A Novel Next-Generation Sequencing Approach without Donor-Derived Material for Acute Rejection and Infection Monitoring in Solid Organ Transplantation

Bing Wei,Xin Xu,Dong Xiao,Jibin Zhang,Chunting Zheng,Qing Yang,Mingzhi Ye,Yongping Lin,Danxia Huang,Di Shao,Jian Wang,Chunrong Ju,Wenhua Liang,Qiuhua Deng,Karsten Kristiansen,Liuhong Zeng,Liping Liu,Jianxing He

doi:10.4236/jct.2018.99054

Abstract

Background: Donor-derived cell free DNA (ddcf DNA) has been reported as a universal noninvasive biomarker for rejection monitoring in heart, kidney, liver, and lung transplantation. Current approaches based on next-generation sequencing for quantification of ddcf DNA, although promising, may be restricted by the requirement for donor material, as donor samples may not be available. Methods: We proposed a novel next-generation sequencing approach without donor-derived material and compared the non-donor-derived approach and the donor-derived approach using simulation testing and 69 clinical specimens. We also evaluated the performance for acute rejection and infection monitoring in lung transplantation. Results: The non-donor-derived approach reached similar efficacy as the donor-derived approach with a significant linear correlation of R2 = 0.98. Subsequent validation in clinical specimens demonstrated significant difference between the acute rejection group (4.83% ± 2.11%, mean ± SD) and the non-rejection group (1.61% ± 0.63%, mean ± SD) (P < 0.0001, Welch’s t test). With the cut-off value of 2.999, our approach had 90.48% sensitivity (95% CI, 69.62% - 98.83%), 100% specificity (95% CI, 91.59% - 100%), and AUC 0.9266 (95% CI, 0.8277 - 1.026). The test also had the ability to simultaneously detect infectious agents, especially cytomegalovirus, as compared with the clinical test. Conclusion: The proposed approach without donor-derived material could potentially be used to monitor acute rejection and infection in lung transplantation and may be applied to other types of solid organ transplantation.

Highlights

IntroductionThe lungs require strong abilities for environmental adaptation and immuno-protection against microbial infections
As the respiratory centre, the lungs require strong abilities for environmental adaptation and immuno-protection against microbial infections
The donor fraction was re-calculated from the simulation data abandoning donor genotyping information by the non-donor-derived cfDNA transplant dynamics (NDTD) approach as pre-transplant donor DNA information is likely lacking in long-term and severely affected patients

Summary

Introduction

The lungs require strong abilities for environmental adaptation and immuno-protection against microbial infections. Despite considerable advances and the wide use of immunosuppressant drugs, acute rejection (AR) remains a highly prevalent major complication of transplantations, especially in the first year post-operationally, impacting 50% to 90% of patients [2]. It is recognized as one of the risk factors for the development of bronchiolitis obliterans syndrome, which leads to long-term morbidity and mortality after lung transplantation [3]. Current approaches based on next-generation sequencing for quantification of ddcf DNA, promising, may be restricted by the requirement for donor material, as donor samples may not be available. Subsequent validation in clinical specimens demonstrated significant difference between the acute rejection

Methods

Results

Discussion

Conclusion