Ex Vivo Adenoviral Vector Gene Delivery Results in Decreased Vector-associated Inflammation Pre- and Post–lung Transplantation in the Pig

Abstract

Acellular normothermic ex vivo lung perfusion (EVLP) is a novel method of donor lung preservation for transplantation. As cellular metabolism is preserved during perfusion, it represents a potential platform for effective gene transduction in donor lungs. We hypothesized that vector-associated inflammation would be reduced during ex vivo delivery due to isolation from the host immune system response. We compared ex vivo with in vivo intratracheal delivery of an E1-, E3-deleted adenoviral vector encoding either green fluorescent protein (GFP) or interleukin-10 (IL-10) to porcine lungs. Twelve hours after delivery, the lung was transplanted and the post-transplant function assessed. We identified significant transgene expression by 12 hours in both in vivo and ex vivo delivered groups. Lung function remained excellent in all ex vivo groups after viral vector delivery; however, as expected, lung function decreased in the in vivo delivered adenovirus vector encoding GFP (AdGFP) group with corresponding increases in IL-1 levels. Transplanted lung function was excellent in the ex vivo transduced lungs and inferior lung function was seen in the in vivo group after transplantation. In summary, ex vivo delivery of adenoviral gene therapy to the donor lung is superior to in vivo delivery in that it leads to less vector-associated inflammation and provides superior post-transplant lung function.

Clinical grade allogeneic human mesenchymal stem cells restore alveolar fluid clearance in human lungs rejected for transplantation

“…With EVLP, lungs are perfused and ventilated ex vivo at body temperature to mimic physiological conditions. EVLP can also facilitate delivery of a pharmacological, gene-or cell-based therapy to repair damaged lungs before transplant (23,40). We previously reported that intravenous clinical-grade, cryopreserved allogeneic human mesenchymal stem (stromal) cells (hMSC) were effective in restoring alveolar fluid clearance (AFC) in a human EVLP model of acute lung injury (ALI) induced by endotoxin or Escherichia coli bacteria (22,23).…”

292 Physiologic and Biochemical Profile of Clinically Rejected Lungs on a Normothermic Ex Vivo Lung Perfusion Platform

“…[27] In addition to reparative processes intrinsic to the lungs, Steen's solution, the most commonly used EVLP perfusate, has been specifically designed with a high colloid oncotic pressure in an effort to remove both interstitial and alveolar fluid from damaged lungs. [11] The EVLP platform also allows for physical manipulation with the removal of blood and secretions from the lungs; [10] biochemical manipulation with anti-inflammatory cytokines and vasodilatory agonists; [28][29][30][31] and cellular manipulation with mesenchymal stem cells. [32] In our study, we utilized the novel arginase inhibitor, ABH, in an effort to manipulate human lungs during EVLP.…”

The future of marginal kidney repair in the context of normothermic machine perfusion

“…In porcine models of ex vivo lung perfusion, adenoviral vector delivery has been demonstrated to be safe and effective. This therapy resulted in a decreased inflammatory response with better graft functionality after transplantation 31. In the area of renal research, Brasile et al previously demonstrated the ability to target an exogenous gene to the vascular endothelium of a kidney during NMP in a proof‐of‐concept experiment 32.…”