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Kidney Week

Abstract: TH-PO125

Translating Ribosome Affinity Purification (TRAP) Molecular Profiling of Kidney Cell Types: A Data Resource

Session Information

Category: Acute Kidney Injury

  • 103 AKI: Mechanisms

Authors

  • Huang, Jie, Takeda California, Inc., San Diego, California, United States
  • Vekich, John, Takeda California, Inc., San Diego, California, United States
  • Bobkov, Konstantin, SGI, San Diego, California, United States
  • Holub, Corine, Takeda California, Inc., San Diego, California, United States
  • Das, Debashree, Takeda California, Inc., San Diego, California, United States
  • Taskar, Pranali, NGM Biopharmaceuticals, Santa Clara, California, United States
  • Ekstrand, Mats I., Takeda California, Inc., San Diego, California, United States
  • Yoo, Eun ji, Takeda California, Inc., San Diego, California, United States
  • Chen, Naishan, Takeda California, Inc., San Diego, California, United States
  • Huang, Huey-Jing, Takeda California, Inc., San Diego, California, United States
  • Mcsweeney, Sara J., Clinical Network Services, Bishop?s Stortford, United Kingdom
  • Bedford, David C., Mission Therapeutics Ltd, Cambridge, United Kingdom
  • Doyle, Joseph P., Takeda California, Inc., San Diego, California, United States
Background

The Translating Ribosome Affinity Purification (TRAP) methodology was developed in the labs of Nathaniel Heintz and Paul Greengard at Rockefeller University in order to overcome the inherent cellular complexity of tissues and to obtain cell-type specific translational profiles of critically relevant cell types in these tissues (Doyle et al., Heimann et al., Cell, 2008). Using bacTRAP transgenic mouse lines licensed from Rockefeller University or generated at Takeda, California, we here apply the TRAP methodology to profile relevant kidney cell types, both in a basal state as well as in models of acute kidney injury.

Methods

We employed the TRAP methodology in order to obtain translational profiling data from multiple kidney cell types. Transgenic mice expressing the EGFP-ribosomal L10a fusion protein under kidney cell type specific promoters were generated using standard techniques and characterized for EGFP-L10a expression in the cell types of interest. For collection of TRAP samples, whole kidneys were isolated from several mice, homogenized, and processed following standard TRAP protocols. Total tissue mRNA and cell-type enriched TRAP mRNA samples were amplified, libraries prepared, and sequenced following routine protocols.

Results

We have collected kidney TRAP data covering two general areas: 1. Baseline data, collected from unperturbed transgenic bacTRAP mice, targeting specific cell types (podocytes, proximal tubule epithelial cells (PTECs) and subsets of PTECs, and perivascular mesenchymal cells) 2. Data from disease model studies (sepsis, bilateral ischemia-reperfusion).

Conclusion

Here we demonstrate that bacTRAP is a powerful tool for translational profiling of kidney cell types. In order to make best use of our kidney TRAP datasets, we are making this data available to the greater scientific community. This data is an invaluable resource for identification of potential drug targets and biomarkers for kidney disease, as well as for elucidating the mechanism of action for kidney disorders and drug response. In addition to the RNAseq data, we have access to many TRAP mouse lines on the C57Bl6/J strain which are also available. Details of data and transgenic mouse lines will be available at this poster session.

Funding

  • Commercial Support