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

项目总结-肾脏OSP。 人体肾脏的整体结构和不同细胞类型之间的解剖关系是良好的 实验研究已经确定了个体的分子和功能特征， 肾细胞类型然而，对整合、相互作用和分子交叉知之甚少。 正常肾脏中不同细胞间的交流。肾脏OSP的目标是解决 通过开发完全集成的多模态分子成像管道， 表征人类肾脏的细胞和分子组织。基于我们已经建立的 HuBMAP表征管道，我们将汇集成像质谱，高度复用 免疫荧光显微术，自体荧光显微术，染色显微术，空间转录组学， 空间蛋白质组学和单细胞RNA-seq。每种模式都是专门选择的， 跨越宽范围的空间尺度的肾分子图谱（例如，单细胞，功能组织单位， 邻域和整个器官）和分子类别（例如，代谢物、脂质、蛋白质和RNA）。的 结果将是一个全面的人类肾脏的分子图谱，将完全兼容其他 分子询问管道和HuBMAP研究人员生成的数据。在目标1中，我们将继续 建立在我们已建立的肾脏生物库和组织基础上的肾脏组织收集和管理方案 采集程序来创建人类肾脏的代表性分子图谱。目标2建立临床 和组织学标准和指标，将用于确定样本的适用性， 进入分析管道。目标3的重点提出了一种用于2-D和3-D多模态成像的策略 以及对人类肾脏的深度多组学分析。手术切除标本的检测开发将 使用不可移植的供体肾脏应用于整个器官以构建3D分子图谱。最后，Aim 4将扩大肾脏特异性表征管道，以生成代表性的3-D分子图谱 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.