Kidney Week

Abstract: FR-OR052

Precise Editing of the Central Six Nucleotides in the Pkd1-3’UTR miR-17 Binding Motif Is Sufficient to Restore Pkd1 and Slow PKD Progression

Session Information

• Monogenic Kidney Disease: Mechanistic Insights and Therapeutic Approaches
  November 07, 2025 | Location: Room 360A, Convention Center
  Abstract Time: 05:30 PM - 05:40 PM

Category: Genetic Diseases of the Kidneys

• 1201 Genetic Diseases of the Kidneys: Monogenic Kidney Diseases

Authors

• Song, Chunzi, The University of Texas Southwestern Medical Center, Dallas, Texas, United States

Chunzi Song, MD

• Biggers, Laurence Michael, The University of Texas Southwestern Medical Center, Dallas, Texas, United States

Laurence Michael Biggers

• Zumwalt, Maggie, The University of Texas Southwestern Medical Center, Dallas, Texas, United States

Maggie Zumwalt

• Alvarez, Jesus A., The University of Texas Southwestern Medical Center, Dallas, Texas, United States

Jesus A. Alvarez

• Cobo-Stark, Patricia, The University of Texas Southwestern Medical Center, Dallas, Texas, United States

Patricia Cobo-Stark

• Patel, Vishal, The University of Texas Southwestern Medical Center, Dallas, Texas, United States

Vishal Patel, MD

• Lakhia, Ronak, The University of Texas Southwestern Medical Center, Dallas, Texas, United States

Ronak Lakhia, MD

Background

Nearly 80% of ESRD caused by ADPKD is due to mutations in the PKD1 gene. Large deletions encompassing the microRNA-17 binding motif within the Pkd1 3’UTR (P1MBM) can restore Pkd1 and slow cyst growth. Whether a fine-tuned approach to target the central 6 nucleotides in the miR-17 binding motif of the Pkd1 3’UTR will prevent miR-17 hybridization and rescue PKD remains unknown.

Methods

We generated Pkd1^RCΔ6/RC mice using CRISPR/Cas9 in conjunction with a novel donor template, where nucleotides 2-7 of the 8bp P1MBM were mutated. Ksp^Cre;Pkd1^RCΔ6/RC mice were crossed with Pkd1^F/F mice to generate Ksp^Cre;Pkd1^RC/F and Ksp^Cre;Pkd1^RCΔ6/F mice. Tissues were harvested at various time points for histological and molecular analysis, and omics studies. We also designed and tested a small antisense oligonucleotide to sterically bind P1MBM and prevent miR-17 mediated translational repression.

Results

At P18, Ksp^Cre;Pkd1^RCΔ6/F mice exhibited a 75% reduction in kidney-to-body weight ratio, and a 50% decline in serum creatinine compared to Ksp^Cre;Pkd1^RC/F mice. Longitudinal studies with total kidney volume measured by MRI demonstrated a sustained cyst-attenuating effect in Ksp^Cre;Pkd1^RCΔ6/F mice. Western blot (WB) exhibited an increase in PC1, and proteomics identified reduced proliferation and enhanced mitochondrial metabolism in Ksp^Cre;Pkd1^RCΔ6/F mice kidneys. These findings were confirmed by WB and IF staining for pHH3, pCREB, and OXPHOS markers. Interestingly, cMyc, a driver of miR-17, and miR-17 itself remained increased in Ksp^Cre;Pkd1^RCΔ6/F mice, suggesting that the 6bp change is sufficient to decouple the cMyc-miR-17-Pkd1 axis. Hence, we designed a 16-base pair anti-sense oligonucleotide and tested if it could hybridize to the Pkd1 3’UTR to mask P1MBM. Indeed, Pkd1 ASO stabilized Pkd1 mRNA, increased PC1 protein, slowed cyst growth, and improved metabolism in Pkd1^RC/- and human ADPKD kidney cyst derived cells.

Conclusion

Mutation of 6 cardinal nucleotides within the Pkd1 3’UTR is sufficient to rescue a mouse model of ADPKD, independent of miR-17 expression. Steric blockade of the P1MBM with a small ASO similarly stabilized Pkd1 mRNA, increased PC1 protein and reduced cysts in-vitro. These studies define a precise novel target within the Pkd1 3’UTR to treat ADPKD.

Funding

• NIDDK Support

Digital Object Identifier (DOI)

• doi: 10.1681/ASN.20256fcah51a