Related Experiment Video
Updated: Aug 9, 2025

Implementing Patch Clamp and Live Fluorescence Microscopy to Monitor Functional Properties of Freshly Isolated PKD Epithelium
Published on: September 1, 2015
Functional TFEB activation characterizes multiple models of renal cystic disease and loss of polycystin-1
Jonathan M Shillingford1, James A Shayman1
1Department of Internal Medicine, University of Michigan Medical School, University of Michigan, Ann Arbor, Michigan, United States.
Abstract:
Polycystic kidney disease is a disorder of renal epithelial growth and differentiation. Transcription factor EB (TFEB), a master regulator of lysosome biogenesis and function, was studied for a potential role in this disorder. Nuclear translocation and functional responses to TFEB activation were studied in three murine models of renal cystic disease, including knockouts of folliculin, folliculin interacting proteins 1 and 2, and polycystin-1 (Pkd1) as well as in mouse embryonic fibroblasts lacking Pkd1 and three-dimensional cultures of Madin-Darby canine kidney cells. Nuclear translocation of Tfeb characterized cystic but not noncystic renal tubular epithelia in all three murine models as both an early and sustained response to cyst formation. Epithelia expressed elevated levels of Tfeb-dependent gene products, including cathepsin B and glycoprotein nonmetastatic melanoma protein B. Nuclear Tfeb translocation was observed in mouse embryonic fibroblasts lacking Pkd1 but not wild-type fibroblasts. Pkd1 knockout fibroblasts were characterized by increased Tfeb-dependent transcripts, lysosomal biogenesis and repositioning, and increased autophagy. The growth of Madin-Darby canine kidney cell cysts was markedly increased following exposure to the TFEB agonist compound C1, and nuclear Tfeb translocation was observed in response to both forskolin and compound C1 treatment. Nuclear TFEB also characterized cystic epithelia but not noncystic tubular epithelia in human patients with autosomal dominant polycystic kidney disease. Noncanonical activation of TFEB is characteristic of cystic epithelia in multiple models of renal cystic disease including those associated with loss of Pkd1. Nuclear TFEB translocation is functionally active in these models and may be a component of a general pathway contributing to cystogenesis and growth.NEW & NOTEWORTHY Changes in epithelial cell metabolism are important in renal cyst development. The role of TFEB, a transcriptional regulator of lysosomal function, was explored in several models of renal cystic disease and human ADPKD tissue sections. Nuclear TFEB translocation was uniformly observed in cystic epithelia in each model of renal cystic disease examined. TFEB translocation was functionally active and associated with lysosomal biogenesis and perinuclear repositioning, increased TFEB-associated protein expression, and activation of autophagic flux. Compound C1, a TFEB agonist, promoted cyst growth in 3-D cultures of MDCK cells. Nuclear TFEB translocation is an underappreciated signaling pathway for cystogenesis that may represent a new paradigm for cystic kidney disease.
Insights
Transcription factor EB (TFEB) activation is a key feature in polycystic kidney disease, driving cyst growth. This finding in multiple models suggests a new therapeutic target for renal cystic diseases.
Area of Science:
- Nephrology
- Cell Biology
- Molecular Biology
Background:
- Polycystic kidney disease (PKD) involves abnormal renal epithelial cell growth and differentiation.
- Transcription factor EB (TFEB) regulates lysosome biogenesis and cellular waste removal.
- The role of TFEB in PKD pathogenesis was previously unclear.
Purpose of the Study:
- To investigate the role of TFEB in the development and progression of renal cystic diseases.
- To determine if TFEB activation is a common feature across different PKD models.
- To explore TFEB's functional impact on cyst formation and growth.
Main Methods:
- Studied TFEB nuclear translocation and function in murine models of PKD (folliculin, polycystin-1 knockouts).
- Utilized mouse embryonic fibroblasts lacking Pkd1 and 3D Madin-Darby canine kidney cell cultures.
- Examined TFEB-dependent gene expression, lysosomal activity, and autophagy in cystic epithelia and fibroblasts.
- Assessed TFEB activation in human autosomal dominant polycystic kidney disease (ADPKD) tissue.
Main Results:
- Nuclear TFEB translocation was consistently observed in cystic epithelia across all PKD models and human ADPKD samples.
- TFEB activation correlated with increased expression of TFEB-dependent genes (e.g., cathepsin B).
- Pkd1-deficient cells showed enhanced lysosomal biogenesis, autophagy, and TFEB activity.
- TFEB agonist (Compound C1) significantly increased cyst growth in cell cultures.
Conclusions:
- Noncanonical TFEB activation is a hallmark of cystic epithelia in diverse renal cystic disease models, including Pkd1 loss.
- Nuclear TFEB translocation is functionally active, contributing to cystogenesis and growth.
- TFEB represents a potential therapeutic target and a novel paradigm for treating cystic kidney diseases.
Related Concept Videos
Nephrons
Renal Tubule and Collecting Duct
Proximal Convoluted Tubule (PCT):
The PCT is the initial segment of the renal tubule, extending from the Bowman's capsule that encloses the glomerulus. Its convoluted structure and microvilli-lined cells increase the surface area for reabsorption. The PCT reabsorbs glucose, amino acids, sodium, and water from the filtrate, ensuring essential...

