Abstract: FR-PO968
Integration of Spatial Metabolomics to Single-Nucleus Droplet-Based Sequencing Data Identifies New Glomerulus-Specific Gene Markers
Session Information
- Pathology and Lab Medicine: Basic
November 08, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Pathology and Lab Medicine
- 1601 Pathology and Lab Medicine: Basic
Authors
- Zhang, Guanshi, University of Texas Health San Antonio, San Antonio, Texas, United States
- Velickovic, Dusan, Pacific Northwest National Laboratory, Richland, Washington, United States
- Pamreddy, Annapurna, University of Texas Health San Antonio, San Antonio, Texas, United States
- Lake, Blue, University of California San Diego, La Jolla, California, United States
- Drel, Viktor, University of Texas Health San Antonio, San Antonio, Texas, United States
- Kaushik, Dharam, University of Texas Health San Antonio, San Antonio, Texas, United States
- Bansal, Shweta, University of Texas Health San Antonio, San Antonio, Texas, United States
- Venkatachalam, Manjeri A., University of Texas Health San Antonio, San Antonio, Texas, United States
- Pasa-Tolic, Ljiljana, Pacific Northwest National Laboratory, Richland, Washington, United States
- Alexandrov, Theodore, European Molecular Biology Laboratory, Heidelberg, Germany
- Anderton, Christopher R., Pacific Northwest National Laboratory, Richland, Washington, United States
- Sharma, Kumar, University of Texas Health San Antonio, San Antonio, Texas, United States
Group or Team Name
- for the KPMP Consortium
Background
The current study is a part of the NIDDK Kidney Precision Medicine Project (KPMP). We recently cross-validated that a sphingomyelin (SM(d18:1/16:0)) was a glomerulus-enriched marker, as visualized by two independent sites using matrix-assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI). To establish integration of data with other Tissue Interrogation Sites (TIS) and technologies, we provided a tabulated list of 48 SM/ceramide metabolism-related gene/enzymes (from KEGG Ontology Database) to results from other TIS sites.
Methods
Two MALDI-MSI platforms (QE-HFX at UTHSA and FTICR at PNNL) were employed to spatially characterize the lipid profile in normal human kidney tissues (n = 6; U. Michigan) at 20-30 μm spatial resolution. 30 different cell types were identified using the single-nucleus droplet-based sequencing (snDrop-Seq) analysis developed by the UCSD-WashU TIS and the SM/ceramide genes were queried across the database. Enrichr software and the Database of Genotypes and Phenotypes (dbGaP) were used for enrichment analysis.
Results
PLPP3was identified as one of the few glomerular endothelial cell specific genes and can be added as a gene marker to KPMP Atlas. Furthermore, PLPP1was found to be localized to the glomerular endothelial cells and to endothelial cells of AEA and descending vasa recta. Fluorescence microscopy analysis showed that PLPP3 protein is localized within glomeruli but separated from synaptopodin (a podocyte marker). Further enrichment analysis using Enrichr and dbGaP showed that the identified glomerular specific genes are associated with carotid stenosis, diabetic nephropathies, and other phenotypes.
Conclusion
This highlights the value of the untargeted mass spectrometry imaging for linking the spatial metabolomic data to different kidney cells types, and an avenue connect metabolic information to gene expression. Integrated MALDI-MSI and snDrop-Seq data could help identify novel glomerular-specific gene markers.
Funding
- NIDDK Support