Abstract: SA-PO488

Shotgun Cellfree DNA Sequencing to Evaluate Renal Allograft Damage in Recipients with BKVN

Session Information

Category: Transplantation

  • 1702 Transplantation: Clinical and Translational

Authors

  • Dadhania, Darshana, Weill Cornell Medical College, New York, New York, United States
  • Burnham, Philip, Cornell University, Ithaca, New York, United States
  • Lee, John Richard, Weill Cornell Medical College, New York, New York, United States
  • Snopkowski, Catherine, Weill Medical College of Cornell University, New York, New York, United States
  • Li, Carol Y., Weill Cornell Medical College, New York, New York, United States
  • Yang, Hua, Weill Cornell Medical College, New York, New York, United States
  • Muthukumar, Thangamani, Weill Cornell Medical College, New York, New York, United States
  • Suthanthiran, Manikkam, Weill Cornell Medical College, New York, New York, United States
  • De Vlaminck, Iwijn, Cornell University, Ithaca, New York, United States
Background

BKV replication is frequent and is associated with graft loss in 20-50% of cases. Existing non-invasive assays to detect BKV replication fail to correlate with the extent of renal allograft damage. Advances in the measurement of cellfree (cf) DNA in allograft recipients may be useful to identify allograft damage associated with BKVN.

Methods

In 16 recpients with stored urine supernatants, we studied cf DNA of BK virus DNA & donor DNA fractions. Analysis was performed in sex-mismatched individuals with normal protocol biopsy (n=4) & biopsy proven BKVN diagnosis (n=12). We performed shotgun metagenomic sequencing on an Illumina NextSeq (2 x 75 bp) using a single-stranded library preparation. In patients who received organs from the opposite sex, a comparison of the depth of sequencing coverage of the sex chromosomes was used to determine the cf donor DNA fraction and associated with graft injury.

Results

Urine cellfree BKV DNA correlated with urine cell pellet BKV VP1 copies (Fig 1) and Figure 2 demonstrates the correlation between serum creatinine at the time of biopsy and cf donor DNA fraction. Female recipients (n=4) receiving a male donor kidney had significantly lower cf DNA fraction compared to male recipients (n=12) receiving female kidney (Fig 3a). Among the male recipients, those with BKVN diagnosis had significantly higher urine cf donor DNA fraction compared to those with normal protocol biopsy and stable graft function (Fig 3b).

Conclusion

Our data on sex mismatched allograft recipients demonstrates that urinary cf DNA measurement in recipients offers a noninvasive measure of the burden of viral disease & allograft damage. Additional studies are needed to apply this technology to study the dynamic changes in cf donor DNA fraction with increasing/decreasing allograft damage & changes in renal function.