Organ Specific Project - Bone Marrow

项目总结--骨骼器官专用项目 骨骼是一个高度复杂的器官，由三种不同功能的组织类型组成，包括来自 造血系、间充质系和内皮系。被限制在一个器官内，并与 这三种组织的发育和动态平衡是相互依赖的。空间上的 骨骼中各种细胞类型的组织及其相互作用是我们知识的一个主要缺口。目标是 骨器官特异性项目(B-OSP)的目的是解决这一知识差距，并产生高质量的、单一的 人体正常骨骼的细胞分辨率、纵向成像和多组学数据。我们 提出以下三个具体目标：1)完善生物样品加工、多组学和 使用我们现有的库骨样本进行成像分析并定义个体间的可变性。使用现有的 从尸检程序中提取的正常骨骼生物库，我们将进行一系列初步研究，以完善我们的 生物样品采集和处理方案，以及单细胞组学和成像分析。2)至 获取、存档和注释人类一生中的高质量正常骨样本。我们有 建立了简化的采购和生物信息库基础设施，以支持我们的OSP。我们会 取自6个年龄段的相同供体的正常骨。3)在空间上配置文件 使用一套强大的、可扩展的成像和单细胞组学，在人类生命周期内提供骨标本 化验。我们将使用微型计算机断层扫描技术对我们收集的每一块骨骼进行宏观和微观的成像 中尺度。在微观上，我们将进行单核RNA测序(SnRNA-Seq)和单细胞 利用两个关键解剖部位的测序(scATAC-Seq)检测转座酶可及染色质 以确定细胞类型组成以及转录和表观遗传特征。我们会 执行多重错误稳健荧光原位杂交(MerFish，用于转录)和Co- 通过索引(Codex，用于蛋白质组学)分析进行检测。

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.