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 计划的目标是了解 3D 细胞中单个细胞的独特背景作用 组织在转录活性、细胞信号传导和细胞反应等最基本水平上的结构。 迄今为止，由于技术问题，主题组织尚未包括矿化骨骼系统 我们已经通过协议解决了这些问题。 能够执行多模式组织学，包括高级和重复原位方法 我们将为三种截然不同的矿化骨骼组织开发这项技术：牙齿、 膝盖的小梁骨和软骨结构将获得三种组织类型。 每个样本都将定向到其源组织，并通过 µCT 成像来捕获。 其矿物结构并加工成组织学堆栈，以创建组织的 3D 表示。 组织学堆栈中，矿化组织计划将执行原位杂交来识别 多种细胞类型和 seqFISH 杂交以捕获已识别细胞类型的细胞转录组。 这些技术要求较高的步骤将需要现有 HuBMAP 研究人员以及 康涅狄格大学高分辨率共聚焦显微镜专家的教师将负责翻译。 将这些技术生成的图像文件转化为目标组织和转录组的 3D 细胞图 我们的上下文分子作图程序 TOPAS 将根据这些数据文件来分析每种细胞类型的组成。 检查转录和信号传导途径，以估算邻近细胞如何协调其活动 响应机械负荷和调节骨骼健康的系统因素。 我们为骨骼系统开发的分析平台将符合 HIVE 的要求 首先，我们将提供将组织学技术转让给其他国家的机会。 其次，一旦我们的骨骼数据从中央获得。 HIVE 来源，我们将开发虚拟研讨会，向骨骼生物学界通报这一宝贵的信息 资源以及如何利用它来解决影响骨骼的罕见疾病将是一个。 每个以组织为中心的团体都需要采用变革性技术，我们的作用是确保 骨骼生物学界包含在这个新的实验平台中，它被用来解决 影响骨骼健康的主要遗传和治疗挑战。