Kidney Week

Abstract: TH-PO735

Fabry Nephropathy: Transcriptome Sequencing of Microdissected Renal Compartments from Archival Kidney Biopsies at Baseline, and After 5 and 10 Years of Enzyme Replacement Therapy

Session Information

• Genetic Diseases of the Kidneys: Non-Cystic - I
  October 25, 2018 | Location: Exhibit Hall, San Diego Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Genetic Diseases of the Kidney

• 1002 Genetic Diseases of the Kidney: Non-Cystic

Authors

• Eikrem, Oystein, University of Bergen, Bergen, Norway

Oystein Eikrem,

• Strauss, Philipp, University of Bergen, Bergen, Norway

Philipp Strauss,

• Sekulic, Miroslav, Brigham and Women's Hospital , Boston, Massachusetts, United States

Miroslav Sekulic,

• Tøndel, Camilla, Haukeland University Hospital, Bergen, Norway

Camilla Tøndel,

• Flatberg, Arnar, Norwegian University of Science and Technology, Trondheim, Norway

Arnar Flatberg,

• Skrunes, Rannveig, Haukeland University Hospital, Bergen, Norway

Rannveig Skrunes,

• Landolt, Lea, University of Bergen, Bergen, Norway

Lea Landolt,

• Babickova, Janka, University of Bergen, Bergen, Norway

Janka Babickova,

• Leh, Sabine, Haukeland University Hospital, Bergen, Norway

Sabine Leh,

• Scherer, Andreas, Spheromics, Kontiolahti, Finland

Andreas Scherer,

• Svarstad, Einar, Haukeland University Hospital, Bergen, Norway

Einar Svarstad,

• Marti, Hans-Peter, University of Bergen, Bergen, Norway

Hans-Peter Marti,

Background

The present study exploited next generation mRNA sequencing from kidney biopsies of Fabry patients with a classical phenotype

Methods

Kidney biopsies (n=15) were obtained with a 16G needle from Fabry patients (n=5) from 2003-2014. Median patient age at baseline biopsy was 18 years (7-30 y). Control biopsies from peri-tumorous tissue of renal cancer patients (n=32). Formalin-fixed, paraffin-embedded (FFPE) kidney biopsies were obtained: a) baseline prior to enzyme replacement therapy (ERT; n=5), b) after 5y of ERT (n=5), and c) 8-10y of ERT (n=5). Total RNA was extracted from laser-captured, microdissected glomerular, proximal tubular, distal tubular and arterial cross-sections (High Pure FFPET RNA extraction kit; Roche). cDNA libraries were prepared using the TruSeq RNA Access Library Prep Kit^® (Illumina) and sequenced on an Illumina HiSeq 2500/HiSeq 4000 instrument

Results

We present the results of the first 42 samples of a larger ongoing series. Several hundred mRNAs were differentially expressed (p-value <0.05, abs. fold change >2) between normal controls and Fabry baseline samples, and between the baseline samples and the two different treatment time points. The samples segregated completely within their own compartment (Fig. 1) in principle component analysis. Gene Set Enrichment Analysis in the glomerular compartment demonstrated enriched gene sets of extracellular matrix, EMT, fibrosis, and immune response in the 10 year biopsies compared to baseline (p<0.05) despite ERT. Several key regulatory inflammatory genes were found to be upregulated in all compartments

Conclusion

NGS is feasible in archival Fabry disease kidney biopsies, expanding the potential utility of kidney biopsies to detect markers of Fabry nephropathy, chronic kidney disease and therapeutic targets

Funding

• Commercial Support –