Ex vivo perfusion-based engraftment of genetically engineered cell sensors into transplantable organs.

Ling-Yee Chin,Biju Parekkadan,Stephen P Schmidt,Heidi Yeh,Cailah Carroll,Danielle M Detelich,Gregory R Wojtkiewicz,Ralph Weissleder,Siavash Raigani,Korkut Uygun,Shannon N Tessier

doi:10.1371/journal.pone.0225222

Abstract

Cellular rejection of liver transplant allografts remains a concern despite immunosuppressant use. Existing transplant biomarkers are often not sensitive enough to detect acute or chronic rejection at an early enough stage to allow successful clinical intervention. We herein developed a cell-based sensor that can potentially be used for monitoring local events following liver transplantation. Utilizing a machine perfusion system as a platform to engraft the cells into a donor liver, we effectively established the biocompatibility of the biosensor cells and confirmed efficient delivery of cells distributed throughout the organ. This work proves an innovative concept of integrating synthetic reporter cells ex vivo into organs as a transplant-within-a-transplant during functional organ preservation with a vision to use cell biosensors as a broad way to monitor and treat tissue transplants.

Highlights

Liver transplantation remains the definitive surgical cure for end-stage liver disease
Once rat fibroblasts were transduced with lentiviral particles, Gaussia luciferase (gLuc) secretion was characterized during a 6-hour in vitro experiment modeled on the planned perfusion with a washout period (Fig 1B)
We explored gLuc release in vitro between typical fibroblast growth media (DMEM), perfusate using a Dulbecco Modified Eagle Medium (DMEM) base, and perfusate using a Williams E base

Summary

Introduction

Liver transplantation remains the definitive surgical cure for end-stage liver disease. The last several decades have seen significant advances in the safety of transplant surgery and outcomes of recipients. Advances in immunosuppression, including the introduction of calcineurin inhibitors and mammalian target of rapamycin (mTOR) inhibitors, have improved rejection rates [1]. Detection of graft rejection has lagged behind these developments. Diagnosis of post-transplant liver dysfunction, including ischemia-reperfusion injury, acute and chronic rejection, and even steatohepatitis, often requires core liver biopsies (percutaneous or endovascular), often in serial fashion. Which is not without risk especially in patients with liver failure. Current serum biomarkers of graft dysfunction are largely limited to nonspecific

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Conclusion