Washington University Kidney O’Brien Center for Chronic Kidney Disease Research
Summary
The Washington University Kidney O’Brien Center for Chronic Kidney Disease Research (WUCKD-NRC) primary goals are to develop and disseminate tools and new technologies for the investigation of chronic kidney disease and fibrosis. The WUCKD-NRC achieves these goals through two Biomedical Cores – the Metabolism Core (Meta-Core) and the Variant Validation Core.
Metabolism Core (Meta-Core)
-
Description
The aim of the Metabolism Core (Meta-Core) is to facilitate the use of metabolic assays in pre-clinical models of chronic kidney disease (CKD) for the kidney community by providing the following services following consultation with core faculty:
-
Services
- Seahorse bioflux analyzer (XF24/XF96)
- Measures metabolism and define metabolic substrate preferences ideally on adherent cells but also on freshly isolated tubules.
- High resolution respirometry (Oroboros)
- Measures mitochondrial function in tissues ex vivo, and this is facilitated through the Nutrition Obesity Research Center (NORC).
- Substrate oxidation assays
- Utilizes radioactive-labeled substrates (e.g. 3H-palmitate) to measure oxidation in tissue ex vivo.
- Untargeted metabolomics
- Performed on cells or tissues using the mass spectrometry resources of the Metabolomics Core through the lab of Gary Patti.
- Stable isotope flux studies
- Used to look at specific pathway activity and flux in both cell culture systems and whole animals using mass spectrometry.
- Seahorse bioflux analyzer (XF24/XF96)
Variant Validation Core
-
Description
The widespread and increasing use of whole exome sequencing (WES) and whole genome sequencing (WGS) to investigate the genetic bases for CKD and other kidney disorders has led to a much better understanding of the etiology of monogenic diseases. The aim of the Variant Validation Core (VVC) is to investigate variants of uncertain significance (VUS) discovered in patients with kidney disease or a kidney developmental disorder for potential pathogenicity. The VVC will use experimental approaches in cultured cells and in mice. The experimental approach for each VUS will be tailored for 1) the particular gene and protein that are potentially involved; and 2) the particular disease or disorder that would likely develop if the VUS is indeed pathogenic. Immediate services and resources that are being offered include:
-
Services
- COL4A3/A4/A5 VUS (missense) analysis by split nano-Luciferase collagen IV assembly and secretion assays using transfected cells
- COL4A3/A4/A5 VUS (intronic) analysis by RNA splicing assays using transfected cells
- COL4A3/A4/A5 VUS analysis by high resolution quantitative immunofluorescence imaging in paraffin sections of kidney biopsies
- COL4A3/A4/A5 VUS analysis in vivo by mouse gene editing
- Analysis of VUS in other genes through design and implementation of tailored laboratory assays
Contact Information
If you would like more information regarding services provided by WUCKD-NRC, please contact:
Benjamin Humphreys
Website