Organ Specific Project - Bone Marrow

器官特定项目 - 骨髓

• 批准号: 10118853
• 负责人: KATHRIN M BERNT
• 金额: $ 44.04万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10118853
• 关键词: 3-Dimensional; Address; Adipocytes; Aging; Anatomy; Archives; Autopsy; Biological Assay; Biology; Blood Cells; Blood Vessels; Bone Banks; Bone Growth; Bone Marrow; Bone Marrow Cells; Bone Tissue; Cell Nucleus; Cells; Cellular Assay; Chromatin; Clinical; Collection; Communities; Complex; Consent; Data; Detection; Development; Disease; Doctor of Philosophy; Endothelium; Epidemiology; Epigenetic Process; Extracellular Matrix; Female; Fluorescent in Situ Hybridization; Future; Genetic Transcription; Genomics; Goals; Heart; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; Histologic; Homeostasis; Human; Human BioMolecular Atlas Program; Image; Infrastructure; Knowledge; Leadership; Location; Longevity; Lymphoid; Mesenchymal; Mesenchymal Stem Cells; Metadata; Minerals; Multiomic Data; Myelogenous; Organ; Osteoblasts; Osteocytes; Osteogenesis; Persons; Pilot Projects; Play; Proteomics; Protocols documentation; Regulator Genes; Research; Resolution; Role; Sampling; Series; Signal Pathway; Small Nuclear RNA; Specimen; Sternum; Stromal Cells; System; Three-Dimensional Imaging; Tissues; Transposase; age group; base; biobank; bone; cell type; cellular imaging; design; epigenome; experience; indexing; inter-individual variation; male; mechanical properties; member; microCT; multiple omics; progenitor; programs; serial imaging; transcriptome; transcriptome sequencing; transcriptomics

PROJECT SUMMARY – BONE ORGAN SPECIFIC PROJECT The bone is a highly complex organ made of three tissue types with distinct functions, including cells from hematopoietic, mesenchymal and endothelial lineages. Confined within one organ and thoroughly mixed with each other, these three tissues are mutually dependent for their development and homeostasis. The spatial organization of the various cell types and their cross-talk in the bone is a major gap of our knowledge. The goal of the bone Organ Specific Project (B-OSP) is to address this knowledge gap and generate high quality, single- cell resolution, longitudinal imaging and multi-omics data of normal bones across the human lifespan. We propose the following three specific aims: 1) To refine protocols for biospecimen processing, multi-omics and imaging assays and define inter-individual variability using our existing banked bone samples. Using existing biobanked normal bones from the autopsy program, we will perform a series of pilot studies to refine our protocols for biospecimen collection and processing, as well as single-cell omics and imaging assays. 2) To procure, archive, and annotate high-quality normal bone samples across human life span. We have established a streamlined procurement and biorepository infrastructure to support our bone OSP. We will procure normal bone from the same donor of 6 age groups across the human life span. 3) To spatially profile bone specimens across the human life span using a set of robust and scalable imaging and single-cell omics assays. We will use micro computed tomography to image every bone we collect at both the macroscale and mesoscale. At the microscale, we will perform single-nucleus RNA sequencing (snRNA-Seq) and single-cell assay for transposase-accessible chromatin using sequencing (scATAC-Seq) of two strategic anatomical sites per bone to determine cell type composition, as well as transcriptional and epigenetic signatures. We will perform multiplexed error-robust fluorescence in situ hybridization (MERFISH, for transcriptomics) and Co- Detection by Indexing (CODEX, for proteomics) analyses.

项目摘要 - 骨骼器官特定项目 骨骼是由三种具有不同功能的组织类型制成的高度复杂的器官，包括来自 造血，间质和内皮谱系。限制在一个器官中，与 彼此之间，这三个组织相互依赖于它们的发育和稳态。空间 各种细胞类型的组织及其在骨头中的串扰是我们知识的主要差距。目标 骨骼器官特定项目（B-SOSP）是解决这一知识差距，并产生高质量，单一的 整个人类寿命的正常骨骼的细胞分辨率，纵向成像和多摩学数据。我们 提案以下三个具体目标：1）完善生物环境处理，多摩学和的协议 成像测定并使用我们现有的银行骨样品来定义个体间的变异性。使用现有 尸检计划中的生物群正常骨骼，我们将进行一系列试验研究以完善我们的 生物测量收集和加工的协议，以及单细胞的OMICS和成像测定法。 2）到 在人类寿命中采购，存档和注释高质量的正常骨样品。我们有 建立了简化的采购和生物座基础设施，以支持我们的骨骼OSP。我们将 从整个人类寿命的6岁年龄段的同一捐赠者中采购正常骨骼。 3）到空间剖面 使用一组健壮且可扩展的成像和单细胞OMIC 测定。我们将使用微型计算机断层扫描图像我们在宏观和 中尺度。在微观上，我们将执行单核RNA测序（SNRNA-SEQ）和单细胞 使用两个策略解剖位点的测序（SCATAC-SEQ）进行转座酶可访问的染色质测定 每骨确定细胞类型组成，以及转录和表观遗传特征。我们将 执行多路复用误差荧光原位杂交（用于转录组学）和共同 通过索引（对于蛋白质组学）分析进行索引检测。