Abstract: FR-PO593
mRNA Translation Reprogramming in Autosomal Dominant Polycystic Kidney Disease (ADPKD)
Session Information
- Cystic Kidney Diseases: Mechanisms and Models
October 25, 2024 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Genetic Diseases of the Kidneys
- 1201 Genetic Diseases of the Kidneys: Cystic
Authors
- Leemans, Charlotte, Centre Hospitalier Universitaire de Liege, Liege, Belgium
- El Hachem, Najla, Universite de Liege, Liege, Belgium
- Decuypere, Jean-Paul, Katholieke Universiteit Leuven Universitaire Ziekenhuizen Leuven Campus Gasthuisberg, Leuven, Flanders, Belgium
- Mekahli, Djalila, Katholieke Universiteit Leuven Universitaire Ziekenhuizen Leuven Campus Gasthuisberg, Leuven, Flanders, Belgium
- Jouret, Francois, Centre Hospitalier Universitaire de Liege, Liege, Belgium
- Close, Pierre, Universite de Liege, Liege, Belgium
Group or Team Name
- Laboratory of Cancer Signaling, Laboratory of Translational Research in Nephrology.
Background
Global regulation of mRNA translation has emerged as a central mechanism driving cellular adaptation in various diseases. tRNA modifications play a specific role in promoting the reprogramming of mRNA translation. We aim to better understand the importance of mRNA translation reprogramming in the development of cysts in ADPKD.
Methods
Murine inner medullary collecting duct (IMCD) cells knocked out for Pkd1 by CRISPR-Cas9 were grown in vitro. A range of different techniques were used to characterize mRNA translation regulation in these cells. First, we compared the proteome of control or Pkd1 KO cells by label-free quantitative proteomics. In parallel, we assessed mRNA translation regulation by polysome profiling and OP-Puro incorporation. Finally, we purified tRNAs from cells and we performed a systematic analysis of most of the described tRNA modifications by LC-MS/MS analysis.
Results
We generated lists of proteins that are up- and down-regulated upon loss of Pkd1 in IMCD cells. We focused on proteins whose expression is upregulated in Pkd1 KO as compared to controls. Performing Gene Set Enrichment Analyses, we uncovered a series of specific cellular and molecular pathways that are significantly upregulated in Pkd1 KO cells. We found that “tRNA modification” was enriched in cells KO for Pkd1: a series of enzymes, such as Dus3l or Pus10, associated with tRNA modification were specifically upregulated. We plotted enzymes known to regulate tRNA modification and compared their expression in control versus Pkd1 KO cells. These data were corroborated with a systematic analysis of tRNA modifications. Interestingly, this approach highlighted reproducible changes in specific tRNA modifications between control and Pkd1 KO cells. Differences in abundance of each tRNA modification were plotted and correlated with catalyzing enzyme expression.
Conclusion
Our data indicate that differences in tRNA modification pattern are observed between control and Pkd1 KO IMCD cells and could define specific tRNA modification pathways as a new vulnerability targets in cyst formation in ADPKD.
Funding
- Government Support – Non-U.S.