Kidney Week

Abstract: TH-PO601

Anks3 Knockout in Rats Causes Prenatal Lethality Due to Major Disturbances in Organ Patterning

Session Information

• Cystic Kidney Diseases - I
  November 02, 2017 | Location: Hall H, Morial Convention Center
  Abstract Time: 10:00 AM - 10:00 AM

Category: Genetic Diseases of the Kidney

• 801 Cystic Kidney Diseases

Authors

• Clark, Euan, University of Heidelberg, Mannheim, Germany

Euan Clark,

• Papagiannarou, Stamatia Matina, University of Heidelberg, Mannheim, Germany

Stamatia Matina Papagiannarou,

• Gosmann, Christian, University of Heidelberg, Mannheim, Germany

Christian Gosmann,

• Groene, Hermann-Josef, German Cancer Research Center, Heidelberg, Germany

Hermann-Josef Groene,

• Dobreva, Gergana D, University of Heidelberg, Mannheim, Germany

Gergana D Dobreva,

• Lelongt, Brigitte, UMR_S1155, PARIS, France

Brigitte Lelongt,

• Gauguier, Dominique, INSERM, Paris, France

Dominique Gauguier,

• Bowie, James, University of California, Los Angeles, Los Angeles, California, United States

James Bowie,

• Hoffmann, Sigrid C., University of Heidelberg, Mannheim, Germany

Sigrid C. Hoffmann,

Background

Anks3 was first discovered as a novel interacting partner of Anks6. The Anks6^R823W mutation disrupts binding to Anks3 and causes dominant polycystic kidney disease (ADPKD) in rats. Though its function remains obscure, Anks3 involvement in nephronophthisis was suggested. The present study aimed to study the consequences of a total Anks3 knockout (Anks3KO) in rats.

Methods

Anks3KO rats were generated by injection of a gRNA targeting the second exon of Anks3 (aa5-12) and Cas9 mRNA into fertilized Sprague Dawley rat zygotes. Rats were genotyped by sequencing the amplified targeted region. RT-PCR and Western blotting were used to prove loss of Anks3. We determines clinical parameters in 24-hr urine and blood plasma to evaluate renal function Histological analysis was performed by standard techniques.

Results

Founders were obtained carrying either a 5bp or an 8bp deletion at the targeted position, which resulted in a premature translational stop at positions aa46 and aa45 respectively, and were used for breeding. Among 82 progenies, 39% were genotyped as wildtype and 61% as heterozygotes. Homozygous Anks3KO rats frequently died in mid-gestation and never survived birth. They exhibited a complex pattern of disturbances in organ patterning including defects in cortex and midbrain development, craniofacial defects, focal hemorrhages and complex structural heart defects. Kidney size was often recduced and associated with abnormal glomerulogenesis and tubulogenesis. Heterozygous Anks3 +/- rats survived and developed mild kidney abnormalities when aged, including small medullary cysts and fibrosis in the cortico-medullary renal border. Urine production was significantly less in Anks3 +/- rats compared to WT littermates. However, Anks3KO rats never developed an ADPKD-like phenotype, as seen in Anks6^R823W rats.

Conclusion

Conclusion: Here we provide the first evidence that Anks3KO in mammals causes major, lethal developmental disturbances in organ patterning, similarly to heterotaxy mutants with cilia defects, and a nephronophthisis like phenotype in heterozygous adults. We conclude that Anks3 might be a major factor in regulating ciliary function and controls urine concentration. This study was supported by the grant of the NIH (5RO1DK100482) to JB and SH

Funding

• NIDDK Support