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Abstract: PO1707

Glomerular mRNAs Are Alternatively Spliced and Polyadenylated During Podocyte Injury in Animal Models of Nephrotic Syndrome

Session Information

Category: Glomerular Diseases

  • 1204 Podocyte Biology

Authors

  • Burton, Claire, Abigail Wexner Research Institute at Nationwide Children's Hospital, Columbus, Ohio, United States
  • Webb, Amy, The Ohio State University, Columbus, Ohio, United States
  • Cianciolo, Rachel, The Ohio State University, Columbus, Ohio, United States
  • Moore, Claire L., Tufts University School of Medicine, Boston, Massachusetts, United States
  • Agrawal, Shipra, Abigail Wexner Research Institute at Nationwide Children's Hospital, Columbus, Ohio, United States
Background

Glomerular disease, often manifesting as nephrotic syndrome (NS) with high proteinuria, is characterized by podocyte loss and injury. Furthermore, alternative mRNA processing, such as alternative splicing (AS) and alternative polyadenylation (APA) play important roles in physiology, development, and disease; however, there is very limited knowledge of their roles in glomerular disease. We hypothesized that AS and APA events of glomerular RNAs is associated with podocyte injury and proteinuria in NS.

Methods

Glomerular damage characterized by proteinuria was induced by puromycin aminonucleoside (PAN) or adriamycin (ADR) to mimic human minimal change disease (MCD) or focal segmental glomerulosclerosis (FSGS), respectively. Urine and serum chemistries, kidney histology and glomerular RNA-seq analyses were performed. APATrap and JunctionSeq bioinformatics analyses software were used to detect APA and AS glomerular events. Correlation of differentially expressed genes (DEGs) was performed with known glomerular disease genes and polyadenylation and splicing factors.

Results

Robust proteinuria was induced in both PAN-MCD and ADR-FSGS models, accompanied by hypoalbuminemia, hypercholesterolemia and histological alterations in the kidneys (protein casts and podocyte hypertrophy). Out of 13,265 genes, MCD model resulted in 1033 and FSGS model in 1308 glomerular DEGs with abs(log2FC)>1 and Padj<0.05. Of 80 analyzed genes with established roles in glomerular disease, 30 were altered in both MCD and FSGS. Significant APA was identified in 71 and 746 genes in MCD and FSGS nephrosis, respectively, and of 173 polyadenylation factors analyzed, 21 were altered in MCD and 24 in FSGS. Significant AS was identified in 136 and 1875 genes in MCD and FSGS models, respectively. In accordance, of 50 splicing factors analyzed, 3 were altered in MCD and 5 in FSGS. Specifically, the identified APA and AS events affected genes of the slit diaphragm complex such as Nphs1, Nphs2, and Tjp1, which are critical determinants of podocyte structure and function.

Conclusion

Association of global glomerular mRNA alteration due to AS and APA with podocyte and glomerular injury is a newly recognized phenomenon, with potential implications for therapy and molecular understanding of the disease.

Funding

  • Other NIH Support