Donor-derived Cell-free DNA Shows High Sensitivity for the Diagnosis of Pancreas Graft Rejection in Simultaneous Pancreas-kidney Transplantation

Abstract

Background. Pancreas graft status in simultaneous pancreas-kidney transplant (SPKTx) is currently assessed by nonspecific biochemical markers, typically amylase or lipase. Identifying a noninvasive biomarker with good sensitivity in detecting early pancreas graft rejection could improve SPKTx management. Methods. Here, we developed a pilot study to explore donor-derived cell-free DNA (dd-cfDNA) performance in predicting biopsy-proven acute rejection (P-BPAR) of the pancreas graft in a cohort of 36 SPKTx recipients with biopsy-matched plasma samples. dd-cfDNA was measured using the Prospera test (Natera, Inc.) and reported both as a fraction of the total cfDNA (fraction; %) and as concentration in the recipient’s plasma (quantity; copies/mL). Results. In the absence of P-BPAR, dd-cfDNA was significantly higher in samples collected within the first 45 d after SPKTx compared with those measured afterward (median, 1.00% versus 0.30%; median, 128.2 versus 35.3 cp/mL, respectively with both; P = 0.001). In samples obtained beyond day 45, P-BPAR samples presented a significantly higher dd-cfDNA fraction (0.83 versus 0.30%; P = 0.006) and quantity (81.3 versus 35.3 cp/mL; P = 0.001) than stable samples. Incorporating dd-cfDNA quantity along with dd-cfDNA fraction outperformed dd-cfDNA fraction alone to detect active rejection. Notably, when using a quantity cutoff of 70 cp/mL, dd-cfDNA detected P-BPAR with a sensitivity of 85.7% and a specificity of 93.7%, which was more accurate than current biomarkers (area under curve of 0.89 for dd-cfDNA (cp/ml) compared with 0.74 of lipase and 0.46 for amylase). Conclusions. dd-cfDNA measurement through a simple noninvasive blood test could be incorporated into clinical practice to help inform graft management in SPKTx patients.

Immune Profiling of Peripheral Blood Mononuclear Cells at Pancreas Acute Rejection Episodes in Kidney-Pancreas Transplant Recipients

“…These results correlate with another recent study in which, exploring a gene set to evaluate the expression of ABMR in pancreas graft biopsy, there was no correlation between the presence of DSA and ABMR gene expression (44). Finally, we have recently reported that donor-derived cfDNA was increased in patients with DSA despite the absence of signs of ABMR in graft biopsy (45). Altogether, these studies highlight that histological ABMR may be underdiagnosed in pancreas transplantation, and is important to design larger studies in patients with pancreas ABMR aiming at exploring the molecular and genetic biomarkers, and more in depth functional analysis of peripheral blood mononuclear cells.…”

Donor-derived cell-free DNA as a diagnostic tool in transplantation

“…(Oellerich et al, 2021). Dd-cfDNA detects rejection also in liver (Schuetz et al, 2017;Levitsky et al, 2022), heart (Snyder et al, 2011;De Vlaminck et al, 2014;Khush et al, 2019;Richmond et al, 2020;Agbor-Enoh et al, 2021a;Sorbini et al, 2021), lung (Rosenheck et al, 2022a) and simultaneous pancreas-kidney (Ventura-Aguiar et al, 2022) transplant patients with good accuracy (Table 2). Dd-cfDNA fraction was also significantly elevated for CLAD (De Vlaminck et al, 2015).…”

Droplet digital PCR-based testing for donor-derived cell-free DNA in transplanted patients as noninvasive marker of allograft health: Methodological aspects

“…Analyzing dd-cfDNA has shown great potential as a biomarker for monitoring the allograft health, where an increase of the dd-cfDNA fraction in the recipient's bloodstream will indicate organ damage [2,[8][9][10][11][12]. As a noninvasive marker for organ health, dd-cfDNA testing has been applied for various organs [3,6], including the heart [13][14][15][16][17][18][19][20][21][22], kidney [5,[23][24][25][26][27][28][29], liver [11,29], lung [30][31][32], and pancreas and simultaneously pancreas-kidney transplantation [33].…”