Anti-CD47 monoclonal antibody therapy reduces ischemia-reperfusion injury of renal allografts in a porcine model of donation after cardiac death

Abstract

We investigated whether blockade of the CD47 signaling pathway could reduce ischemia-reperfusion injury (IRI) of renal allografts donated after cardiac death (DCD) in a porcine animal model of transplantation. Renal allografts were subjected to 30 minutes of warm ischemia, 3.5 hours of cold ischemia, and then perfused with a humanized anti-CD47 monoclonal antibody (CD47mAb) in the treatment group or HTK solution in the control group (n = 4/group). The animals were euthanized five days after transplantation. At the time of reperfusion, indocyanine green-based in vivo imaging showed that CD47mAb-treated organs had greater and more uniform reperfusion. On post-transplant days 3-5, the treatment group had lower values compared to the control for creatinine and blood urea nitrogen. Histological examination of allograft tissues showed a significant decrease of acute tubular injury in the CD47mAb-treated group compared to control. Compared to the control group, CD47mAb treatment significantly decreased genes expression related to oxidative stress (sod-1, gpx-1, and txn), the inflammatory response (il-2, il-6, inf-g, and tgf-b), as well as reduced protein levels of BAX, Caspase-3, MMP2, and MMP9. These data demonstrate that CD47mAb blockade decreases IRI and subsequent tissue injury in DCD renal allografts in a large animal transplant model.

Pharmacologic targeting of renal ischemia-reperfusion injury using a normothermic machine perfusion platform

“…Interestingly, with the exception of IL-6, there was no reduction in the mRNA expression of TNF-α, IL-1β, CCL2, or CXCL2 in the αCD47Ab-treated mice. A lack of impact on TNF-α mRNA expression in response to CD47 blockade has also been noted elsewhere, indicating this pathway is not directly involved in CD47-mediated cellular injury during IRI 21,48 . The primary focus of this paper was the acute phase of injury secondary to IRI, which manifests as delayed graft function (DGF) in the kidney transplant recipient.…”

“…There was no immunofluorescence evidence that this cold perfusion component of the αCD47Ab dose caused effective binding to its receptor. In contrast, Xu et al showed renal binding after pre-implantation delivery of αCD47Ab to porcine kidneys via a direct renal artery cold flush 48 . However, these authors used a dose that was approximately 50 times greater than that used in this study.…”

Thrombospondin 1 and Its Diverse Roles as a Regulator of Extracellular Matrix in Fibrotic Disease

“…Anti-CD47 antibodies and small molecules that mimic the TSP binding site for bFGF are being tested in clinical trials, in preclinical models, or are under development for various diseases. 169,[178][179][180][181][182] Antagonists of these antiangiogenic TSP1 functions have not been tested with antagonists of TSP1's TGF-β activating function. Combined use of a variety of TSP1 functional antagonists could represent a multipronged approach to treating the complex pathogenesis of fibrosis.…”

Enhanced immunosuppression improves early allograft function in a porcine kidney transplant model of donation after circulatory death

“…20,21 Briefly, the donors and recipients were intramuscularly injected with the analgesic of buprenorphine 0.005-0.1 mg/ kg, and anesthesia was induced with a TKX cocktail (Telazol 4 mg/ kg, ketamine 2 mg/kg, and xylazine 2 mg/kg), and maintained with 2.5%-4% isoflurane in oxygen at a flow of 2.0 L/minute. 20,21 Briefly, the donors and recipients were intramuscularly injected with the analgesic of buprenorphine 0.005-0.1 mg/ kg, and anesthesia was induced with a TKX cocktail (Telazol 4 mg/ kg, ketamine 2 mg/kg, and xylazine 2 mg/kg), and maintained with 2.5%-4% isoflurane in oxygen at a flow of 2.0 L/minute.…”

“…Animals were anesthetized and transplanted as previously described. 20,21 Briefly, the donors and recipients were intramuscularly injected with the analgesic of buprenorphine 0.005-0.1 mg/ kg, and anesthesia was induced with a TKX cocktail (Telazol 4 mg/ kg, ketamine 2 mg/kg, and xylazine 2 mg/kg), and maintained with 2.5%-4% isoflurane in oxygen at a flow of 2.0 L/minute. An abdominal midline approach was used to expose the kidneys and vasculature.…”