PROJECT SUMMARY – Overall This application proposes the continuation of a Tissue Mapping Center (TMC) for the human kidney within the Human BioMolecular Atlas Program (HuBMAP). The mission of the proposed TMC is to build a platform of integrated technologies for imaging and molecular analysis that enables the construction of comprehensive 3- dimensional molecular atlases of the human kidney. This TMC will leverage the unique resources of the Mass Spectrometry Research Center and the National Research Resource for Imaging Mass Spectrometry at Vanderbilt University, the world-class clinical environment of the Vanderbilt University Medical Center, and the advanced biocomputational infrastructure available to the TMC through the Data Analysis Core laboratories at Vanderbilt University and the Delft University of Technology, Netherlands to create a capability to molecularly characterize human tissues in 3-dimensions at a level of understanding unrivaled by current technologies. The innovative aspect of the proposed TMC is the integration of imaging mass spectrometry, highly multiplexed immunofluorescence microscopy, autofluorescence microscopy, stained microscopy, spatial transcriptomics, spatial proteomics, and single-cell RNA-seq to create comprehensive 2-D and 3-D molecular and cellular atlases of the human kidney. Through segmentation and cell type analysis determined by classical markers, we will correlate our molecular and clinical information to the existing knowledge of the kidney to better define the normal phenotypes across a range of diverse samples. The application of this multimodal pipeline to the kidney will provide a new paradigm of understanding the normal state of this organ across multiple dimensions, both molecular (e.g., lipids, metabolites, proteins, and transcripts) and spatial (e.g., whole organs to single cells). As a HuBMAP participant, the molecular atlases produced by this TMC will be disseminated to the community and to collaborators to generate new hypotheses regarding the function of this important organ system, enabling new insight into human health and disease. This multidisciplinary effort requires the creation and integration of three capabilities to 1) procure and manage human tissue specimens, 2) determine and mitigate non-biological pre- analytical factors, and 3) acquire, process, and disseminate multimodal 3-dimensional imaging and large-scale omics data. The goal of the proposed kidney TMC is to create atlases for community users, physicians, and researchers to navigate across the anatomical and molecular landscape of the kidney, generate novel hypotheses, and tackle meaningful biomedical research questions.