Trifecta-Kidney Study

Calibrating circulating donor-derived cell-free DNA against molecular biopsy assessments.

Investigators

Philip F Halloran, Jeff Reeve, Soroush Shojai, Sita Gourishankar and the MMDx-Kidney Study Group (ClinicalTrials.gov NCT04239703)

Current screening methods for detecting rejection in kidney transplant patients, such as histology, creatinine levels, and urine protein analysis, have their limitations. To address this, the Alberta Transplant Applied Genomics Centre has developed the Molecular Microscope® Diagnostic System (MMDx)—a groundbreaking molecular assessment tool for evaluating rejection and injury in kidney biopsies based on gene expression.

We are excited to introduce a new screening test: the Natera, Inc. dd-cfDNA Prospera™ test, which focuses on monitoring donor-derived cell-free DNA (dd-cfDNA) in blood. Our study aims to calibrate dd-cfDNA levels detected at the time of indication biopsies against MMDx measurements for different types of rejection, including T-cell mediated rejection (TCMR) and antibody-mediated rejection (ABMR) at various stages, as well as acute kidney injury (AKI) and late injury characterized by atrophy and fibrosis. We also want to determine if Prospera™ blood test can replace kidney biopsy test.

This research involves a comprehensive comparison of blood dd-cfDNA measurements with MMDx results, while also assessing HLA antibodies in the blood. The study is an extension of the INTERCOMEX project (ClinicalTrials.gov ID: NCT01299168) and is currently being conducted across 27 centers in the US, Canada, Australia, and Europe.

ClinicalTrials.gov NCT04239703 »

Trifecta-Kidney Study Publications

  1. Halloran PF, Reeve J, Madill-Thomsen KS, Demko Z, Prewett A, Billings P, et al. The Trifecta Study: Comparing Plasma Levels of Donor-derived Cell-Free DNA with the Molecular Phenotype of Kidney Transplant Biopsies. J Am Soc Nephrol. 2022;33(2):387-400.
  2. Halloran PF, Reeve J, Madill-Thomsen KS, Demko Z, Prewett A, Gauthier P, et al. Antibody-mediated rejection without detectable donor-specific antibody releases donor-derived cell-free DNA: results from the Trifecta study. Transplantation. 2023;107(3):709-719..
  3. Halloran PF, Reeve J, Madill-Thomsen KS, Kaur N, Ahmed E, Cantos C, et al. Combining Donor-derived Cell-free DNA Fraction and Quantity to Detect Kidney Transplant Rejection Using Molecular Diagnoses and Histology as Confirmation. Transplantation. 2022;106(12):2435-42.
  4. Gauthier PT, Madill-Thomsen KS, Demko Z, Prewett A, Gauthier P, Halloran PF, et al. Distinct Molecular Processes Mediate Donor-derived Cell-free DNA Release From Kidney Transplants in Different Disease States. Transplantation. 2024;108(4):898-910.
  5. Halloran PF, Reeve J, Madill-Thomsen KS, Demko Z, Prewett A, Gauthier P, et al. Antibody-mediated rejection without detectable donor-specific antibody releases donor-derived cell-free DNA: results from the Trifecta study. Transplantation. 2023;107(1):709-19.
  6. Madill-Thomsen KS, Hidalgo LG, Demko Z, Gauthier PM, et al. Defining relationships among tests for kidney transplant antibody-mediated rejection. KI Reports 2025; 10(9):3225-38. 
  7. Hidalgo LG, Madill-Thomsen KS, Reeve J, Investigators T-K, Halloran PF. Improving the histologic detection of DSA-negative antibody-mediated rejection in kidney transplants. Am J Transplant. 2025;In press.(https://doi.org/10.1016/j.ajt.2025.08.029).
  8. Hidalgo LG, Madill-Thomsen KS, Reeve J, Investigators T-K, Halloran PF. Improving the histologic detection of DSA-negative antibody-mediated rejection in kidney transplants. Am J Transplant. 2025;In press.(https://doi.org/10.1016/j.ajt.2025.08.029).