Kidney Week

Abstract: FR-PO999

Molecular Signatures of Reperfusion in Ischemic Donor Kidneys: Distinguishing Rejection and Nonrejection Risk Genotypes through Single Nuclear RNA Sequencing

Session Information

• Transplantation: Basic
  October 25, 2024 | Location: Exhibit Hall, Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Transplantation

• 2101 Transplantation: Basic

Authors

• Kuscu, Cem, The University of Tennessee Health Science Center, Memphis, Tennessee, United States

Cem Kuscu, PhD

• Dogan, Murat, The University of Tennessee Health Science Center, Memphis, Tennessee, United States

Murat Dogan, MD, PhD

• Kuscu, Canan, The University of Tennessee Health Science Center, Memphis, Tennessee, United States

Canan Kuscu, PhD

• Randall, Henry B., Saint Louis University, St Louis, Missouri, United States

Henry B. Randall, MD, MBA

• Lentine, Krista L., Saint Louis University, St Louis, Missouri, United States

Krista L. Lentine, MD, PhD, FASN

• Edwards, John C., Saint Louis University, St Louis, Missouri, United States

John C. Edwards, MD, PhD

• Jain, Ajay, Saint Louis University, St Louis, Missouri, United States

Ajay Jain, MD

• Nazzal, Mustafa, Saint Louis University, St Louis, Missouri, United States

Mustafa Nazzal, MD

• Caliskan, Yasar, Saint Louis University, St Louis, Missouri, United States

Yasar Caliskan, MD

Background

LIMS1 rs893403 homozygous GG genotype in donors is associated with decreased risk of T-cell mediated rejection after kidney transplantation. However, the effects of ischemia and reperfusion injury (IRI) on donor kidney expression profile at single cell level have not been studied in a human kidney model. Using non-utilized human kidneys, we investigated the effects of IRI on the nuclear expression of LIMS1, GCC2, and associated genes in donor kidneys with different genotypes and compared results between ex-vivo normothermic machine perfusion (NMP) model and cold static storage (CS).

Methods

Deceased donor kidneys (n=5) on CS underwent 6-hr NMP (n=5) following CS. Cells at baseline on CS and after 6 hrs of NMP were harvested. Single nucleus transcriptomics of the donor kidneys after 6 hrs of NMP was directly compared with the profile on baseline CS. Specimens sharing the same genotype were subsequently processed together using the droplet-based 10X Genomics approach.

Results

Three kidneys possessed the AG allele (donor rejection risk genotype), while two exhibited the GG allele (donor non-rejection risk genotype) for the LIMS1 gene (rs893403). The study yielded a collection of over 60,000 cells accompanied by 2 billion reads. 13 unique kidney cell types were identified using Seurat (v7.0). SnRNA-seq analyses showed induction of LIMS1 and GCC2 in tubule cells particularly in ischemic donor kidneys with non-rejection risk genotype (GG) after NMP. LIMS1 and GCC2 expression increase is significant in proximal and distal tubule cells after 6 hrs of NMP in non-rejection risk kidneys (GG) (Fig1a, 1b). Transcriptomic response to NMP is different in kidneys with GG and AG genotypes (Fig 1c).

Conclusion

NMP significantly increased the LIMS1 and GCC2 expression on proximal and distal tubule cells with LIMS1 rs893403 GG variant but not on the kidney with AG variant. Donor LIMS1 genotype is important in the transcriptomic response to IRI.