn教师是高分辨率共聚焦显微镜专家。数据分析核心将翻译 这些技术生成的图像文件成目标组织的3D蜂窝图和转录组 每种细胞类型的组成。从这些数据文件中，我们的上下文分子映射程序TOPA将 检查转录和信号传导途径，以指示相邻细胞如何协调其活性 应对调节骨骼健康的机械负荷和系统因素。工作流和 我们为骨骼系统开发的分析平台将与Hive的需求保持一致 包括一项宣传计划。首先，我们将提供将组织学技术转移到的机会 主要的学术骨骼研究小组。其次，一旦我们的骨骼数据从中央获得 蜂巢来源，我们将开展虚拟研讨会，以告知骨骼生物学社区此价值 资源以及如何利用它来揭示影响骨骼的罕见疾病。 hubmap将是 每个以组织为中心的组织都需要合并的变革性技术。我们的角色是确保 骨骼生物学界包括在这个新的实验平台中，并携带 影响骨骼健康的主要遗传和治疗挑战。