Abstract: FR-PO256
Single Gene Mutations in Pkd1 or Tsc2 Alter Extracellular Vesicles Production and Trafficking
Session Information
- Genetic Diseases of the Kidneys: Cystic - II
November 04, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
Abstract Time: 10:00 AM - 12:00 PM
Category: Genetic Diseases of the Kidneys
- 1101 Genetic Diseases of the Kidneys: Cystic
Authors
- Bissler, John J., St. Jude Children's Research Hospital, Memphis, Tennessee, United States
- Kumar, Prashant, US FDA National Center for Toxicological Research, Little Rock, Arkansas, United States
- Al-Zadjali, Fahad, Sultan Qaboos University, Muscat, Oman
- Yao, Ying, The University of Tennessee Health Science Center College of Medicine, Memphis, Tennessee, United States
- Hofherr, Alexis, Albert-Ludwigs-Universitat Freiburg, Freiburg im Breisgau, Baden-Württemberg, Germany
- Kottgen, Michael, Albert-Ludwigs-Universitat Freiburg, Freiburg im Breisgau, Germany
- Mehta, Darshan, US FDA National Center for Toxicological Research, Little Rock, Arkansas, United States
- Gross, Kenneth W., Roswell Park Comprehensive Cancer Center, Buffalo, New York, United States
Background
Patients with autosomal dominant polycystic kidney disease (ADPKD) and the cystic disease associated with tuberous sclerosis complex (TSC) are born with normal or near normal kidneys that later develop cysts and prematurely lose function. Both renal cystic diseases appear to be mediated, at least in part, by disease promoting extracellular vesicles (EVs) that can induce genetically intact cells to participate in the renal disease process.
Methods
We used centrifugation and size exclusion chomotography to isolate the EVs for study. We characterized the EVs using tunable resistive pulse sensing, dynamic light scattering, transmission electron microscopy and western blot analysis. We performed EV trafficking studies using a dye approach in both tissue culture and in vivo studies.
Results
We have previously reported that loss of the Tsc2 gene significantly increased EV production and here demonstrate that the loss of the Pkd1 gene also significantly increases EV production. Using a cell culture system, we also show that loss of either the Tsc2 or Pkd1 gene results in EVs that exhibit an enhanced uptake by renal epithelial cells and a prolonged half-life, possibly further accentuating the EV dose effect. Loss of the primary cilia significantly reduces EV production in renal collecting duct cells. We document that EVs from cells that have loss of polycystin-1 have greatly altered kinetics and a profound effect on the EV half-life, possibly impacting the duration of EV cargo effect on the recipient cell.
Conclusion
These results demonstrate the interplay between primary cilia and EVs and support a role for EVs in polycystic kidney disease pathogenesis.
Funding
- Private Foundation Support