Detection of Plasma DNA of Renal Origin in Kidney Transplant Patients (DART-REIN)

Donor-derived cell-free DNA (dd-cfDNA) has been proposed as a potential diagnostic tool to monitor the rejection status of the kidney transplant. It has been suggested that dd-cfDNA is increasing in the blood of kidney transplant patient presenting a graft rejection. In this project, investigators proposed a different approach to predict and characterize kidney transplant rejection/dysfunction based on the quantification of epigenetic signatures present on the donor-cell-free DNA. In 2018, Moss et al. develops a deconvolution model capable of identifying the tissue origin of circulating DNA by taking advantage of its epigenetic properties. The study confirmed that the cell-free DNA circulating in healthy subjects comes mainly from blood cells and endothelial cells, but not from kidney cells.

In this study, researchers investigate the evolution of blood renal-specific cell-free DNA amount in patient with chronic kidney disease before and after the transplantation surgery by testing a set of renal-specific epigenetic markers. The purpose of this study is to identify the biological noise of "native kidney" on renal-specific cell-free DNA and to compare it with signal coming from "transplanted kidney".

Study Overview

• Status: Recruiting
• Conditions: Acute Kidney Injury; Renal Transplantation
• Intervention / Treatment: Genetic: determination of circulating biomarkers of renal origin circulating biomarkers of renal origin

Detailed Description

At the diagnostic level, routine monitoring of graft functionality after kidney transplantation relies on the use of non-specific markers, such as serum creatinine (allowing estimation of glomerular filtration rate or GFR) and proteinuria. Definitive diagnosis of renal allograft dysfunction still requires invasive allograft biopsy, which remains the gold standard for assessing graft status. The histopathological diagnosis of renal graft dysfunction is based on the Banff classification and makes it possible to examine the immune infiltrate and the cellular lesions of the graft to make a precise diagnosis. However, Renal biopsy has certain limitations: 1/ It is invasive for the patient, with associated complications, mainly hemorrhagic in 1 to 3.5% of cases; 2/ Its effectiveness in predicting early rejection (3 months) post-transplant remains controversial. A lack of patient benefit has been proven by several studies for screening biopsy; 3/ Histological analysis is subject to intra- and inter-observer variations; and 4/ it is an expensive examination due to the medical time required to perform and interpret the biopsy.

The "donor-cell-free DNA (dd-cfDNA)" has been suggested as a diagnostic tool at the service of the graft. Analyzes based on molecular signatures on the circulating DNA of the donor (Single Polymorphism Nucleotide (SNP)) have made it possible to discriminate the circulating cell-free DNA from the transplanted kidney (from the donor) from the circulating cell-free DNA specific to the recipient. Data from several studies suggest that blood dd-cfDNA levels can detect rejection in heart, lung, liver and kidney allografts. First studied by multiplex qPCR and then NGS technologies, the SNPs present on the dd-cfDNA were then quantified by digital PCR techniques.

In this study, investigators proposed a different approach to predict and characterize kidney transplant rejection/dysfunction based on the quantification of epigenetic signatures present on the donor-cell-free DNA. In 2018, Moss et al. develops a deconvolution model capable of identifying the tissue origin of circulating DNA by taking advantage of its epigenetic properties. The study confirmed that the cell-free DNA circulating in healthy subjects comes mainly from blood cells and endothelial cells, but not from kidney cells. In 2023, Loyfer et al. proposed a methylation atlas of more than 200 cells type and suggested that each cell has its own epigenetic signature that can be study in cell-free DNA.

In this study, researchers investigate the evolution of blood renal-specific cell-free DNA amount in patient with chronic kidney disease before and after the transplantation surgery by testing a set of renal-specific epigenetic markers. The purpose of this study is to identify the biological noise of "native kidney" on renal-specific cell-free DNA and to compare it with signal coming from "transplanted kidney". Researchers hypothesize that the biological noise from "native kidney" in chronic kidney diseases is negligible compared to that of the post-transplantation graft.

To investigate this hypothesis, investigators collect blood samples before and after transplant surgery to quantify kidney-specific cell-free DNA markers. They also proposed to quantify cell-free DNA markers of graft perfusion fluid to validate the specificity of renal markers and to study graft tissue damage during organ transport. Each renal-cell-free DNA sample is quantified by a proprietary technologies using a multiplex digital-PCR assay.

• Study Type: Observational
• Enrollment (Estimated): 30

Contacts and Locations

• Study Contact: Name: Geoffroy Dr POULET, PhD; Phone Number: +33 +33 6 67 77 23 94; Email: Geoffroy.poulet@cgenetix.com
• Study Contact Backup: Name: Sarah Dr DROUIN, MD-PhD; Phone Number: +33 +33 1 42 17 71 73; Email: sarah.drouin@aphp.fr

Study Locations

• France
  • Île-de-France Region
    • Paris, Île-de-France Region, France, 75013
      • Recruiting
      • Pitié-Salpêtrière Hospital
      • Contact: Geoffroy POULET, PhD; Phone Number: +33 6 67 77 23 94; Email: geoffroy.poulet@cgenetix.com
      • Contact: Sarah DROUIN, MD, PhD; Phone Number: +33 1 42 17 71 73; Email: sarah.drouin@aphp.fr

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult; Older Adult
• Accepts Healthy Volunteers: No

• Sampling Method: Non-Probability Sample

Study Population

Adult subjects with end-stage renal disease about to receive a kidney transplant from a deceased donor.

Description

Inclusion Criteria:

• Age ≥ 18 years old
• With end-stage renal failure
• Summoned for a kidney transplant at Pitié Salpêtrière Hospital
• Having been informed of the study and not objecting to the study having given free and informed written consent for the genetic analysis
• Benefiting from a social security scheme (excluding AME)

Exclusion Criteria:

Under legal protective measures (curatorship or guardianship, under judicial safeguard).

Study Plan

How is the study designed?

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Amount of renal circulating cell-free DNA	The amount of renal-cell-free DNA (glomerular and tubular markers) will be measured 6 hours before the kidney transplant is performed and 12-24 hours after the kidney transplant by digital multiplex PCR	6 hours before kidney transplantation and 12 to 24 after transplantation surgery

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Estimating the inter-individual variations of the free circulating methylome of renal origin in patients with end-stage chronic insufficiency	The amount of renal-cell-free DNA (glomerular and tubular markers) measured will be compared between individuals.	6 hours before kidney transplantation and 12 to 24 after transplantation surgery
Identify by a method without a priori (methyl seq) specific markers of acute renal injury in terms of epigenetic signature	Comparison of two biomarkers quantification methods (whole genome methyl-Sequencing and multiplex digital-PCR)	6 hours before kidney transplantation and 12 to 24 after transplantation surgery
Study the statistical association between the presence of free circulating methylated sequences of renal origin and the resumption of graft function	Comparison of circulating free methylome of renal origin between the groups of patients with immediate recovery of function and delayed recovery of function (defined on the performance of a dialysis session in the first 7 days and on the reduction ratio serum creatinine between the 1st and 2nd day after the transplant).	7 day after the transplant

Collaborators and Investigators

• Sponsor: Assistance Publique - Hôpitaux de Paris

Study record dates

Study Major Dates

• Study Start (Actual): September 2, 2024
• Primary Completion (Estimated): July 2, 2026
• Study Completion (Estimated): July 9, 2026

Study Registration Dates

• First Submitted: August 30, 2023
• First Submitted That Met QC Criteria: August 30, 2023
• First Posted (Actual): September 7, 2023

Study Record Updates

• Last Update Posted (Estimated): September 24, 2025
• Last Update Submitted That Met QC Criteria: September 23, 2025
• Last Verified: September 1, 2025

More Information

Terms related to this study

• Keywords: biomarkers; Transplantation; Epigenetic; Renal cell-free DNA
• Additional Relevant MeSH Terms: Urogenital Diseases; Male Urogenital Diseases; Kidney Diseases; Urologic Diseases; Female Urogenital Diseases; Female Urogenital Diseases and Pregnancy Complications; Renal Insufficiency; Acute Kidney Injury
• Other Study ID Numbers: APHP230298

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: YES

IPD Plan Description

The procedures carried out with the French data privacy authority (CNIL, Commission nationale de l'informatique et des libertés) do not provide for the transmission of the database, nor do the information and consent documents signed by the patients.

Consultation by the editorial board or interested researchers of individual participant data that underlie the results reported in the article after deidentification may nevertheless be considered, subject to prior determination of the terms and conditions of such consultation and in respect for compliance with the applicable regulations.

• IPD Sharing Time Frame: Beginning 3 months and ending 3 years following article publication. Requests out of these time frame can also be submitted to the sponsor
• IPD Sharing Access Criteria: Researchers who provide a methodologically sound proposal.
• IPD Sharing Supporting Information Type: STUDY_PROTOCOL; ICF

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No
• product manufactured in and exported from the U.S.: No