Kidney Week

Abstract: TH-OR105

Time Course Analysis of Serial Biopsies Post-Kidney Transplant Reveals a Time Zero Cell State Negatively Associated with Delayed Graft Function

Session Information

• Transplantation: Basic and Translational Advances
  October 24, 2024 | Location: Room 25, Convention Center
  Abstract Time: 05:20 PM - 05:30 PM

Category: Transplantation

• 2101 Transplantation: Basic

Authors

• Malone, Andrew F., St Vincent's University Hospital, Dublin, Ireland

Andrew F. Malone, MD, MBChB, FASN

• Leckie-Harre, Aidan, Washington University in St Louis School of Medicine, St Louis, Missouri, United States

Aidan Leckie-Harre

• Asthana, Amish, Wake Forest University School of Medicine, Winston-Salem, North Carolina, United States

Amish Asthana, PhD

• Paul, Rohan Singh, Washington University in St Louis School of Medicine, St Louis, Missouri, United States

Rohan Singh Paul, MBBS

• Orlando, Giuseppe, Wake Forest University School of Medicine, Winston-Salem, North Carolina, United States

Giuseppe Orlando, MD, PhD

• Humphreys, Benjamin D., Washington University in St Louis School of Medicine, St Louis, Missouri, United States

Benjamin D. Humphreys, MD, PhD, FASN

Background

One fifth of the kidneys procured for transplantation in the US are discarded. Identification of kidneys with sufficient functional reserve at the time of procurement may increase the number of kidneys transplanted. We hypothesized that single-cell RNA-seq analysis of allograft biopsies obtained at time 0, 1 and 12 months will identify a signature of functional reserve predictive of successful graft function in marginal organs and thus minimize discard rates.

Methods

As part of the Double R study (IRB00027118) we analysed time 0, 1 and 12 month biopsies of DCD kidney allografts from 6 patients with either DGF (n=3) or no DGF (n=3). We performed single nucleus RNA-seq on snap frozen biopsies using the 10X FLEX kit. We used Seurat and Monocle2 to generate data objects and perform analyses. We also estimated the proportion of male cells with chromosome Y loss (LOY) per cell type over time.

Results

We integrated 17318 post-QC cells from 6 patients with and without DGF across three time points (0, 1, 12 months) post DCD kidney transplant. These included 5658 proximal tubule cells. Using Monocle we identified a cell state present in time 0 biopsies that did not develop DGF (figure1A+B). This cell state was absent in biopsies that progressed to DGF but recovered by 1 month. This cell state was defined by upregulation of fructose metabolism genes and had a high median LOY proportion (0.2459) (figure1C).

Conclusion

We identified a time zero cell state that is negatively associated with DGF in the setting of DCD kidney transplantation. These renal proximal tubule cells differentially express fructose metabolism genes and have a high proportion of LOY.

Figure1. Monocle pseudotime analysis of proximal tubule cells defines 9 cell states (A+B). Cell state 5 is absent at time 0 in biopsies that progress to DGF (B). This cell state has a high proportion of chromosome Y loss (C).