Vanderbilt University Biomolecular Multimodal Imaging Center for 3-Dimensional Mapping of the Human Kidney

项目摘要-肾脏OSP。 人类肾脏内不同细胞类型的整体结构和解剖关系良好。 理解，实验研究已经定义了个体的分子和功能特征 肾细胞类型。然而，值得注意的是，人们对整合、相互作用和分子交叉知之甚少。 正常肾脏中不同的细胞室之间的谈话。肾脏OSP的目标是解决 通过开发一种完全集成的、多模式的分子成像管道来解决当前知识中的这一局限性 描述人类肾脏的细胞和分子组织。在我们已经建立的 HuBMAP表征流水线，我们将把成像质谱学汇聚在一起，高度复用 免疫荧光显微镜，自发荧光显微镜，染色显微镜，空间转录学， 空间蛋白质组学和单细胞RNA序列。每一种模式都是专门选择的，以使构建成为可能 肾脏分子图谱跨越广泛的空间尺度(例如，单细胞，功能组织单元， 邻居和整个器官)和分子类别(例如代谢物、脂类、蛋白质和RNA)。这个 结果将是一个全面的人类肾脏的分子图谱，它将与其他 分子讯问管道和HuBMAP研究人员生成的数据。在目标1中，我们将继续我们的 以我们已建立的肾脏生物库和组织为基础的肾组织收集和管理方案 收购计划，以创建具有代表性的人类肾脏分子图谱。目标2建立临床 以及组织学标准和衡量标准，将用于确定样本的适合性 进入了分析管道。目标3的焦点提出了2-D和3-D多模式成像的策略 对人类肾脏进行深入的多组学分析。手术切除标本的分析进展将是 应用于使用不可移植的供体肾脏构建三维分子图谱的整个器官。最后，瞄准 4将扩大肾脏特异性表征管道，以产生具有代表性的3-D分子图谱 来自多个完整的人类肾脏。

PROJECT SUMMARY – Kidney OSP. The overall structure and anatomical relationships among different cell types within the human kidney are well understood, and experimental studies have defined the molecular and functional characteristics of individual kidney cell types. However, remarkably little is known about the integration, interactions, and molecular cross- talk between the different cellular compartments in normal kidneys. The goal of the Kidney OSP is to address this limitation in current knowledge by developing a fully integrated, multimodal molecular imaging pipeline to characterize the cellular and molecular organization of the human kidney. Building on our already established HuBMAP characterization pipeline, we will bring together imaging mass spectrometry, highly multiplexed immunofluorescence microscopy, autofluorescence microscopy, stained microscopy, spatial transcriptomics, spatial proteomics, and single-cell RNA-seq. Each modality was specifically chosen to enable the construction of a kidney molecular atlas that spans a wide range of spatial scales (e.g., single cells, functional tissue units, neighborhoods, and whole organs) and molecular classes (e.g., metabolites, lipids, proteins, and RNAs). The result will be a comprehensive molecular atlas of the human kidney that will be fully compatible with other molecular interrogation pipelines and data generated by HuBMAP researchers. In Aim 1 we will continue our kidney tissue collection and management protocols that build on our established kidney biobanks and tissue acquisition programs to create a representative molecular atlas of the human kidney. Aim 2 establishes clinical and histological standards and metrics that will be used to determine the suitability of samples that can be entered into the analytical pipeline. The focus of Aim 3 puts forth a strategy for 2-D and 3-D multimodal imaging and deep multi-omic analysis of the human kidney. Assay development on surgical resection specimens will be applied to whole organs using non-transplantable donor kidneys to construct 3-D molecular atlases. Finally, Aim 4 will scale up the kidney-specific characterization pipeline to generate representative 3-D molecular atlases from multiple, intact human kidneys.