Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Jurak,C.
Comprehensive Liquid Biopsy in Patients after Organ Transplant
- cfDNA/microRNA and immune cell repertoire as markers for tissue damage and organ rejection
Humanmedizin; [Diplomarbeit] Medical University of Graz;2019. pp. 91
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- Autor*innen der Med Uni Graz:
- Betreuer*innen:
- Kashofer Karl
- Altmetrics:
- Abstract:
- Aims: The aim of this pilot-study is to explore the utility of liquid biopsies for monitoring transplant failure. Clinically transplant failure presents as tissue damage of the transplant due to ischemia, reperfusion injury or immunological rejection. Cell death in the transplant is associated with release of graft-derived cell-free DNA and microRNA from damaged tissue. Accordingly, graft-derived cfDNA/cfmicroRNA can possibly be used as a minimally invasive tool for assessment of transplant damage. On the other hand, immunological rejection could be associated with clonal expansion of T-cells targeting the graft. Methods: Patients and sample collections: From August 2018 till August 2020, a prospective study will be performed at the Medical University of Graz. In this study, approximately 200 patients with organ transplantation will be included. Blood samples will be obtained before transplantation, at Day0, Day7-10, Month 28(+/- 7 days), Year1(+/- 1 month) and if the therapy regime changes or a biopsy is needed. Preparation of Nucleic Acids & White-blood cells: Cell-free DNA is extracted with the QIAamp Circulating Nucleic Acid Kit (Qiagen, Hilden, Germany) and allelic profiling is performed using the HID-Ion AmpliSeq Identity Panel. MicroRNA is prepared using the miRNeasy Plasma/Serum Advanced Kit (Qiagen, Hilden, Germany), DNA of mononuclear white blood cells is used for PCR of CDR3 region with specific VDJ-region primers. Results & Conclusions: This pilot study demonstrated the feasibility of comprehensive analysis from RNA and DNA with pilot probes from the established trial. cfDNA can be discriminated up to 0.1% of donor DNA in the recipient´s blood stream. microRNA-profile as well as decrease in TCRB-variability may hint to transplant rejection. We gained approval from the ethics commission at the Medical University of Graz to proceed to a full clinical study. Starting in September 2018 all patients receiving kidney or liver transplantation at LKH Graz will be invited to participate. 3BR aims to collect and analyze consecutive blood samples of 160 Patients over an observational period of 2 years after transplantation. We hope to improve patient health by supporting clinical decisions with information on the emergence and nature of organ rejection.